Corazón RV Park
Corazon Line
CORAZON RV PARK
2285 ROLDAN DR
(I-25, EXIT 191 WEST)
BELEN, NEW MEXICO 87002
Authentically New Mexican
EXPLORE LIKE A LOCAL
EXPLORE THE BELEN AREA
New Mexico is a fascinating state. Filled with history, culture, geology, events and weather you just can't beat.
There's lots to see around here.
Where should we start?
The flavors of New Mexico
Plenty of Events throughout the Year
We have mountains and deserts and lakes and rivers for every kind of outdoor activity
If you stay in the Heart of New Mexico, you can access a variety of day trips
Dining Out, Clubs, Events and more.
The Best Places to Visit in New Mexico, USA
The Best Places to Visit in New Mexico, USA New Mexico is a constituent state of the United States of America. It became the 47th state of the union in 1912. New Mexico ranks fifth among the 50 United State states in terms of total area and is bounded by Colorado to the north, Oklahoma and Texas to the east. New Mexico has some of the flattest land as well as some of the most rugged mountains in the country and it was described as The Land of Enchantment. The highest mountain peaks, Wheeler Peak and Truchas Peak are in the Sangre de Cristo Mountains in the north-central part of the state. The Rocky Mountains extend into the north-central part of the state. support : https://paypal.me/pierpaoloromanelli 1.Santa Fe 2.Taos 3.Carlsbad Caverns 4.Albuquerque 5.Chaco Canion National Historic Park 6.White Sands National Monument 7.Bandelier National Monument 8.Kasha-Katuwe Tent Rocks National Monument 9.Gila Cliff Dwellings 10.Very Large Array
Official Best of New Mexico 2013
See some of the very best things New Mexico has to offer in 2013! From Hot Air Balloons to Horseback riding. See some unique New Mexico towns and cities and enjoy some of the greatest stays and vacations in the country, and so much more - see it all here!
"New Mexico True" Statewide Sing-a-Long to "Feels Like Home"
The lyrics talk of the beauty all around New Mexico, so it is only logical that this time around singers from all around the state take turns belting out "Feels Like Home." With the help of the New Mexico Tourism Department, a total of 43 communities and organizations have combined for a unique, humorous, and occasionally-in-tune version of the catchy melody. The original version of "Feels Like Home" is already a music video hit. It has more than 100,000 views since its release in November 2013. (you can see the original here: http://youtu.be/JUhXostcdC4 ) This unusual version is not a lip-sync video. These are the actual voices of several hundred New Mexicans singing about all the things that make the state "feel like home" to them. Tourism Department representatives traveled all around New Mexico throughout a summer capturing each community's voices. The song "Feels Like Home" has served as the catchy background tune for numerous TV and radio ads all across the state. The New Mexico Tourism Department first used the song in its own ads, and several communities have since followed suit. It is the creation of the band Richmond, the musical partnership of Albuquerque natives Nate Boitano and Heath Warren. Nate and Heath first gained a measure of fame as members of the popular Albuquerque band Asper Kourt. They have been friends and musical collaborators since growing up next to each other.
"Feels Like Home (New Mexico True)"
The Albuquerque-based band Richmond unveils the official music video for their song "Feels Like Home (New Mexico True)." *available on iTunes - just search for Richmond, or here's a link: https://itunes.apple.com/us/album/feels-like-home/id624687947?i=624688245 Richmond is Nate Boitano and Heath Warren. The video depicts the summer-long New Mexico adventures of the musical pair and their friends. It shows them exploring many of the state's treasures, and includes scenes from places and adventures they would like to experience in the future. And it's all set to a catchy song the duo wrote about the land they love. "Feels Like Home (New Mexico True)" has served as the catchy background tune for numerous TV and radio ads all across the state. The New Mexico Tourism Department first used the song in its own ads, and several communities have since followed suit. Nate and Heath first gained a measure of fame as members of the popular Albuquerque band Asper Kourt. They have been friends, neighbors and musical partners since their childhood. This is from the duo's Facebook page: "Fate, destiny, or guided wind? Call it what you want. But this songwriting duo was born to make music together. Richmond is an acoustic brotherhood made up of two lifelong best friends and next door neighbors. Growing up playing sports, catching lizards, and mastering their musical craft, Nate Boitano and Heath Warren are leaving their marks on the music scene with a whole new approach." #NewMexicoTrue
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Warshal: A Jewish take on Martin Luther King Day
By Rabbi Bruce Warshal
Jan 23, 2017 | 10:26 AM
Rabbi Bruce Warshal (Lindsay Moore / Courtesy)
Earlier this week our nation celebrated the Martin Luther King holiday. For some people it meant only that they had a long weekend vacation, but truly it is one of our most significant holidays. Yes, we honor the man, but more important we celebrate what he stood for – an America committed to equality, a statement that we reject prejudice against all people, a concept that is being challenged this very moment in our country.
I have a personal relationship with this holiday that I share with you, but most important, I want to share what I have learned from my experiences in the civil rights movement. In 1961 when I was a law student in New Haven, Connecticut, my late wife, Lynne, and I traveled south, crossing the Mason-Dixon Line into the heart of the segregated State of Maryland. There we joined CORE (Congress of Racial Equality), a cutting edge, non-violent civil rights organization that was a couple of degrees to the left of Martin Luther King. We participated in sit-ins, and only at one time were we actually in fear for our lives. But, thank God, Maryland was not Mississippi or Alabama, and we were rescued by the Maryland State Troopers, rather than being attacked by racist police in the Deep South.
Upon graduation I practiced law in Cleveland, Ohio and became vice-president of the local CORE organization. In August of 1963 as a group we traveled to Washington, D.C. to be part of what became the historic March on Washington. I feel humbled and appreciative that I was able to be a witness to the great "I have a Dream speech" that is now considered by historians to be one of the most important American speeches, along with the Gettysburg Address and Washington's Farewell Address in 1796.
Cleveland was a segregated community equal to many in the south – not segregated by law, but by ingrained personal prejudice. As we worked to integrate motels and restaurants and even St. Luke hospital, where I am proud to report my wife was captain of the picket line, we were labeled communists by the daily newspaper, the Cleveland Plain Dealer. We were also monitored by the Anti-Subversive Unit of the Cleveland Police Department that reported directly to the FBI, headed by the infamous J. Edgar Hoover, who believed that Dr. King was also a communist. When we attended CORE meetings they would note our license plates and snap pictures. Our phones were tapped and our names appeared in the local press.
My wife and I played a small part in a great movement in this country and I received more from the experience than the movement received from my participation. Let me share with you what I learned:
[More from the Jewish Journal] Stories of freedom need to be told »
First, segregation was not exclusively a southern problem. We northerners could disavow the "original sin" of slavery that was actually written into our Constitution, feeling superior to our southern brethren. But slavery was legal in the northern colonies as well, and slavery was not abolished in New York State until 1827. The virus of prejudice that replaced actual slavery was just as common throughout America – in the north, in the heartlands, in the west, as in the south. "They" were not the only guilty ones. "We" were all guilty as well, and today in our own communities we are still susceptible to this virus. Vigilance is still required.
Second, I learned that it is most important to speak truth to power, not to remain silent even when the "respectable" leaders of the community brand you a rebel, a trouble-maker, or worse, an unpatriotic subversive. Rabbi Joachim Prinz, originally of Berlin, was a rabbi under the Nazis until he was expelled in 1939. He became the rabbi of the Reform synagogue in Newark, New Jersey and an outstanding advocate for civil rights. He was one of the ten organizers of the March on Washington, and as such he delivered a six minute speech immediately preceding Dr. Martin Luther King. Today Rabbi Prinz is remembered for this passage from his speech: "When I was the rabbi of the Jewish community in Berlin under the Hitler regime, I learned many things. The most important thing that I learned under those tragic circumstances was that bigotry and hatred are not the most urgent problem. The most urgent, the most disgraceful, the most shameful and the most tragic problem is silence."
Third, I learned to overcome fear. When Senator Strom Thurmond of South Carolina, then a committed segregationist, spoke in Cleveland, we in CORE picketed his appearance. That afternoon my wife received a telephone call saying that if her husband was part of the picket-line she would be a widow the following morning. Of course I showed up, because we cannot be paralyzed by fear. If fear predominates, tikun olam (the repair of this world) will never be accomplished.
I am writing this in a Jewish newspaper, so it is appropriate to talk about our Jewish contribution to the civil rights battle. With pride I report that Jews have always been in the forefront of progressive social changes. This was certainly true of the civil rights movement. 50% of the whites involved were Jewish, although we were only two and one-half percent of the population. It is also believed that at least 50% of the money that funded the movement came from Jewish sources.
[More from the Jewish Journal] Help your loved ones fight fraud »
As a young man, I was inspired by our rabbinic leadership. Rabbi Arthur Lelyveld of Cleveland suffered a concussion when he was beaten with a tire iron by segregationists in Mississippi. Rabbi Yoachim Prinz walked picket-lines in cities throughout this country as well as participating in the March on Washington. Rabbi Jacob Rothschild fought segregation in the heart of Georgia, leading to the famous 1958 bombing of his congregation (The Temple) in Atlanta. Rabbi Abraham Joshua Heschel, a professor at the Jewish Theological Seminary, marched with Martin Luther King. These giants of rabbinic leadership, along with scores of other rabbis across this country, inspired me to leave the practice of law and to enter the rabbinate.
Now a word about the Rev. Dr. Martin Luther King: His life, his face, his name have become symbols of the American quest for justice, the very meaning of this holiday. But he was in real life more than a symbol. He was a flawed human being, as we all are, but a great man worthy of emulation, for two primary reasons:
First, he was not merely a brilliant writer and orator, capable of inspiring a nation. He was a man of action. In today's lingo, he not only talked the talk, but he walked the walk. He put his body on the line, and we are aware of his tragic end. Not only his words, but his actions inspire us today to continue the work of tikun olam.
Second, and this is a most important point, he did not limit his energy to the plight of his own people. He ignored advice to remain focused on segregation when he spoke out against the evil of the Vietnam War in his famous 1967 speech at the Riverside Church in New York City. He was in Memphis to support workers' rights in a sanitation strike, an issue totally distinct from segregation, when he was assassinated.
Stories of freedom need to be told
Help your loved ones fight fraud
First moon landing was important for Jewish community, too | Opinion
His example has affected my life. I learned that as we become better Jews, we become better human beings. As we become more Jewish, we also become more interested in all humanity. Reality is reflected in this paradox. I never felt more Jewish than when I played my small part in the civil rights movement.
This lesson also applies to the very Jewish assertion of "Never Again." It should not only relate to the fate of us Jews, but to the concept that never again should a genocide afflict humanity as a whole. Dr. Martin Luther King taught us how to go from the particular to the universal, from caring for one's own people, to caring for all humanity. Our rabbis taught us that same principle. Almost 2,000 years ago the Talmud stated that Passover celebrates not merely the history of Jews going from slavery to freedom but the concept of all peoples emerging into freedom.
It is this, the true meaning of Martin Luther King Day, an iteration of our ancient Jewish values, that was celebrated earlier this week.
Rabbi Warshal is the publisher emeritus of the Jewish Journal and the author of "Provocative Columns: A Liberal Rabbi Reflects on Beliefs, Israel and American Politics" available at Amazon and Barnes & Noble. He can be reached at brucewarshal@gmail.com


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New service aims to nix bad mortgage leads
LeadPolice plans to start 'policing' real estate leads, too
by Matt Carter |
A group of marketing professionals has formed a company they say is making it easier for mortgage brokers to buy leads from trusted sources.
While other companies help connect lead buyers with leads, they typically charge lead sellers to advertise on their sites or take a cut of their profits.
While other companies help connect lead buyers with leads, they typically charge lead sellers to advertise on their sites or take a cut of their profits. LeadPolice.com turns that business model on its head, letting sellers who meet the company's standards market themselves on the site for free, while charging buyers to use the service.
Because LeadPolice doesn't depend on sellers for revenue, buyers can be assured the company has their best interests in mind, said LeadPolice.com Vice President Melissa Rosno. LeadPolice.com owes its allegiance to buyers, who share information about the quality of leads they obtain from approved sellers. Companies that fail to deliver are barred from the site.
"A lot of Web sites out there have information on lead-generation companies, but for the most part they are charging money on advertising," Rosno said. Because ending a relationship with a seller can hurt those companies' bottom lines, she said, "It's hard for them to remain unbiased."
"To remain a truly unbiased third party, we don't think there can be a revenue exchange between the lead sellers and the intermediary," Rosno said. "We can honestly say that, unlike other companies that take a percentage of revenue, we have no motive to keep (sellers of bad leads) in our database."
Lead buyers pay $30 a month or $300 per year for membership, a cost they can easily recoup through discounts sellers offer to members and higher-quality leads, Rosno said. Because sellers are getting free advertising on the site, LeadPolice.com encourages them to offer discounts to its members.
With mortgage leads often selling for $50 or more these days, if sellers offer a 10 percent discount to LeadPolice members, that "adds up pretty quickly," Rosno said. "The savings can pay for membership."
The site also offers dispute resolution, and aims to be an educational forum where buyers research authorized lead sellers and learn from industry experts.
LeadPolice provides information on companies such as how many years they have been in business, the average rate of return on their leads, and the number of leads they generate — 25 "data points" in all, Rosno said.
"I'm very excited about the ability to compare companies side by side," Rosno said. "Obviously the more informed you are the better decision you can make. Never before has this information been put together in one place."
LeadPolice.com launched this summer with mortgage lead originators, and plans to offer real estate leads by the end of the year, Rosno said.
"We chose to launch with mortgage because there are more mortgage lead-generation companies than any other category," Rosno said. "Mortgage brokers, especially smaller ones, are more likely to rely on purchased leads than real estate agents."
While real estate agents are likely to specialize in a local territory, mortgage brokers can lend statewide or even nationwide.
LeadPolice has signed up "a lot of the big players" in the mortgage lead-generation business, including Prospect Zone and LoanWeb, and now has about 40 authorized lead sellers, Rosno said.
Some other Web sites that offer to match buyers of mortgage leads with multiple sellers include ROOT Exchange and LeadPoint.
ROOT Exchange and LeadPoint are modeled after commodities exchanges, with sellers competing for buyers who bid for leads at auction.
Both companies say they screen sellers and monitor the quality of the leads sold on their sites, and will bar companies that aren't providing good ones.
"One hundred percent of the leads coming through our system are getting feedback," said LeadPoint chief marketing officer Michael Rosenberg. "It's not just a subjective, or anecdotal rating" of a company, but customer evaluations of the quality of each bundle of leads sold on the site, he said.
"When we find out a seller is providing poor-quality leads, we kick (them) out of the marketplace without the buyers even finding out," Rosenberg said. "We're doing all the work for them."
Although LeadPoint is an exchange for many types of leads — nearly 1 million have been bought and sold on the site this year alone — mortgage leads helped the company set a single-day record of 25,000 trades on Oct. 16.
More than 350 lenders purchased leads generated by a LeadPoint seller that placed an ad on the MSN homepage offering home equity and refinance loans. LeadPoint estimates the leads will generate over $100 million in loan originations.
LeadPoint vice president of operations Donny Hall said a potential shortcoming of LeadPolice is that it only helps buyers find a lead seller — it doesn't help facilitate the sale of the leads.
"If you're a buyer going into the LeadPolice directory, to benefit from dealing with five sellers or 100 sellers, you have to develop relationships with each one, and manage them using your own methodology," Hall said. "That's cumbersome."
LeadPolice's Rosno said there are many software applications that help lead buyers manage purchases from multiple sellers.


Jim Harbaugh believes players deserve a cut of "massive TV deals"
"Yeah, I think players should receive a revenue share."
NCAABy Kevin Harrish on 09/05/2022
Before the start of the college football season, theBig Ten signed a series of massive new media rights deals that will generate an average of at least $1.1 billion per year for the conference. While the athletes won't see any of that money directly, a few prominent figures in the conference are calling for that to change – and you can add Michigan head coach Jim Harbaugh to that chorus.
During his weekly media availability on Monday, a reporter asked Harbaugh whether he believed players should get a cut of the multi-billion-dollar television contracts. The Michigan coach offered a blunt and straightforward response.
"I think you know exactly where I stand on that. Well-documented that I do believe the players should receive a revenue share from the massive TV deals that have been worked out," Harbaugh said, according to Wolverines Wire. "As I said back in the summer, a large piece should come the ones who are negotiating those TV deals. It's seven-point some billion? It's a lot. It's a lot of money. Yeah, I think players should receive a revenue share."
Harbaugh is certainly not alone in this stance. Ohio State quarterback C.J. Stroud said last month that he believes revenue should be shared with athletes. But perhaps most notably, Big Ten Conference commissioner Kevin Warren has said he's open to the idea of revenue sharing.
At Big Ten Media Days in July, Warren said he has begun conversations about revenue-sharing with a newly-formed student-athlete advisory and advocacy committee.
"I've already started some dialogue with our student-athletes," Warren said in July. "I want to be a great listener to figure out what is important to them. It's so easy to talk about money and share money, but what does that really mean? I want to make sure that I listen and learn to be able to have big ears and a small mouth to truly understand what's important to them."
During the Aug. 23 edition of Real Sports With Bryant Gumbel, Warren reiterated that stance, telling Gumbel he could foresee the conference or individual teams directly paying athletes one day.
[Wolverines Wire]
Big TenJim Harbaughmedia rightsrevenue sharing
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Chris Russo rules out going to war with WFAN if Michael Kay retires
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Trying to find the world's favourite Stephen King works.
Thread: Trying to find the world's favourite Stephen King works.
Matthew Craig
Junior Member Lurker
Glenavy, Northern Ireland
I've been a long-time Cemetery Dance customer, and have been visiting the forums since they started, but only now registering because I have something to say. Some of you will probably recognise me from days past at Shocklines and the like.
Anyway...I'm currently in the process of celebrating the 40th anniversary of the publication of Carrie on my blog, and as part of those celebrations, I'm trying to find out what are the world's favourite Stephen King works. To this end, I have created a survey, and would love to hear everyone's thoughts.
I'd be very grateful if you could spread the word as much as possible.
Matt Craig
Only the old stuff brother, only the old stuff LOL! Done and done good luck with your project.
There's a definite pattern emerging, Mike, and it seems you're not alone :-)
Teriw
Senior Member 2nd Rubber Room Confinement
I started to do your survey and got frustrated with the way it is set up like having to drag from the bottom to the top waiting for the page to scroll back up and the fact that I would have had to do the green mile and the dark tower separately.
subie09lega
Senior Member Receiving Daily Medication
I agree that the Green Mile selection should be one choice but that's only because I read it as one book and don't distinguish between the six as separate. The DT books though are very discernible as separate selections.
Thanks for the feedback folks.
Teri > As a result of your comments, I have moved the survey to a new host, and completely changed how it works. Hopefully it's a lot more user-friendly on both computers and tablets (and maybe even phones!). Please do give it another try (link below).
subie09lega > The Green Mile is split into its component parts (because that's how I originally read them), but there is also a "Green Mile: The Complete Serial Novel" option for those who wish to go for the full story.
Everyone else: if you've already completed the survey, there's no need to do so again. If you haven't, I promise it's much easier now. You can find the link to the survey at the original post on my blog. I appreciate you all taking the time, and hope that you'll spread the word.
srboone
Senior Member Ok, I really can't come up with anymore of these stupid things...
I wish The Stand was separated into two entries: one for the 1978 version and one for the 1990 version. But I filled out the survey in its original form and had no problem with it.
"I'm a vegan. "
---Kirby Bliss Blanton , The Green Inferno (2013)
cycloneed20
Junior Member Part-timer
Biloxi, Mississippi, United States
A survey about Stephen King books that don't allow me to type in "The Shining" and "The Stand" is a worthless survey.
In order to alphabetise the entries, I've gone for the standard format of "Shining, The", etc. So, if you type "Shining", it'll find the correct title.
Sorry for the confusion, and thanks for looking!


We Love DC
Your Life Beyond The Capitol
Food Truck Tracker
9:00 am08 Nov 2011
David Wax Museum- 930 Club by sightlyworn
As a guy who has photographed many musicians, I know the challenge in capturing the intensity and energy of a live show. Slightlyworn nailed it with this photo. The motion blur is what really makes this photo, because you can just see the guitarist moving and howling into the mic, pouring his heart into every verse. It's a strong composition; he didn't focus too closely on the musician, which left plenty of room for the purple and blue divide caused by the stage lights, which is one of the most important features of the photo. Adding a second singer to the frame and including part of the drum kit definitely frames the context of a musical machine working together. Well done, sightlyworn.
Remember to contribute your best shots of DC to our flickr pool. You may just get noticed for our featured photo or weekend flashback.
Michael T. Ruhl
Michael came to DC in 2009 via Albuquerque, NM. In his spare time, Michael is a photographer, an audiophile, and a guitar aficionado. His humor is drier than the desert and he often finds himself neck deep in awkward situations. Visit his website.
This entry was posted in: Featured Photo
Tagged 9:30 Club, concert, david was museum, Featured Photo, michaeltruhl, musician, slightlyworn
One thought on "Featured Photo"
Aaron McGlauchlen@Sell My Car on November 9, 2011 at 12:24 am said:
This picture reminds me of the one LIVE concert of Bon Jovi that i had seen in 2003, I was fortunate to stand very close on the ramp and wow what energy Jon Bon Jovi, Richie Sambora, David Bryan, Alec John Such and Tico Torres had! I really liked this picture!
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G02B 21/14(2006.01)
G01N 15/02(2006.01)
B82Y 35/00(2011.01)
MA MD
Fundació Institut de Ciències Fotòniques
08860 Castelldefels (Barcelona) (ES)
Institució Catalana De Recerca I Estudis Avançats (ICREA)
08010 Barcelona (ES)
LIEBEL, Matz
08860 CASTELLDEFELS (ES)
HUGALL, James Tom
VAN HULST, Niek F.
(74) Representative: Oficina Ponti, SLP
C. de Consell de Cent 322
(54) A COMMON-PATH INTERFEROMETRIC SCATTERING IMAGING SYSTEM AND A METHOD OF USING COMMON-PATH INTERFEROMETRIC SCATTERING IMAGING TO DETECT AN OBJECT
(57) The present invention relates to a common-path interferometric scattering imaging system and a method of using such a system, where the system comprises:
- illuminating means for emitting an illumination beam (L0);
- light collecting means for collecting through a common collection optical path:
- a scattered beam (Ls) provided by the light scattering on an object (T) of the illumination beam (L0); and
- a reference beam (Lr) provided by the reflection on or transmission through an interface (I) of the illumination beam (L0);
- image sensing means (D) for receiving and sensing the collected scattered (Ls) and reference (Lr) beams interfering thereon as an interferometric light signal;
- attenuation means arranged in the common collection optical path for attenuating the reference beam (Lr) before it arrives at the image sensing means (D); and
- processing means (P) to process data corresponding to the interferometric light signal.
[0001] The present invention relates to a common-path interferometric scattering imaging system and to a method of using common-path interferometric scattering imaging to detect an object, providing an enhanced detection sensitivity.
[0002] There are many different standard scattering imaging systems which use techniques known in the art which constitute only technological background to the present invention, some of which will be briefly described below.
[0003] Phase contrast microscopy (PCM) was introduced from the 1950s, and is a technique which uses phase masks for shifting the phase of scattered light relative to transmitted light to enhance contrast in biological samples due to the large phase shifts introduced by the biological material, such as cells, being studied. As the object of interest shrinks in-size, this technique quickly becomes irrelevant as the phase shift introduced by the biological material becomes small and the scattering signal (which scales as D6, where D is the dimension of interest) is rapidly dwarfed by the background light. For modern studies this has meant that PCM is an abandoned technique when trying to look at sub-cellular elements such as individual proteins, where currently fluorescence microscopy is the standard technique.
[0004] Dark-field microscopy is another well-established and documented microscopy technique with literature and patents dating from the early 20th century. Like PCM it relies on detecting the scattering signal from a sample. It uses a dark-field mask which completely blocks any background light, allowing only the scattered light to be detected. Again this means that for small particles, due to the unfavourable scaling of scattering signal, the technique becomes very hard to implement due to the low number of photon counts to background noise and thus is not used.
[0005] Common-path interferometric scattering imaging systems comprising the features included in the preamble clause of claim 1 of the present invention are known in the art. These systems are usually called iSCAT, and constitute a modern take on phase contrast microscopy (see Lindfors et al. PRL, 93, 3 (2004) - Modern method describing reflection based iSCAT technique, and Piliarik et al. Nature Communications, 5,4495 (2014). These systems generally use a coherent light source (or at least a light source emitting extremely short coherence length light) to generate a reference beam from reflection of the glass/water interface of the coverslip within the focal volume of a microscope objective. The light also generates scattering from particles in the sample. Both the scattered and reference beam are then collected by the objective and imaged onto a camera where they interfere. The common-path nature of this interferometric setup, with reference and scattered beams generated at practically the same position ensures a phase-locked relationship between the two and great stability.
[0006] Common-path interferometric microscopic techniques rely on enhancing small signals using a reference beam to boost a small signal term by interfering the two beams. This provides so-called noiseless gain. The reference and the signal or scattering beam originate from the same light source as coherence between the two signals is essential. In general the intensity on a detector caused by two interfering beams can be described as:
[0007] Where r is the relative reference beam amplitude, s is the relative signal beam amplitude and θ the phase difference between the reference beam and signal or scattering beam.
[0008] As the signal of interest is small (i.e. when the scattering beam is weak, generally due to the object being small), the second term (s2) vanishes compared to the other two terms. The key term of interest is the interference term (2rs cos θ) which includes the signal of interest. All existing interferometric microscopy techniques solely try to maximise this interference term.
[0009] Indeed, existing techniques usually optimise the phase difference θ between the beams to maximise the interference term and, by increasing the power of the illumination light beam, increase the power of the reference beam [essentially the system gain] to increase the signal, up to the saturation point of their detectors.
[0010] By comparing successive images with and without the object of interest, the background reference beam term (r2), which in a good system remains constant, can be removed leaving only the interference term. In the perfect system, this means that the only source of noise will come from the photon or shot noise caused by the total light falling on the detector. Since this noise scales with
where n is the number of photons on the detector, the more photons the camera can record the better the signal-to-noise.
[0011] The fact that the reference beam is proportional to the gain of the system and that the more photons collected the better the signal-to-noise, has led to a focus on finding detectors with large photon [electron full well] capacity to maximise the signal.
[0012] Therefore, existing iSCAT systems, i.e. common-path interferometric scattering imaging systems, teach away from carrying out a variation in the power of the reference beam other than an increasing thereof, in order to maximize the interference term (2rs cos θ) and thus enhance the signal-to-noise ratio. This reference beam power increase approach has, among others, the drawback associated to the need of using expensive detectors (with large photon capacity).
[0013] Hence, it can be stated that iSCAT combined with a well-stabilized laser light source and expensive cameras with large full well capacity and low noise, has allowed the detection and tracking of small particles, despite the small scattering signal on top of a large background, but that the cost to implement this technique as well as the skill required, however, has become prohibitively expensive and prevents its use on a large scale.
[0014] It is, therefore, necessary to provide an alternative to the state of the art which covers the gaps found therein, by providing a common-path interferometric scattering imaging system including an alternative mechanism to enhance detection sensitivity which does not possess the above mentioned drawbacks of the existing iSCAT systems.
[0015] To that end, the present invention relates, in a first aspect, to a common-path interferometric scattering imaging system comprising, in a known manner:
illuminating means comprising a light source configured and arranged for emitting an illumination beam along an illumination optical path including at least two different phases of matter;
light collecting means configured and arranged for simultaneously at least partially collecting through a common collection optical path:
a scattered beam provided by the light scattering by an object of a portion of said illumination beam, wherein said object is placed in at least one of said two different phases of matter; and
a reference beam provided by the reflection on or transmission through an interface of another portion of said illumination beam, wherein said interface is a surface forming a common boundary among said two different phases of matter;
image sensing means configured and arranged for receiving and sensing the collected scattered and reference beams interfering thereon as an interferometric light signal; and
processing means connected to said image sensing means to receive data corresponding to said interferometric light signal, and configured to process said received data to at least detect said object, and, optionally, also to track the object.
[0016] In contrast to the known common-path interferometric scattering imaging systems, the one of the first aspect of the present invention comprises, in a characterizing manner, attenuation means arranged in the above mentioned common collection optical path for attenuating said reference beam before it arrives at the image sensing means.
[0017] The processing means implements an algorithm to process the received data according to the following equation:
where r is the normalised reference beam amplitude, s is the normalised scattering beam amplitude, θ is the phase difference between the reference and scattering beams, α is the attenuation amplitude defined as the reciprocal transmission amplitude, Itotal is the total intensity of the light on the image sensing means caused by the two interfering reference and scattering beams, and I0 is an initial light intensity on the image sensing means, wherein said attenuation means have a degree of attenuation for said reference beam calculated with the purpose of maximizing the term
cos θ with respect to the term
of the above equation to enhance detection sensitivity.
[0018] In other words, the system of the present invention do the exact opposite of what existing iSCAT systems have logically been aiming for: to attenuate the reference beam.
[0019] This is so because, in contrast to the prior art iSCAT systems, instead of focusing purely on signal-to-noise, the system of the first aspect of the present invention highlights an alternative mechanism to enhance detection sensitivity by enhancing the signal-to-background, i.e.
cos θ relative to
By attenuating the reference beam after it the signal beam, i.e. the scattering beam, has been created (i.e. after the scattering event and in collection), the interference term is maximised, as the interference term scales linearly with reference beam amplitude relative to the rapidly decreasing reference beam background which scales quadratically. This increases the signal-to-background and compensates exactly for the loss in signal-to-noise due to increase in shot noise due to lower number of photons collected. Not only does this allow (far more economical) detectors with smaller electron full well capacities, but for larger full-well-capacity cameras, the initial light (i.e., illumination beam) incident on the object can be further increased to increase scattering intensity and increase detection sensitivity, i.e. maximising both signal-to-noise and signal-to-background.
[0020] The attenuation means of the system of the present invention does not fully attenuate the reference beam (as in DFM), and unlike in PCM it does not introduce any significant phase delay between the signals. In fact it works to attenuate the reference beam in amplitude relative to the scattering beam to maximise contrast in an interference setup. Therefore it is conceptually very different from both of the above techniques, neither is it a combination of the above two techniques, but a new form of interference scattering microscopy suitable for detecting small particles of increasing importance in biological sciences as well as many other industrial processes such as nanotechnology.
[0021] For an embodiment, the attenuation means comprises a partially transmissive mask having a semi-transmissive region arranged in a corresponding region of the common collection path through which the reference beam travels, such that the reference beam is attenuated on transmission before reaching the image sensing means.
[0022] For an alternative embodiment, the attenuation means comprises a partially reflective mask having a semi-reflective region arranged in a corresponding region of the common collection path through which the reference beam travels, such that the reference beam is attenuated on reflection before reaching the image sensing means.
[0023] According to specific implementations of any of the above two embodiments, the semi-transmissive or semi-reflective region of the mask is a first region of said partially transmissive or partially reflective mask, the mask comprising a second region arranged in a corresponding region of the common collection path through which part of the scattering beam travels, such that said part of the scattering beam traverses said second region or is reflected thereon thereby before reaching the image sensing means, by transmission or by reflection, wherein said first and said second regions have different transmissive or reflective properties and said partially transmissive or partially reflective mask maintains the coherence relationship between the reference and scattered beams.
[0024] Preferably, said second region is a fully or substantially fully transmissive or reflective region, although for less preferred embodiments the second region can also have some degree of light attenuation.
[0025] According to a preferred embodiment, the first region of the partially transmissive mask has a circular or cylindrical shape and said second region has an annular or tubular shape with an inner diameter larger than the diameter of said first region and being arranged concentrically with respect thereto.
[0026] Other types of optical attenuators, which are not constituted by a mask, are also encompassed for other less preferred embodiments of the system of the first aspect of the present invention.
[0027] The illumination optical path and the common collection path are configured and arranged such that the reference and scattered beams are generated at such closer positions that ensure a phase-locked relationship between the reference and scattered beams, so that there is no need for varying or adjusting the phase of any of said beams. Hence, the system is absent of any phase varying mechanism for said reference and scattered beams as there is no need for phase adjusting.
[0028] According to an embodiment, the system of the first aspect of the invention comprises a coverslip for the object, wherein the above mentioned interface is the common boundary surface among said coverslip and a medium into which said object is placed, the material of which said coverslip is made being non-index matched with said medium.
[0029] According to a first implementation of the system of the first aspect of the invention, called herein as reflective mode, the light collecting means are configured and arranged for collecting said reference beam provided by the reflection on said interface of said another portion of the illumination beam, wherein the system comprises an objective lens which forms part of both the illuminating means and the light collecting means and which is configured and arranged in both the illumination and the collection optical paths to, respectively:
focus the illumination beam into the back-focal plane of said objective lens to produce illumination out of the front aperture of the objective lens, such that a portion thereof will be reflected by the interface generating the reference beam and the rest will pass through the interface up to the object generating the scattering beam; and
receive and at least partially collect both the reference beam and the scattering beam.
[0030] Preferably, the back-focal plane of the objective lens is focused with the illumination beam to produce plane-illumination out of the front aperture thereof, although, for other embodiments, any illumination can be produced as long as it maintains spatial coherence over the time of measurement.
[0031] For an embodiment of said reflective mode implementation, the objective lens is configured and arranged such that the reference beam exits the objective lens as a diverging beam from the centre of the objective lens, when it entered as a plane wave, and passes through or is reflected on the first region of the partially transmissive or partially reflective mask, and the scattering beam leaves the objective lens as a plane wave across a full back-aperture of the objective lens, when it entered as a spherical wave, and passes through or is reflected on both the first and the second regions of the partially transmissive or partially reflective mask.
[0032] For an embodiment, the above mentioned first region of the partially transmissive or partially reflective mask is also placed in the illumination optical path and is configured and arranged to reflect the illumination beam coming from the light source towards the back-focal plane of the objective lens. Other alternative optical mechanisms (prisms, mirrors, etc.) and arrangements for directing the illumination beam towards the objective lens are also encompassed by the system of the first aspect of the invention.
[0033] According to a second implementation of the system of the first aspect of the invention, called herein as transmissive mode, the light collecting means are configured and arranged for collecting said reference beam provided by the transmission through said interface of said another portion of the illumination beam, wherein:
the illuminating means comprises an illumination objective lens configured and arranged to focus the illumination beam into the back-focal plane of said illumination objective lens to produce plane-illumination out of the front aperture of the illumination objective lens, such that a portion thereof will be scattered by the object generating the scattering beam which will be transmitted through the interface, and another portion will be directly transmitted through the interface generating the reference beam; and
the light collecting means comprise a collection objective lens configured and arranged to receive and at least partially collect both the reference beam and the scattering beam.
[0034] For an embodiment of said transmissive mode implementation, the collection objective lens is configured and arranged such that the reference beam exits the collection objective lens as a diverging beam from the centre of the collection objective lens, when it entered as a plane wave, and passes through or is reflected on the first region of the partially transmissive or partially reflective mask, and the scattering beam leaves the collection objective lens as a plane wave across a full back-aperture of the collection objective lens, when it entered as a spherical wave, and passes through or is reflected on both the first and the second regions of the partially transmissive or partially reflective mask.
[0035] Although preferably the attenuation degree provided by the attenuation means has a fixed value, for other embodiments it is adjustable manually or automatically based on the specific use needed at any moment and on parameters associated thereto, such as the size of the object(s) to be detected (and generally tracked), the environmental conditions (light, temperature, etc.), etc., in order to selectively optimising intensity of the reference beam relative to scattered beam to optimise interference contrast on the image sensing means.
[0036] An implementation for providing such adjusting of the attenuation degree of the attenuation means comprises, for an embodiment, a mask having adjustable transmissive or reflective properties and a control system connected to said mask to provide the latter with a control signal (such as an electrical signal) which makes it vary its transmissive or reflective properties as desired, the control signal being created whether in response to manual input of data by a user or automatically based on the sensing of such data by corresponding sensors included in the system.
[0037] A second aspect of the invention relates to a method of using common-path interferometric scattering imaging to detect an object, comprising, in a known manner:
emitting an illumination beam along an illumination optical path including at least two different phases of matter;
simultaneously at least partially collecting through a common collection optical path:
a scattered beam provided by the light scattering on an object of a portion of said illumination beam, wherein said object is placed in at least one of said two different phases of matter; and
receiving and sensing, on image sensing means, the collected scattered and reference beams interfering thereon as an interferometric light signal; and
receiving and processing data corresponding to said interferometric light signal to at least detect said object, and, optionally, also to track the object.
[0038] In contrast to the known methods of using common-path interferometric scattering imaging to detect an object, the one of the second aspect of the present invention comprises, in a characteristic manner, attenuating the reference beam in the common collection optical path before it arrives at said image sensing means.
[0039] Embodiments of the method of the second aspect of the invention comprise the use of the system of the first aspect for all the embodiments thereof describe above.
[0040] For the system and method of the present invention an extremely short coherence illumination light is desirable, whether by using a coherent light source or a light source not considered "coherent" but which generates light with a coherence short enough and with enough power to allowing the above described light interference to occur. In other words, suitable light sources can be lasers or even LEDs.
[0041] In the present document, the term "beam" has been used for referring to light. Alternatively, the term "field" can be used instead of "beam", in an equivalent manner, especially in terms of interference.
[0042] Regarding the object, for a preferred embodiment of both the system and the method of the invention, said object is a tiny dielectric nanoparticle or equivalently biological matter such as proteins with small sizes down to 10kDa or below.
[0043] The present invention constitutes a system and a method to enhance contrast and sensitivity in scattering interference imaging.
[0044] The main purpose of the present invention is to enable the label-free detection and tracking of small [low-refractive] index single nanoparticles such as biological proteins and viruses in a simple measurement configuration.
[0045] Since scattering cross-sections scale as D6, where D is the dimension of the particle, for small particles the scattering is incredibly weak. This means that the reference beam reflection far overpowers the scattering signal. To increase the signal, one needs to increase the intensity of the incident beam, but this also increases the intensity of the reference beam. In conventional iSCAT this has meant the need to purchase fast-cameras with large full-well capacity just to be able to collect the huge number of photons produced. Such cameras are prohibitively expensive and generally with low quantum efficiency (due to detector fill-factor limitations), thwarting detection efforts. Most of the photons they detect are not due to the scattering signal but due to the reference beam. This means a lot of the shot noise and other noise on the detector is caused by the reference photons, making it harder to detect the interference signal. In the present invention, the reference beam is massively attenuated (as much as desirable), which eliminates this detector problem, as the beam can be reduced by many orders of magnitude to nearer the scattering intensity of the particles.
[0046] Further to this, the system of the first aspect of the present invention does not have any moving optical parts (such as galvo scanners or other moving optical mechanisms, which are frequently used in conventional iSCAT systems), and comprises only very simple optics, with the interference mask being the only customised optic. The uncomplicated setup with few optics, used according to the present invention, is a major benefit over existing systems, and further adds much needed stability, essential to the measurement of smaller particles.
[0047] In addition to the above described principle use, the following further configurations can be imagined, for further embodiments:
1. Plug-in module to existing commercial microscope systems: The mask and light source can easily be adapted for use in a commercial microscope setup as a plugin addition to both reflective and transmissive microscopes.
2. Fluorescence: The system of the first aspect of the invention can easily be combined with existing fluorescence microscopy to provide simultaneous fluorescence and scattering measurements.
3. Wavelength: The system of the invention can operate at multiple wavelengths.
4. Mask position: The mask can be placed at conjugate back focal planes in alternative imaging systems, if position of the mask directly below the objective is prohibitive or undesired.
5. Mask as filter: The mask can be adapted as a wavelength dependent filter to be able to combine it with attenuate a reference beam for scattering and allow fluorescence beam to pass unhindered.
6. Point-of-care implementation: For the detection of larger particles, e.g. larger proteins such as exosomes (which have been shown important for monitoring cancer activity), the setup can further be simplified to the point that it can be converted into something the size of a DVD/CD player, or even using an adapted DVD/CD or Blu-ray® player (which already contains most of the components needed in the setup) to create ultra-cheap devices which could be used at point-of-care or even in a domestic-care situation.
Industrial Applications:
[0048] The system and method of the present invention enables the detection of small changes in refractive index. It can be used in a wide range of industrial applications. The far lower cost would enable sensitive devices, previously prohibitively expensive, to be used in a wide-range of settings. Including:
Molecular Biology: The invention can be used to detect and track proteins/viruses and protein binding events down to very small proteins (∼10kDa or below). This could be used to study protein behaviour in vitro as well as to study protein movement on cell membranes.
Biomedicine: Used in combination with antibody arrays, this could be used to detect single protein binding events over large arrays for use as biomolecular detection in point-of-care settings.
Pollution: The system of the first aspect of the present invention can be implemented, for an embodiment, for portable use to detect pollution/contaminants in water supplies.
Quality control: The present invention can be used to test purity of solutions of small nanoparticles with potential to be combined into a nanoparticle sorting system.
Surface characterization: The invention can be used to characterise surface roughness on transparent surfaces or in thin film depositions such as in semiconductor fabrication.
Point-of-care: For an embodiment, the system can be incorporated into an adapted DVD/ CD/Blu-ray® player with far simpler optics to allow the detection of larger particles (the far simpler objective in a DVD/CD/Blu-ray® player would prohibit reaching ultrasensitive detection limit) which are still relevant for monitoring health conditions. Such as exosomes present in the blood stream which have recently been shown to be relevant for monitoring cancer activity of tumours in the body.
[0049] These are just a small selection of possible applications, principally based around the ability to detect very small dielectric nanoparticles both organic and inorganic.
BRIEF DESCRIPTION OF THE FIGURES
[0050] In the following some preferred embodiments of the invention will be described with reference to the enclosed figures. They are provided only for illustration purposes without however limiting the scope of the invention.
Figure 1 shows two plots taken from iSCAT paper by Piliarik et al. (2014) showing iSCAT contrast achieved for different molecular weight proteins (a) and the signal noise as a function of frame averaging (b) when the camera is run at 3000 fps, indicating the shot-noise limit. Wavelength employed: 405 nm, 10 mW at 4.5x4.5 µm field of view = approx. 50 kW/cm2.
Figure 2 shows equivalent measurements to the iSCAT measurements by Piliarik et al. (2014) performed using the system of the first aspect of the present invention. Mean contrast is plotted against protein weight (a) and signal noise as a function of equivalent camera frame rate (b). Wavelength employed: 520 nm, 33 mW at 10x10 µm field of view = approx. 35 kW/cm2.
Figure 3 schematically shows the system of the first aspect of the invention for an embodiment implementing a reflective mode arrangement, where the reference beam is reflected on an interface.
Figure 4 schematically shows the system of the first aspect of the invention for an embodiment implementing a transmissive mode arrangement, where the reference beam is transmitted through an interface.
Figure 5 shows further measurements performed using the system of the first aspect of the present invention. Detection limit is plotted against camera frame rate and compared to existing iSCAT system based on extracted published data.
DESCRIPTION OF THE PREFERRED EMBODIMENTS
[0051] Figures 3 and 4 show two alternative implementations of the system of the first aspect of the invention, particularly the above mentioned reflective mode implementation (Figure 3) and transmissive mode implementation (Figure 4), both of which relate to a common-path interferometric scattering imaging system, comprising:
illuminating means comprising a light source S configured and arranged for emitting an illumination beam L0 along an illumination optical path including two different phases of matter, one of which is constituted by the material from which the coverslip C is made (generally glass) and the other one by the medium W (in this case water) into which the objects T (in this case nanoparticles, such as proteins) are placed;
a scattered beam Ls provided by the light scattering on several objects T of a portion of the illumination beam L0; and
a reference beam Lr provided by the reflection on or transmission through an interface I of another portion of said illumination beam L0, wherein the interface I is a surface forming a common boundary surface among the coverslip C and medium W;
attenuation means comprising a partially transmissive mask M arranged in the common collection optical path for attenuating said reference beam Lr before it arrives at image sensing means D;
image sensing means D (generally including an imaging lens and a camera) configured and arranged for receiving and sensing the collected scattered Ls beam and the reference Lr beam, once attenuated by the mask M, interfering thereon as an interferometric light signal; and
processing means P connected to the image sensing means D to receive data corresponding to the interferometric light signal, and configured to process the received data to at least detect the objects T.
[0052] For both arrangements, of Figures 3 and 4, the partially transmissive mask M has a semi-transmissive first region M1 arranged in a corresponding region of the common collection path through which the reference beam Lr travels, such that the reference beam Lr is attenuated before reaching the image sensing means D, and a fully or substantially fully transmissive second region M2 arranged in a corresponding region of the common collection path through which part of the scattering beam Ls travels, such that it is traversed thereby before reaching the image sensing means D.
[0053] As shown in Figure 3, for the implementation there illustrated, the light collecting means are configured and arranged for collecting the reference beam Lr provided by the reflection on the interface I of the above mentioned another portion of the illumination beam Lo, and the system comprises an objective lens OL which forms part of both the illuminating means and the light collecting means and which is configured and arranged in both the illumination and the collection optical paths to, respectively:
focus the illumination beam Lo into the back-focal plane of the objective lens OL to produce plane-illumination out of the front aperture of the objective lens OL, such that a portion thereof will be reflected by the interface I generating the reference beam Lr and the rest will pass through the interface I up to the objects T generating the scattering beam; and
receive and at least partially collect both the reference beam Lr and the scattering beam Ls.
[0054] The objective lens OL is configured and arranged such that the reference beam Lr exits the objective lens OL as a diverging beam from the centre of the objective lens OL, when it entered as a plane wave, and impinges on the first region M1 of the partially transmissive mask M which highly attenuates it letting pass through there only a small percentage (preferably below 1 %, and more preferably around 0.1% in terms of beam intensity, or equivalently relative to field amplitude with an attenuation factor preferably below α = 0.1 or more preferably around α = 0.03) of the reference beam Lr, while the scattering beam Ls leaves the objective lens OL as a plane wave across a full back-aperture of the objective lens OL, when it entered as a spherical wave, and passes mostly through the fully or substantially fully transmissive second region M2 of the partially transmissive mask M, although a central part of the scattering beam Lr passes through the first region M1 of the mask M and is thus attenuated thereby.
[0055] As shown Figure 3, the first region M1 of the partially transmissive mask M is also placed in the illumination optical path and is configured and arranged to reflect the illumination beam L0 coming from the light source S towards the back-focal plane of the objective lens OL.
[0056] Specifically, for the reflective mode implementation of Figure 3, the system of the present invention constitutes a stand-alone microscope imaging system based on reflection scattering as described above and in more detail as follows:
1. Light of short temporal-coherence length is created by modulating the supply current of a standard laser-diode (light source S) at high frequency (>1 MHz) which is commonly implemented in consumer laser systems such as Blu-ray® players. This decreases the laser coherence length to reduce interference of objects outside of the range of interest. This is preferably as short as possible, but long enough to keep coherence between the source of the reflection as the reference beam Lr, for the illustrated embodiment, the glass-water interface between the coverslip C, and the particle T positioned on top of this. A super-bright LED or similar short coherence length light source could be used instead of modulated laser.
2. Light is focused into the back-focal plane of the objective lens OL to produce plane-illumination out of the front aperture of the objective lens OL. The interference mask M is used here as a mirror to simplify coupling of the incident beam L0 into the objective lens OL (although this can be accomplished in other ways).
3. The plane illumination beam is partially reflected from the non-index matched glass-water interface I of the objective lens OL generating the reference beam Lr. The rest of the illumination beam Lo passes through the interface I interacting with the sample T in the water W. This light principally generates Rayleigh scattering (in small particles) in-phase with the transmitted light. The scattered light can be approximated as a point-source of light emitting spherical waves propagating in all directions and this is the sample or scattering beam Ls. The phase of the scattered beam Ls is shifted relative to the incoming beam Lo due to a Gouy phase shift of pi/2.
4. The reference Lr and scattered sample beam Ls are partially collected by the same objective lens OL. The sample beam Ls leaves the objective lens OL as a plane wave across the full back-aperture of the objective lens OL since it entered as a spherical wave. The reference beam Lr (entering as a plane wave) exits the objective lens as a diverging beam from the centre of the objective lens OL.
5. The sample beam Ls impinges on the interference mask M and is mostly transmitted in the transparent regions M2 surrounding the centre, with the central part M1 of the mask M blocking only a small percentage of this beam [as in dark-field microscopy]. The reference beam Lr hits the centre region M1 of the interference mask M and is almost completely attenuated. However, importantly the centre region M1 of the mask M leaks some of the reference beam Lr through to allow for interference on the detector, i.e. on the image sensing means D (which includes at least an imaging lens and a camera).
6. The sample plane is then imaged onto a camera of the image sensing means D where the two beams Lr, Ls interfere providing the contrast to measure the particles T.
[0057] In contrast to the implementation of Figure 3, for the transmissive mode implementation of the system of the first aspect of the invention illustrated in Figure 4, the light collecting means are configured and arranged for collecting the reference beam Lr provided by the transmission through the interface I of the above mentioned other portion of the illumination beam L0, wherein:
the illuminating means comprises an illumination objective lens OLi configured and arranged to focus the illumination beam Lo into the back-focal plane of the illumination objective lens OLi to produce plane-illumination out of the front aperture of the illumination objective lens OLi, such that a portion thereof will be scattered by the objects T generating the scattering beam Ls which will be transmitted through the interface I, and another portion will be directly transmitted through the interface I generating the reference beam Lr; and
the light collecting means comprise a collection objective lens OLc configured and arranged to receive and at least partially collect both the reference beam Lr and the scattering beam Ls.
[0058] The collection objective lens OLc is configured and arranged such that the reference beam Lr exits the collection objective lens OLc as a diverging beam from the centre of the collection objective lens OLc, when it entered as a plane wave, and passes through the first region M1 of the partially transmissive mask M which highly attenuates it letting pass there through only a small percentage (preferably below 1 %, and more preferably around 0.1% in terms of beam intensity, or equivalently relative to field amplitude with an attenuation factor preferably below α = 0.1 or more preferably around α = 0.03) of the reference beam Lr, while the scattering beam Ls leaves the collection objective lens OLc as a plane wave across a full back-aperture of the collection objective lens OLc, when it entered as a spherical wave, and passes mostly through the fully or substantially fully transmissive second region M2 of the partially transmissive mask M, although a central part of the scattering beam Lr passes through the first region M1 of the mask M and is thus attenuated thereby.
[0059] Specifically, for the transmissive mode implementation of Figure 4, the system of the present invention constitutes a stand-alone microscope imaging system based on transmission scattering as described above and in more detail as follows.
[0060] In transmissive-mode, the microscope operates mainly the same as in reflective-mode. More optics (principally a second objective) are required as a new excitation path from above the sample is needed.
[0061] The principle remains the same as that in reflection with a reference beam Lr and scattering signal beam Ls generated by a single excitation light source S then interfere on a detector D after the reference beam Lr is partially attenuated by a partially transmissive mask M or equivalent.
[0062] The main difference here is that the reference beam Lr will be much stronger in intensity, as here it is almost 100% the intensity I0 of the incident beam Lo, as most of the light is transmitted rather than reflected by the interface I. This differs in the reflective-mode case, as in that case the reference beam Lr is generated by the reflection off the glass/water interface I, which reduces its intensity to around 0.5% of the initial beam Lo.
[0063] In practice this means that in transmissive mode, the mask M must attenuate the reference beam Lr by at least one order of magnitude more compared to the reflective mode case. This potentially complicates the production of the mask M. Along with simpler optics, this highlights the distinctive benefit of the reflective mode case where the reference beam Lr is pre-attenuated by the glass/water interface I. However, given a suitable mask, both are equivalent.
Mask construction:
[0064] Regarding the above described mask M, it was built for its inclusion in the system of the present invention, for an embodiment (for the arrangements of Figures 3 and 4), as follows:
1. The mask M consists of a semi-transmissive section and a transmissive section.
2. The semi-transmissive section M1 was created by depositing metal onto a sacrificial premask defining the semi-transmissive region on an optical flat.
3. A vinyl sticker was used as a pre-mask to define the area.
4. Metal was evaporated at the desired thickness to attenuate signal passing through, ensuring metal was evenly deposited.
[0065] The mask M itself can be constructed in many different forms and materials depending on availability and exact implementation needed. A well-formed mask with precise thickness is key to obtaining reliable and symmetric interference patterns on the detector.
[0066] Specifically, as stated in a previous section, for an embodiment (not shown), the collection of both the reference and the scattering beams is performed on reflecting from the mask, the latter having a semi-reflective section (almost transparent) for the reference beam (thus attenuated by reflection) and a reflective section for the scattering beam.
[0067] Also, for the manufacturing of the mask, instead of metallic coatings, dielectric anti-reflective/reflective Bragg type coatings can be used, for other embodiments.
[0068] The technique used in the system and method of the present invention significantly improves on the published conventional iSCAT technique (described in the Background section above) while allowing better contrast and sensitivity. Here the benefit of the technique of the present invention over iSCAT, the best implementation as yet of interferometric light scattering microscopy, is elaborated.
[0069] In general, in interference scattering microscopy, the signal imaged onto the detector has intensity:
[0070] Where, as stated in a previous section, r is a co-efficient describing the amplitude of the reference beam, s is a co-efficient relating to the amplitude of the scattering signal, and θ is the phase difference between the two signals. For detecting small particles such as proteins the difference between r2 and s2 is many orders of magnitude (around 107 for a 100 kDa protein) making it practically impossible to measure the scattering signal upon the background of the reference beam. Crucially the interference term, scales both with r and s, meaning there is much less difference between this and the r2 term, only around 104 for the same 100kDa protein. This then becomes possible to measure with the latest detectors and very stable light source coupled with low noise levels.
[0071] This key advantage of the system and method of the present invention, is the in-line suppression of the reference signal relative to the scattering signal in an in-line interference microscopy setup similar to iSCAT. This allows the optimisation of the contrast between the reference beam intensity and the interference cross-term. This enables the dramatic reduction of the unwanted reference beam intensity relative to the interference intensity and thus increase the sensitivity of the setup and reduce dependency on noise and stability of the excitation light source and overall setup. Since far fewer photons overall are now being detected, the camera used can be replaced with far cheaper versions as the huge dynamic range is no longer needed. It also means that a very cheap laser or LED light source with short coherence length can be used.
Comparison to conventional iSCAT:
[0072] With reference to Figures 1 and 2, the technique of the system and method of the present invention is directly compared to the best achieved in the literature taken from Piliarik et al. (2014). They use iSCAT to detect the binding of single proteins of various sizes from 65.5 kDa (BSA) to 340 kDa (fibrinogen). Using their noise statistics it is calculated how long they need to integrate for to detect their smallest protein (BSA), i.e. to detect the signal (iSCAT contrast) above the noise. For the BSA protein the molecular weight is 38 kDA and from Figure 1 (a), this corresponds to an iSCAT contrast of 3x10-4. From the noise statistics, Figure 1 (b), they need to average over 600-700 frames. Since they run their camera at 3000 fps, this corresponds to 0.2 s integration or equivalently running at 5 fps, which is the minimum speed they can run and detect/track BSA protein.
[0073] The present inventors repeated similar experiments using the system of the present invention (Figure 2), for the arrangement of Figure 3. It can be seen that to detect BSA (see Figure 2(a)) given the signal noise measured in the present experiment it can run at a higher rate of 60fps (see Figure 2(b)). This is more than an order of magnitude (∼x12) improvement in sensitivity.
[0074] Further experiments were performed by the present inventors with the system of the first aspect of the invention, particularly for non-specific binding of a variety of single proteins to a coverglass in comparison to a control with buffer only. Figure 5 shows the results of said further experiments, where detection limit is plotted as a function of frame averaging (dots) in comparison to shot-noise-limited behaviour (dashed line). The acquisition rate for the experiments was 400 FPS. The dotted line and triangles show a comparison to the detection limit extracted from Piliarik et al. (2014).
Key Advantages of the system and method of the present invention:
a) Increased signal level
Scattering intensity scales inversely with the fourth power of illuminating wavelength, and the interference cross term [2rs cos θ] scales inversely with the square of illuminating wavelength. Since in Piliarik et al. they used a shorter wavelength and more powerful laser, actual gains of the system and method of the present invention are higher than an order of magnitude. If parameters identical to those reported previously (wavelength and intensity) were to be employed in the present invention, signal sensitivity would increase by another factor of 2.4. Thus with a total improvement in sensitivity of around 30.
b) Reduced sensitivity to reference beam instability
The attenuation of the reference beam according to the present invention reduces the effect of instability in phase and intensity in this signal introduced throughout the beam path or from the laser and spatially across the field of view in the system/microscope. This allows to move to larger field of view on the system/microscope thus detecting more particle binding sites at once. No deterioration was noticed in signal moving from the 10x10 µm field of view shown in Figure 2, to 40x40 µm (16x bigger area). This is a large advantage over conventional iSCAT which struggles with even detection of gold nanoparticles in a wide-field setup (i.e. without using Galvometric scanning or other scanning techniques) [see Opt. Express 14, 405 (2006)], due to its extreme sensitivity to phase shifts from interfering back reflections.
c) Cost
The increased signal, lower photon count on the detector and increased stability of the signal lead to a setup which for the same level of detection costs far less to implement and requires a simpler geometry. The detectors, light source and other optical elements in a conventional iSCAT setup, typically put the cost at > $150,000, while in the proposed setup for the system of the present invention, the purchased elements can easily be found for < $10,000. With most of this cost due to the objective lens. With further modifications it is feasible to imagine a system costing even less and at the cost of sensitivity objective could be massively simplified for systems in the sub-$2000 range.
[0076] These advantages clearly illustrate the unique nature of the system and method of the present invention and the large impact it could have in industry.
[0077] A person skilled in the art could introduce changes and modifications in the embodiments described without departing from the scope of the invention as it is defined in the attached claims.
1. A common-path interferometric scattering imaging system, comprising:
- illuminating means comprising a light source (S) configured and arranged for emitting an illumination beam (L0) along an illumination optical path including at least two different phases of matter;
- light collecting means configured and arranged for simultaneously at least partially collecting through a common collection optical path:
- a scattered beam (Ls) provided by the light scattering by an object (T) of a portion of said illumination beam (L0), wherein said object (T) is placed in at least one of said two different phases of matter; and
- a reference beam (Lr) provided by the reflection on or transmission through an interface (I) of another portion of said illumination beam (L0), wherein said interface (I) is a surface forming a common boundary among said two different phases of matter;
- image sensing means (D) configured and arranged for receiving and sensing the collected scattered (Ls) and reference (Lr) beams interfering thereon as an interferometric light signal; and
- processing means (P) connected to said image sensing means (D) to receive data corresponding to said interferometric light signal, and configured to process said received data to at least detect said object (T);
characterised in that it comprises attenuation means arranged in said common collection optical path for attenuating said reference beam (Lr) before it arrives at said image sensing means (D).
2. A system according to claim 1, wherein said attenuation means comprises:
- a partially transmissive mask (M) having a semi-transmissive region arranged in a corresponding region of the common collection path through which the reference beam (Lr) travels, such that the reference beam (Lr) is attenuated on transmission before reaching the image sensing means (D); or
- a partially reflective mask (M) having a semi-reflective region arranged in a corresponding region of the common collection path through which the reference beam (Lr) travels, such that the reference beam (Lr) is attenuated on reflection before reaching the image sensing means (D).
3. A system according to claim 2, wherein said semi-transmissive or semi-reflective region is a first region (M1) of said partially transmissive or partially reflective mask (M), the mask (M) comprising a second region (M2) arranged in a corresponding region of the common collection path through which part of the scattering beam (Ls) travels, such that said part of the scattering beam (Ls) traverses said second region (M2) or is reflected thereon before reaching the image sensing means (D), by transmission or by reflection, wherein said first (M1) and said second (M2) regions have different transmissive or reflective properties and said partially transmissive or partially reflective mask (M) maintains the coherence relationship between the reference (Lr) and scattered (Ls) beams.
4. A system according to claim 3, wherein said second region (M2) is a fully or substantially fully transmissive or reflective region.
5. A system according to claim 3 or 4, wherein said first region (M1) has a circular or cylindrical shape and said second region (M2) has an annular or tubular shape with an inner diameter larger than the diameter of said first region (M1) and being arranged concentrically with respect thereto.
6. A system according to any of the previous claims, wherein said illumination optical path and said common collection path are configured and arranged such that the reference (Lr) and scattered (Ls) beams are generated at such closer positions that ensure a phase-locked relationship between the reference (Lr) and scattered (Ls) beams, the system being absent of any phase varying mechanism for said reference (Lr) and scattered (Ls) beams.
7. A system according to any of the previous claims, comprising a coverslip (C) for said object, wherein said interface (I) is the common boundary surface among said coverslip (C) and a medium (W) into which said object (T) is placed, the material of which said coverslip (C) is made being non-index matched with said medium (W).
8. A system according to any of the previous claims, wherein said light collecting means are configured and arranged for collecting said reference beam (Lr) provided by the reflection on said interface (I) of said another portion of the illumination beam (L0), wherein the system comprises an objective lens (OL) which forms part of both the illuminating means and the light collecting means and which is configured and arranged in both the illumination and the collection optical paths to, respectively:
- focus the illumination beam (L0) into the back-focal plane of said objective lens (OL) to produce illumination out of the front aperture of the objective lens (OL), such that a portion thereof will be reflected by the interface (I) generating the reference beam (Lr) and the rest will pass through the interface (I) up to the object (T) generating the scattering beam; and
- receive and at least partially collect both the reference beam (Lr) and the scattering beam (Ls).
9. A system according to claim 8 when depending on claim 3, wherein said objective lens (OL) is configured and arranged such that the reference beam (Lr) exits the objective lens (OL) as a diverging beam from the centre of the objective lens (OL), and passes through or is reflected on the first region (M1) of the partially transmissive or partially reflective mask (M), and the scattering beam (Ls) leaves the objective lens (OL) as a plane wave across a full back-aperture of the objective lens (OL), when it entered as a spherical wave, and passes through or is reflected on both the first (M1) and the second (M2) regions of the partially transmissive or partially reflective mask (M).
10. A system according to claim 9, wherein said first region (M1) of the partially transmissive or partially reflective mask (M) is also placed in the illumination optical path and is configured and arranged to reflect the illumination beam (Lo) coming from the light source (S) towards the back-focal plane of the objective lens (OL).
11. A system according to any of claims 1 to 7, wherein said light collecting means are configured and arranged for collecting said reference beam (Lr) provided by the transmission through said interface (I) of said another portion of the illumination beam (Lo), wherein:
- the illuminating means comprises an illumination objective lens (OLi) configured and arranged to focus the illumination beam (L0) into the back-focal plane of said illumination objective lens (OLi) to produce plane-illumination out of the front aperture of the illumination objective lens (OLi), such that a portion thereof will be scattered by the object (T) generating the scattering beam (Ls) which will be transmitted through the interface (I), and another portion will be directly transmitted through the interface (I) generating the reference beam (Lr); and
- the light collecting means comprise a collection objective lens (OLc) configured and arranged to receive and at least partially collect both the reference beam (Lr) and the scattering beam (Ls).
12. A system according to claim 11 when depending on claim 3, wherein said collection objective lens (OLc) is configured and arranged such that the reference beam (Lr) exits the collection objective lens (OLc) as a diverging beam from the centre of the collection objective lens (OLc), when it entered as a plane wave, and passes through or is reflected on the first region (M1) of the partially transmissive or partially reflective mask (M), and the scattering beam (Ls) leaves the collection objective lens (OLc) as a plane wave across a full back-aperture of the collection objective lens (OLc), when it entered as a spherical wave, and passes through or is reflected on both the first (M1) and the second (M2) regions of the partially transmissive or partially reflective mask (M).
13. A system according to any of the previous claims, wherein said processing means (P) implements an algorithm to process the received data according to the following equation:
where r is the normalised reference beam amplitude, s is the normalised scattering beam amplitude, θ is the phase difference between the reference (Lr) and scattering (Ls) beams, α is the attenuation amplitude defined as the reciprocal transmission amplitude, Itotal is the total intensity of the light on the image sensing means (D) caused by the two interfering reference (Lr) and scattering (Ls) beams, and I0 is an initial light intensity on the image sensing means (D), wherein said attenuation means have a degree of attenuation for said reference beam (Lr) calculated with the purpose of maximizing the term 2rs cos θ with respect to the term r2 of the above equation to enhance detection sensitivity.
14. A method of using common-path interferometric scattering imaging to detect an object, comprising:
- emitting an illumination beam (L0) along an illumination optical path including at least two different phases of matter;
- simultaneously at least partially collecting through a common collection optical path:
- a scattered beam (Ls) provided by the light scattering on an object (T) of a portion of said illumination beam (L0), wherein said object (T) is placed in at least one of said two different phases of matter; and
- receiving and sensing, on image sensing means (D), the collected scattered (Ls) and reference (Lr) beams interfering thereon as an interferometric light signal; and
- receiving and processing data corresponding to said interferometric light signal to at least detect said object (T);
characterised in that it comprises attenuating said reference beam in said common collection optical path before it arrives at said image sensing means (D).
15. A method according to claim 14, wherein said object (T) is a dielectric nanoparticle with a size of substantially 10kDa or below.
Non-patent literature cited in the description
LINDFORS et al.PRL, 2004, vol. 93, 3- [0005]
PILIARIK et al.Nature Communications, 2014, vol. 5, 4495- [0005]
Opt. Express, 2006, vol. 14, 405- [0075]


Countdown the Board Game
Countdown is one of the longest-running and most iconic quiz shows on British television.With its distinctive music and compulsive game-play, the show has recently been given a new lease of life - and audience - through the 8 Out of 10 Cats Does Countdown adaptation on Channel 4.With this renewed popularity, Rocket Games has released a brand new version of the Countdown board game featuring music from the show and all your favourite rounds.Now you can play along as the stars do and hope to achieve a nine letter word or the elusive target number before the 30 seconds expire.There's also the Countdown Conundrum, where you need to be quick off the mark to spot the jumbled up word before your fellow contestants.And for a real feeling of being there, the familiar music will count you down so you don't have to take your eyes off the page as you try to win the game that has become a true household name since it launched Channel 4 back in 1982. Show more Show less


The Reality of Being a Gamer Of Color in eSports
FEATURED, GAMING, TECH
Editor's Note: This is a guest post from Jeremiah Timmons. Jeremiah is an avid gamer and somewhat less avid in his interactions with other human beings. He writes for various publications, screenplays, as well as his personal blog Fukette.com
Terrence "TerrenceM" Miller At DreamHack (Mulkerin,2016)
If you were to travel back to 2005 and approach a gaggle (for the sake of this piece, a group of players will exclusively be referred to as a 'gaggle') of teenaged gamers with the prospect of gaming AND getting paid to do so, I'm confident in saying that at least one, if not all, of them, will call you a liar. Well, that and a whole host of other obscenities one can't help but learn just by spending a few hours play playing against other depraved souls online. The mere concept of participating in a competition against other gamers from around the world while earning more money than most people make in an entire year is incredibly exciting. And why shouldn't it be? Not only will you get the chance to dive into the abyss when it comes to the games you know and love, but you'll also get to meet like-minded individuals that are hopefully as obsessed as you are.
At least, that would be an IDEAL scenario.
African Gaming League In Nigeria (Takahashi, 2016)
When it comes to the reality of participating in professional leagues, gamers of color are subjected to experiences that are very different than what would be considered ideal. Where most people would expect to find others with compatible interests and somewhat similar sensibilities, gamers of color are reporting discriminatory practices that seem to go out of its way to target players whose demographics are scarce, if not completely non-existent, to begin with.
An example would be the constant barrage of racial slurs and insults hurled at black players through platforms like Twitch whenever they compete as evidenced by Terrence "TerrenceM" Miller's ordeal as he competed at the DreamHack Austin tournament last May. Viewers of his matches, which included his parents, had started to notice that the chat window had become blanketed with racial slurs all directed at Terrence. Unfortunately, with the exception of his family, Terrence had steeled himself in preparation for dealing with racist trolls because he's long since learned to expect these things.
"Obviously, the fact that it was happening bothers me, but hearing that people are saying racist things about me on the Internet is nothing new, and it's not surprising that Twitch chat was doing that." Miller says.
As even the most casual of observers will note, gaming, with eSports being a highly visible example, can be a very hostile and combative environment where insults and even threats are considered commonplace more than isolated incidents. Often, POC, women, and generally gamers who are known or even rumored to be homosexual or anyone whose demographics are relegated to the margins of what is considered 'normal' will get inundated with comment threads ranging from ethnic slurs, homophobic threats, and even statements threatening female competitors with rape. Couple this with the fact that most tournaments simply have no protocol in place to mitigate this disgusting behavior if only because it affects such a small percentage of its players and the cost-benefit ratio just does not support such efforts at this time. How tragic.
The Real Stars Multi Gaming Organization
With the current climate in professional gaming being the unabashed racism and prideful ignorance exhibited by mainly cis, white males, there's hardly any confusion as to why the African-American presence in the professional league circuit is almost non-existent. According to the findings of The Kaiser Family Foundation, African American children between the ages of 8 and 18 play games 30 minutes more per day than white children, while Hispanics play an average of 10 minutes more than white children. It also states that African-American and Hispanic children are more likely to purchase games than white children. (Source) That would indicate that children of color are just as enamored with the concept of gaming as much as their white counterparts, but are frequently subjected to barriers white players could not even imagine much less have to contend with.
When asked about what can be done to combat such behavior from his fellow competitors, as if the task of curbing vicious hate-speech from other humans with developed brains and at least a cursory knowledge of American history is incumbent upon him, Terrence Miller responded with "Just ignoring it like some people are saying doesn't make the problem go away. It'll just keep happening. You have to keep bringing it up and try to find a resolution."
It goes without saying that this is an incredibly eloquent response considering the context.
eSports Arena
However, real change must come from the organizers and sponsors who responsible for hosting these tournaments. One really can't rely on substantive change being employed as a result of a moral imperative, but one can always count on organizations taking heed from those who are its customers, the buying public. The amount of money that could be amassed by making the tournament scene accessible to players from all walks of life is staggering in estimation, so it begs the question: Why would you allow your negligence regarding such a serious issue deprive you of so much in terms of both financial and PR capital? Encouragingly, the DreamHack staff has publicly stated that they are "taking active steps to improve for DH Summer." While the measures have not been explicitly outlined, one can only assume that whatever action taken has to be better than the laissez-faire attitude taken by most organizations up til now.
Tags : African Gaming League, eSports, Gaming, Terrence M, Video Games, videogames
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