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PROVIDENCE - The game changed on an alley-oop.
Rhode Island freshman Jordan Hare rose above the Providence College defense to emphatically finish a perfectly executed play with 15 minutes, 22 seconds left in the second half to cut the Friars' lead to five.
As it turned out, the alley-oop was the best thing that happen to PC because Friar coach Ed Cooley opted to eschew his zone defense against the athletic Rams and play man-to-man. Over the next 8 minutes, 21 seconds the Rams didn't make a basket, went 0-for-8 from the field and committed six turnovers in a 72-57 defeat.
"They played really hard. They're well coached and we were just really fortunate to come away with a win," Cooley said. "... For all the coaches and whoever's listening, our players have done an unbelievable job."
Providence College (7-2) was led by LeDontae Henton, who produced 21 points and a game-high 17 points. Kadeem Batts scored a game-high 23 points, while Bryce Cotton played through a knee injury to score 11 points and dish out seven assists as the Friars picked up their fifth straight win overall.
Rhode Island (2-7) received an efficient effort from Hare, who scored 14 points on just seven shots. Freshman wing Xavier Munford, URI's leading scorer had one of the worst games of his young career, going 3-for-13 - including 0-for-8 from the 3-point line - on his way to a 10-point, three-turnover effort.
'We turned the ball over 20 times against someone that plays a zone defense in the half court for the majority of the game, that's disappointing," URI coach Dan Hurley said. "Our inability to rebound the ball has been an issue for us through out the year. We have a real thin margin for error to score the ball."
Both teams came into Thursday's rivalry game knowing better times lay ahead and that's probably the only solace Hurley cane take away from the game because his team did little right for the second straight trip to the Dunkin' Donuts Center. The Rams were a horrid 3-for-18 from the 3-point line and committed 20 turnovers against just 11 assists one game after producing a positive assist-to-turnover ratio for the first time this season.
Just as they have done all season, the Friar starters did everything. The combination of Batts, Henton, Josh Fortune and Cotton combined to score all of PC's points.
The first half was a cagey, tight affair with neither team able to take control of the game. PC stuck to its zone defense, while the Rams played a tight man-to-man defense. The Friars, behind 10 points from Batts, took a one-point lead into halftime after Rhody grabbed a late first-half lead on a Mike Aaman basket in the paint.
The Rams produced a 5-0 run - punctuated by a Nikola Malesevic hoop in transition - to take a two-point lead, the visitors largest of the night. The lead would also turn out to be the Rams' last because PC went on an 11-2 run to take complete control of the game.
Hare closed the gap to just five with his dunk after a perfect pass from Munford on the left wing.
Cooley then changed to a man-to-man defense against the more atheistic Ram lineup and when Hurley elected to insert Aaman, T.J. Buchanan and Alwayne Bigby into the lineup Cooley went back to the 2-3 zone defense and dared the Rams to make outside shots.
"When we saw them go man we were excited because we liked our match ups," Hurley said. "We turned the ball over 20 times against a team that doesn't pressure and that just can't happen."
URI's turnovers during its barren spell led to Providence College baskets. Following a Mike Powell turnover Fortune scored in transition and the Friars scored a few more buckets following turnovers to open up a 52-39 lead before Malesevic hit a jumper to stop the bleeding with 7:01 left in the game.
"He's a good coach and me made an adjustment and we had to counter the adjustment," Cooley said. "Normally we play zone and that's what they prepared for. (Hare) is a special player for them. I love that kid. Hare and Malesevic were the guys we were trying to match up with."
The Friars put the game away at the foul line as the home side made 18 of its 19 free throws in the second half. Fortune made four free throws in the final 1:18, while Henton, Cotton and Batts also made their free throws to ice the Friars' third straight win over the Rams and fifth straight at the Dunk.
Both teams will have plenty of time to regroup from Thursday's contest. The Friars don't play again until Dec. 18 when Colgate comes to town, while the Rams are off until Dec. 15 when Larry Brown and SMU come to the Ryan Center.
"I think [the break] is a great thing because we can really self study and really do a good job," Hurley said.


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2015 Community Fundraising
Thinking about fundraising for the LIVESTRONG Foundation? You've come to the right spot! Whether you want to hold a small local event, run a 5K or marathon, ride your bike, throw a party, fundraise with your company, or share a new idea with our team, we can help!
Fundraise for LiveStrong

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- Northern New York Newspapers
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HOGANSBURG Officials with the St. Regis Mohawk Tribe and Akwesasne Mohawk Casino have a ribbon-cutting media event set for Thursday to unveil the casinos new $74 million expansion.
The first addition many will notice as they pass on Route 37 is the new 150-room hotel that is attached to the casino. Officials had previously said that it will include a spa, pool and fitness area. Next door, Tarbell Management Group worked on a 37-room expansion to the existing Comfort Inn & Suites Akwesasne, providing a total of 101 rooms there. In total, there will be about 250 hotel rooms in the vicinity of the casino in 2013 as opposed to the 64 rooms that had been available at the Comfort Inn.
The Mohawk Bingo Palace, which the tribe has operated for about 27 years, relocated to a 30,000-square-foot expanded area in the casino in March.
In November, the casino launched its new Cascades gaming and entertainment area, a 40,000-square foot expansion to the existing facility. It includes new slot machines and table games as well as a high-end bar and a high-stakes gambling room with $50 per play slot machines.
Casino General Manager Patrick Bassney previously stated that the finished casino and hotel will employ about 900 people.
In March of last year, the tribe signed a $110 million deal with Key Bank to pay for the project as well as to refinance existing casino debt.
Tribal Chief Ronald LaFrance previously said the tribe had about 90 assistance programs available when the casino opened in the late 1990s, but now, with the extra gaming monies, that has expanded to more than 200.
He said he would like to see some of the new revenues go toward heating fuel assistance, scholarships, drug prevention education for the public and drug preventative training for police. He added that the Hogansburg-Akwesasne Volunteer Fire Department will be getting a $1 million ladder truck, courtesy of casino revenues.
Mr. LaFrance and Mr. Bassney each declined Wednesday to comment on the project prior to the ribbon cutting.
Thousands of readers like you have made Northern New York Newspapers the number one ranked website* for local news and sports coverage in the north country.
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Coverall Pressed Powder
This beauty bargain gets the job done in terms of setting makeup and adding light coverage.
Color Icon Bronzer SPF 15
If you?re looking for something that does the job at a good price, Color Icon Bronzer might be worth checking out.
Add to Faves?
Wet 'n' Wild At-A-Glance
Strengths: Inexpensive, good tinted moisturizer with sunscreen; some attractive bronzing powders; mostly good eyeshadow and lipstick options.
Weaknesses: Unimpressive concealers; large assortment of average to poor eyelining products; the mascaras do little to impress; some lip products suffer from the inclusion of irritants; the makeup brushes.
Wet 'n' Wild is one of the few cosmetics companies around that prides itself on being extraordinarily cheap, although the prices have increased across the board since we last revisited this brand. But don't let its hokey name and pre-teen marketing tactics deter you, because there are some great options available, and the packaging has improved in almost every category. The best-performing products will seem like steals, while the less impressive to horrible products will quickly have you thinking, "you get what you pay for." It is not very often that a line offers such a procession of clear-cut winners and dismal losers, but once you know what to focus on, you won't get soaked by Wet 'n' Wild. This line is available in most major drugstores across the country, and, like most drugstore lines, does not offer testers. Interestingly, the selection of Wet 'n' Wild in Canadian drugstores tends to be better than what U.S. shoppers will find.
For more information about Wet 'n' Wild, visit www.wnwbeauty.com.
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East Midlands
Find other Netmums members in Derbyshire, Leicestershire, Lincolnshire, Nottinghamshire or Northamptonshire. These boards are an extension of the Netmums Meet a Mum boards which can be found on your local netmums site. You can post messages looking for new friends, arrange to meet up with other members or join in with the new Official Netmums Meetups here.
Take a look at your local Netmums site for all things local:
• make friends through our Meet a Mum board - over 1,000 new friendships are made there every month.
• find a local Official Netmums Meetups group - run by Netmums Volunteers all over the UK.
• use the noticeboard to post any queries or notices.
• visit the Places To Go section for everything from toddler groups and preschool classes to what's on at the weekend and classes for older children and adults.
• browse through all our local information.


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The Return of the Dead, by Claude Lecouteux
By Psyche | 1 comment
The Return of the Dead: Ghosts, Ancestors, and the Transparent Veil of the Pagan Mind, by Claude Lecouteux, translated by Jon E Graham, afterward by Regis Boyer Inner Traditions, 9781594773181, 273 pp. (incl. notes and index), 1996, 2009 A former professor of medieval literature and civilization at the Sorbonne, this is Claude Lecouteux's second book to be translated into English, the first being Witches, Werewolves, and Fairies: Shapeshifters and Astral Doubles in the Middle Ages, published in 2003 also by Inner Traditions.The Return of the Dead is a scholarly survey of pre-Christian beliefs, focusing in particular on legends of revenants in Germanic and Scandinavian folklore. Read More


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Ever wonder where the Rockefeller Plaza Christmas tree comes from? This year it was felled from the home of Joseph Balku in Flanders, N.J. And driven over to the Big Apple:
A day ago Joseph Balku's Norway Spruce stood 80-feet tall outside his home in Flanders, N.J. Today it is 50 miles away, the new holiday centerpiece of New York City's Rockefeller Plaza (also home to NBC News).
Over the next two weeks, the tree will be decorated with more than 30,000 multi-colored lights on 5-miles of wire. The 80-year old tree has added meaning, it has been dubbed a "Sandy survivor," coming from an area affected by the recent storm that wreaked havoc across the Northeast. According to AP:
Balku says he watched the tree, which weighs 10 tons and is 50 feet in diameter, as it swayed in the backyard of his Mount Olive property. Balku says it had been bundled for protection. He lost two other trees and just got electricity back over the weekend.
Good idea bundling the tree. Let's hope he did the same with his mother-in-law.


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The Director's Playhouse (4 month Character Development Intensives)
The Director's Playhouse
Weekly Event - Every Wednesday & Friday: 7:00 AM to 12:00 AM (PDT)
Los Angeles, CA
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The Director's Playhouse | 11543 West Olympic Blvd | Second Floor | Los Angeles | CA | 90064
In order to purchase these tickets in installments, you'll need an Eventbrite account. Log in or sign up for a free account to continue.


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Craft and work station, a tiny indoor forest and faux taxidermy. We're all over the place this week for our Friday Favorites, but we're certainly amusing and you've been telling us that you love our choices. We aim to please!
Alison's Pick
Redwood Forest Growing Kit, Overstock $48.49
Serenity now. I'm missing a green thumb so while I find joy in the thought of growing my own indoor forest and using the adorable scissors to trim it's lush greenery that couldn't house a hobbit; in truth I'd probably kill it. But as a gift this item is top notch and it comes in a steel box with blue ribbon so no wrapping required.
Christina Moffitino's Pick
Metallic Bark Pillar Candles, $23 Pottery Barn
Creating a winter world in your home is as easy as dimming the lights and sparking a candle. I'm really getting ready for the winter season by investing in chic holiday decor. They're perfect for holidays and still stylish enough to keep by bedside all year long. These candles create the atmosphere to pour yourself a cup of cocoa and relax to some good ol' Christmas music.
Lauren's Pick
Murphey Wall Mount Craft Desk, $140 Overstock
If you're like me and you have a passion for the crafts and/or you're trying to save a few pennies for the holidays by doing handmade gifts, then your room is probably a MESS! This murphy craft desk is a huge space saver and keeps all my artsy knickknacks all in one place. I use it for sewing, scrapbooking, stationary, jewelry, or sometimes just writing. It's the perfect gift for DIY-ers and crafty kids in the dorms.
Christina Wright's Pick
White and Turquoise Deer Head, $110 Etsy
I need to find a way to convince my husband that we must have this faux taxidermy deer on our wall!! I am building a collage of frames in the living room, and this would be a perfect accent piece to add a bit of whimsy to the room. It's my favorite color too! Modern taxidermy, without actually killing animals...I like it!


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Bagley caption contest: In your own words
Published May 4, 2013 7:45 pm
This is an archived article that was published on sltrib.com in 2013, and information in the article may be outdated. It is provided only for personal research purposes and may not be reprinted.
Submit a caption to go with this cartoon drawn by Tribune political cartoonist Pat Bagley.
Contest rules: To enter, submit your caption by e-mail to email@example.com, by FAX to 801-257-8525, or by postcard addressed to: Tribune Caption Contest, Suite 700, 90 South 400 West, Salt Lake City, UT, 84101.
All submissions must include the caption writer's name, home address and daytime telephone number. Entries must be received by midnight of the Tuesday following the Sunday on which a new captionless cartoon is published. Any received after the deadline will not be considered.
The writer of the winning caption will be mailed the original cartoon, signed by Bagley, with the caption included.
Reader comments on sltrib.com are the opinions of the writer, not The Salt Lake Tribune. We will delete comments containing obscenities, personal attacks and inappropriate or offensive remarks. Flagrant or repeat violators will be banned. If you see an objectionable comment, please alert us by clicking the arrow on the upper right side of the comment and selecting "Flag comment as inappropriate". If you've recently registered with Disqus or aren't seeing your comments immediately, you may need to verify your email address. To do so, visit disqus.com/account.comments powered by Disqus
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I used to have a Razer Blackwidow Ultimate, and now own a Steelseries Apex. The latter isn't mechanical, and honestly, I prefer it.
I used to have a Razer Blackwidow Ultimate, and now own a Steelseries Apex. The latter isn't mechanical, and honestly, I prefer it.
I have this in cherry blue keys. Best keyboard I think I've ever used.
EVE Online
Find me on twitter! @nikkonasty
I have a Black Widow Ultimate.
It's a nice keyboard.
Terrible value for money, and if you're looking for value there are significantly better keyboards out there.
However, if you like the look, and enjoy using it, it's a very nice keyboard.
It just seems to me that it's different for the sake of it. I want my keyboard to have proper ergonomics. Actually that includes springs. Of course a 20$ keyboard isn't going to deliver that, but a 400$ keyboard built around tech from the 90s isn't either (just looking at those keys makes my fingers hurt, lol).
Last edited by Ronin; 02-04-2014 at 03:05 PM.
Stay frosty.
The corsair raptor k30/series is great and affordable. Back lit and no ghosting as well
As opposed to...
There are other mechanical cherry types, but these are the most common. It should be clear that the last 4 have a massive advantage over the first.
got one of these babies ... because when it comes to keyboard and mouse ... logitech is the best for me <3 but for gaming pad i use ... Razer Nostromo perfect for FPS games because of really nice reaction time on jumping and etc ...
Well, my keyboard does not require me to press the key through to the bottom to register the key.
From an engineering standpoint I would be very curious as to what the advantage of using a metal spring over a plastic one is. I am not sarcastic in that. If you really wanted to you could make a plastic keyboard just as noisy as a mechanical on, I bet. It's just there is no reason to.
Stay frosty.
Most have two stages. A light tap will trigger it, however, you can bottom it out too. There's basically some buffer zone to it.
I'd really advise you to try one before being so quick to judge. I didn't think there was much different myself.
Then I got one, and well, honestly, I can't see myself buying another rubber dome.
It just feels significantly different, more comfortable.


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One of my favorite mystery authors is Henning Mankell who penned a great many international best-sellers, and is considered to be one of the leading proponents of the Nordic noir novel. Twenty-six of his books featured the policeman character Kurt Wallander and some took place in both Sweden and Mozambique. My favorites include The Fifth Woman, The White Lioness, Sidetracked and The Troubled Man.
His Wallander character was played on BBC television by Kenneth Branagh. Quite a few of his novels also featured Wallander's daughter Linda, and explore the issue of racism.
If you haven't read any of his works, do yourself a favor and check a title or two out from KPL's Central location. His books are intriguing and you might find yourself totally immersed in his characters, locale, and details. They are that good!
Although he was born in Sweden in 1948, Mankell also spent considerable time in Mozambique. Sad for all of us who love his work, he died of cancer on Oct 5, 2015 at the age of 67.
I admit to not knowing much about the Falkland Islands, the setting for the novel Little Black Lies. But the Falklands are a strong presence in this suspenseful story by S.J. Bolton, and I certainly feel as though I have a stronger sense now of the islands. In the story, three children have gone missing in this wild and beautiful place, over a period of several years. Most of the islanders feel that accidents claimed the children- perhaps a fall, or swept away by a strong tide. As events unfold and the main characters and motives are revealed, it becomes apparent that certainly not all of the disappearances can be explained away by accidents. Strong characters, a fast paced story, and a fascinating setting make Little Black Lies a winner. It was recommended to me by a co-worker, who said he feels it's one of the best books he's read all year, and I agree.
This is sort of a fun read for those who may be looking for
a bit of a darker read but aren't really ready for something scary. The head mistress of a ladies' finishing
school and her brother are poisoned and rather than report the crime to the
local police the seven students decide to hide it in an effort to avoid being
sent home and separated from each other.
Disgraceful Mary Jane, Sly Kitty, and Stout Alice (each girl has a
moniker) haphazardly cobble together a cover up while Pocked Louise sets her
sights on finding the killer. The
Scandalous Sisterhood of Prickwillow Place is an interesting read for anyone
who enjoys murder mysteries with female protagonists.
I was delighted to hear that author Alan Bradley had a new book. As Chimney Sweepers Come to Dust: a Flavia de Luce Novel features twelve year old Flavia's adventures in Canada in the 1950's, where she has been sent to boarding school. It is a gloomy and mysterious place, and almost immediately, Flavia discovers a mummified body in the chimney of her room. If you have not made Flavia's acquaintance before, she is definitely worth discovering. She has been compared to a cross between Harriet the Spy and Sherlock Holmes, and that is an apt description. If you are new to the series, it's probably best to begin with The Sweetness at the bottom of the Pie, which is the first of Flavia's adventures.
Janet Evanovich latest book Top Secret Twenty One has Stephanie Plum at it again. Stephanine Plum is a bounty hunter but not what you would think of as a typical bounty hunter. She gets by on being cute and lots of luck. In Top Secret Twenty One she is looking for Jimmy Poletti who is out on bail and has missed his court date. While hunting him down she keeps finding his poker buddies, one after the other, dead. She is also helping Ranger who has an old Russian enemy trying to kill him. As usual, Stephanie bumbles about and defies death a few times. You know how some hotels have a parking garage attached and there is a walkway from the hotel to the garage, well Stephanie finds herself on top of this walkway with a bad guy forcing her to cross it. Mix in 10 Chihuahuas and you have yourself another Stephanie Plum humorous bounty hunt. I have been reading Janet Evanovich for years, this is my first time listening to one of her books on audio downloaded from KPL's overdrive. I love audio books but since I have been reading about Stephanie, Lulu, Ranger etc I have these characters in my head. Hearing a voice for Lulu that was not what my head envisioned bothered me. I and others had similar issues when the movie One for the Money came out. It was a good movie but each of us had preconceived impressions of how these people looked and talked. I think this is the highest praise we can give an author, Janet Evanovich has written her books so well that we have made these fictional characters real in our heads. I love the Stephanie Plum series although I keep thinking my goodness how inept can you be.Come on down to KPL and check it out.
Invisible by James Patterson. Emmy Dockery is a research assistant for the FBI and thinks she has discovered a serial killer who uses arson to cover his tracks. The problem for Emmy is that the killer is very good at making these house fires look accidental. Also unfortunately for Emmy one of the victims was her twin sister so everyone discounts her theory thinking she is just mourning her sister. She enlists the aid of her ex finance, who is also ex FBI to look into the fires. They eventually find evidence and the hunt is on. The story is eerie and you feel Emmy's frustrations. The author also lets us hear from the killer. The killer narrates the Graham Chronicles where we hear what the killer is thinking and what happens to a victim. This lets us the reader be on both sides on story, we watch Emmy track and try and hunt the killer down, we also hear from the killer and we are shared insight into the killers thoughts and actions. I downloaded this book from KPL as an mp3 and listened to the story. When the killer was talking, it sounded very pompously and full of self-importance. I took an instant dislike to the killer, but to be fair I wasn't all that crazy about Emmy. Emmy was not a team player, she ignored Book's commands and just did whatever she wanted even if that meant she might have endangered fellow agents. I was glad when Bookman finally got fed up with her and denounced her. That was almost more satisfying than them catching the killer. Come on down to KPL and check it out.
Scandanavian crime noir seems to be recognized as its own genre, and this new title by Camilla Lackberg certainly falls into that category. This is the seventh in the series that began with The Ice Princess, and features police detective Patrick Hedberg and true crime writer Erica Falck, now married and expecting their first child. Erica finds a Nazi war medal and diaries among her deceased mother's possessions, and goes to visit one of her mother's old friends, a historian and expert on World War 2, for more information. The man is brutally murdered soon after Erica's visit. Woven into the crime novel are scenes of everyday life in the small Swedish town of Fjallbacka, and characters that are interesting in their own right. Some reviews found this bothersome, but I didn't. Even police detectives and true crime authors have real lives, running parallel to their professional ones. If you haven't yet discovered author Lackberg, it's probably best to begin with her first book , The Ice Princess, although this novel can certainly stand on its own. Full of well placed clues and historical fact from the 1940's, this kept me guessing until the end.
I watched the television show "Castle". In the television show Richard Castle is a writer who gets to ride along with Detective Kate Becket and her team. In the television show (and I emphasize this) Richard Castle (played by Nathan Fillion) writes a book about Detective Kate Becket and calls her Nikki Heat. Someone thought hey lets write a real book about Nikki Heat and pretend it is written by Richard Castle just like in the television series. There are now 6 books in the Nikki Heat Series written by Richard Castle. Nobody knows who really writes these books. The book jacket shows a picture of Nathan Fillion but they say his name is Richard Castle. Nathan Fillion has even signed some books using his television name Richard Castle. The books have the same characters as in the show but they change the names. Richard Castle is called Jameson Rook, Kate Becket is of course Nikki Heat, Detective Ryan is Raley and Detective Esposito is Ochoa. I like that they renamed Castle as Rook. In Chess in a move called Castling, the Rook can change positions with the King. There after you call that piece a Castle. I like that they choose to use that play on words. When reading the books it is like watching the show but can get confusing. I was reading one of the books and watching the show at the same time, not exactly the same same time, and was getting a wee bit confused. In the show her mother was killed in an alley, but in the book she is killed in their kitchen. On the show during think tank sessions they toss around a little ball the size of a tennis ball, in the book they toss a basketball, I like the show version better. I downloaded from KPL and listened to these books on my mp3 player, KPL also has the print versions and digital. So whether you prefer print, digital or audio KPL has it all. Check it out at KPL.
I'll admit flat out that I'm a huge fan of author Kate Atkinson. In her fourth novel featuring semiretired detective Jackson Brodie, "Started Early, Took my Dog", the author delves into the subject of missing children. Jackson is searching for the biological parents of an Australian woman; it seems straightforward at first, but soon he has more questions than answers. He is also dealing with his teenage daughter by his first wife, his former lover and their son, and a dog that he impulsively rescues from an abusive owner. Concurrently, recently retired detective Tracy Waterhouse, lonely and somewhat jaded after seeing the darker side of life for decades, sees a young girl being dragged along by a prostitute, and something snaps- Tracy offers the woman cash for the kid, and suddenly she is a parent of a child in a fairy costume. Bad people are soon pursuing them, but they don't seem to want the child back. All roads lead to Jackson; it emerges that he and Tracy are working towards the same end in solving their separate mysteries. Kate Atkinson takes these potentially dark events and injects them with her sharp observations and wit. Previous novels in the Jackson Brodie series are equally great reading, and her best selling novel, "Life After Life" has won several awards, including the COSTA award in 2013.
The hazmat killer's recent victim is found on a carousel and Zach and Kylie of the elite NYPD Red force must find him before the mayors re-election vote. The hazmat killer is killing people that the legal system was unable to bring to justice. They torture the bad guy and video tape the confession, kill him and then leave the body in a very public place. The video is then released to the internet. Kylie and Zach have a hard time getting people to help as most are routing for the vigilante. NYPD RED 2 is James Patterson and Marshall Karp's second book in this series. While you can read this book without having first read NYPD RED, I recommend reading them in order. Kylie and Zach have a romantic history and it's just better if you read about it in the first book as they talk about it a lot (way more than I wanted) in the second book.You can find both books at KPL, as well as thousand of others both in hard print and digital.
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I am a practicing dentist and I am considering the guard for student loan repayment. I have not gotten a direct answer from recruiters but I have federal and private student loans. Will my private loans be eligible for repayment? Anyone had this situation?
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Re: Dentist HPLRP
Originally posted by Eagle01 View PostI am a practicing dentist and I am considering the guard for student loan repayment. I have not gotten a direct answer from recruiters but I have federal and private student loans. Will my private loans be eligible for repayment? Anyone had this situation?
No, your private loans will not qualify for SLRP. See the SLRP sticky in the Benefits section for more information.
Super Moderator
- May 2006
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Lieutenant Colonel, Virginia ARNG
The answer to your question may already be here.
Re: Dentist HPLRP
I trust that you're working with an AMEDD recruiter. No one else can even pretend to deal with medical incentives.
Re: Dentist HPLRP
I am working with a recruiter but I am in the early stages of getting him all of my credentials and he doesn't have a clear answer either. It seems that SLRP is different than HPLRP. I don't know many post-graduates that come out of school without private education loans considering $18,500 is the max federal loan you are allowed per year. That doesn't even cover the cost of tuition.
Re: Dentist HPLRP
Ditto what LTC Ritchie says. Recruiters will give lots of well-meaning advice, but AMEDD benefits and policies are worlds away from non_AMEDD. For instance, disregard the link to SLRP. HPLRP is completely different.
That said, OP, private loans DO NOT qualify for HPLRP. I know you mentioned there being a $18K year federal loan limit, but this isn't true. $18K (I think it's $18.5K?) is the limit for federally SUBSIDIZED loans, but you can qualify for an additional $38K in federally UNSUBSIDIZED loans. Might you be confusing subsidized vs. unsubsidized with private vs. government? I know this to be true, because I am looking down the barrel at close to $200K in federal loans for four years of medical school.
As a licensed dentist, you qualify for $40K per year for a maximum of $240K total towards your student loans, but only for your federal loans. Your private loans do not qualify. The policy is very explicit about this. One of my roles in the Guard while in medical school was AMEDD Officer Recruiting.
By the way, you might focus your questions on one thread rather than multiple ones. It cuts down on the efforts of folks trying to help you.


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|Publication number
|Publication type
|Application number
|Publication date
|Mar 10, 2011
|Filing date
|Nov 8, 2010
|Priority date
|Jan 22, 2003
|Also published as
|Publication number
|John A. Moon, Alan D. Kersey, Martin A. Putnam, Tuo Li
|Original Assignee
|Illumina, Inc.
|Export Citation
|BiBTeX, EndNote, RefMan
|Referenced by (6), Classifications (50), Legal Events (2)
|External Links: USPTO, USPTO Assignment, Espacenet
This application is a continuation of U.S. application Ser. No. 12/644,255, filed Dec. 22, 2009, which is a continuation of U.S. application Ser. No. 11/607,837 (the '837 application), filed Nov. 30, 2006, entitled "Hybrid Random Bead/Chip Base Microarray." The '837 application is a continuation of U.S. application Ser. No. 10/763,995 (the '995 application) (issued as U.S. Pat. No. 7,164,533), filed Jan. 22, 2004, which claims the benefit of U.S. Provisional Patent Applications Nos. 60/441,678, filed Jan. 22, 2003, entitled "Hybrid Random Bead/Chip Microarray"; and 60/519,932, filed Nov. 14, 2003, entitled, "Diffraction Grating-Based Encoded Microparticles for Multiplexed Experiments." The '995 application also claims priority to and is a continuation-in-part of U.S. patent application Ser. Nos. 10/661,234, filed Sep. 12, 2003, entitled "Diffraction Grating-Based Optical Identification Element"; 10/661,031, filed Sep. 12, 2003, entitled "Diffraction Grating-Based Encoded Micro-Particles for Multiplexed Experiments"; and 10/661,836, filed Sep. 12, 2003, entitled "Method and Apparatus for Aligning Microbeads in order to Interrogate the Same". All of the foregoing applications are incorporated herein by reference in their entirety.
U.S. patent application Ser. Nos. 10/661,082, 10/661,115, 10/661,254 and 10/661,116, filed Sep. 12, 2003, contain subject matter related to that disclosed herein, all of which are incorporated by reference in their entirety.
This invention relates to optical identification, and more particularly to diffraction grating-based encoded optical elements/micro-particles for performing multiplexed experiments.
A common class of experiments, known as a multiplexed assay or multiplexed experiment, comprises mixing (or reacting) a labeled target analyte or sample (which may have known or unknown properties or sequences) with a set of "probe" or reference substances (which also may have known or unknown properties or sequences). Multiplexing allows many properties of the target analyte to be probed or evaluated simultaneously (i.e., in parallel). For example, in a gene expression assay, the "target" analyte, usually an unknown sequence of DNA, is labeled with a fluorescent molecule to form the labeled analyte.
In a known DNA/genomic sequencing assay, each probe consists of known DNA sequences of a predetermined length, which are attached to a labeled (or encoded) bead or to a known location or position (or spot) on a substrate.
When the labeled target analyte is mixed with the probes, segments of the DNA sequence of the labeled target analyte will selectively bind to complementary segments of the DNA sequence of the known probe. The known probes are then spatially separated and examined for fluorescence. The probes that fluoresce indicate that the DNA sequence strands of the target analyte have attached or hybridized to the complementary DNA of the probe. The DNA sequences in the target analyte can then be determined by knowing the complementary DNA (or cDNA) sequence of each known probe to which the labeled target is attached. In addition the level of fluorescence is indicative of how many target molecules hybridized to the probe molecules for a given bead or spot on a substrate.
Generally, the probes are identified either by spatial location on a substrate or by attaching the probe to a bead or particle that is labeled (or encoded) to identify the probe, and ultimately the "target" analyte. The first approach separates the probes in a predetermined grid, where the probe's identity is linked to its position on the grid. One example of this is a "chip" format, where DNA is attached to a 2-D substrate or microarray, where oligomer DNA sequences are selectively attached (either by spotting or grown) onto small sections or spots on the surface of the substrate in a predetermined spatial order and location on a substrate (usually a planar substrate, such as a glass microscope slide), such as that sold by Affymetrix and others.
A second or "bead-based" approach, for identifying the probe allows the probes to mix without any specific spatial position, which is often called the "random bead assay" approach. In this approach the probes are attached to a small bead or particle instead of a larger substrate so they are free to move (usually in a liquid medium). This approach has an advantage in that the analyte reaction can be performed in a liquid/solution by conventional wet-chemistry techniques, which gives the probes a better opportunity to interact with the analyte. However, this approach requires that each bead or probe be individually identifiable.
There are many known methods and substrate types that can be used for tagging or otherwise uniquely identifying individual beads with attached probes. Known methods include using polystyrene latex spheres that are colored or fluorescent labeled, such as that sold by Luminex and others. Other methods include using small plastic cans with a conventional bar code applied, or a small container includes a solid support material and a radio-frequency tag, such as that sold by Pharmaseq and others.
The beads have the advantage of using liquid or solution based chemistry and flexibility but current bead technology does have a limited number of identifiable codes and/or are not suitable for harsh environments/chemicals. Whereas chips typically have the advantage of having higher density (or high multiplexing) capability than beads and can be read using standard fluorescence scanners, but are not as flexible or economically customizable as beads.
Therefore, it would be desirable to provide a platform with benefits of both the bead-based platforms and the chip-based platforms.
Objects of the present invention include provision of a platform that provides benefits of both bead-based platforms and chip-based platforms.
According to the present invention, a method of performing an assay process is provided comprising the steps of: providing microbeads in a solution, each microbead having a particle substrate with a grating with a superposition of different predetermined regular periodic variations of the index of refraction disposed in the particle along a grating axis and indicative of a code; placing the microbeads on an alignment substrate; reading codes of the microbeads and the position thereof on the alignment substrate; reading the fluorescence on each microbead and the position thereof on the alignment substrate; and determining an assay result based on bead position and bead code of the earlier reading steps. The particle substrate may be formed of a transparent dielectric material with the index of refraction at each point in the dielectric material, and the superposition of different regular periodic variations in the index of refraction is disposed along the length of the particle substrate.
The present invention also includes apparatus for reading microbeads that form part of an assay process, comprising: an alignment substrate for receiving the microbeads thereon: and a bead mapper for reading codes of the microbeads and the position thereof on the alignment substrate.
The invention is a significant improvement over chip-based assay platforms and existing bead-based assay platforms. In particular, the bead assay can be performed with solution or wet chemistry, then when the experiment is completed, the beads are placed on a slide, plate, or substrate (e.g., a groove plate) which aligns the beads. The beads are then placed in a "bead mapper", which reads the codes and maps each bead code with a unique position on the slide. Once the beads have been mapped, the slide may be placed in any standard scanner capable of detecting the label used for the analyte and its position on the slide. For example, a standard fluorescence reader/scanner used to read chip-based microarrays may be used to read the fluorescence intensity at each bead location on the slide, similar to reading the fluorescence of each spot on the chip. The intensity/location information is then combined with the code/location information to determine which probes are exhibiting fluorescence, and the intensity thereof.
The invention may be viewed as a "chip" or "microchip" approach where the probes (or beads) are assembled from many individually fabricated parts. The beads may be ordered in one dimension along the grooves, but are randomly distributed (but oriented) along each groove. However, any technique may be used that allows the bead location to be identified.
This self-assembled "chip" approach has many advantages over conventional bead based assays. In particular, since the beads are fixed on a chip substrate (e.g., groove plate), they may be examined and re-examined at any time. Also, beads of interest can be easily removed and sorted from the plate/chip after an experiment is performed. More specifically, after reading the chip, the beads may be removed from the chip for further and/or alternative processing or experiments. If desired, the chip substrate and/or the beads may be reused in other experiments or assays. Further, a fixed plate format is easier to use in experiments that vary the temperature. Still further, a fixed plate format allows convenient use of a standard chip reader to examine the beads. Also, the beads do not need to be examined using a flow cytometer.
Alternatively, instead of performing the analyte reaction or hybridization reaction before placing the beads on the chip, the beads (or probe particles) can be assembled into the chip format before the analyte reaction process. In that case, the analyte can be applied to the chip with the beads disposed thereon, in which case the analyte reaction would occur on the chip.
The microbeads are inexpensive to manufacture and the identification codes are easy and inexpensive to imprint into the microbeads. The codes are digitally readable and easily adapted to optical coding techniques. Thus, the bead mapper optical readout is very simple and inexpensive to implement. Further, the invention allows for the use of a standard scanner to the label used for the analyte, which may avoid the need to purchase an additional scanner.
Further, the beads may be oriented in 1-D in grooves (which may or may not be linear) and are randomly distributed along the grooves. Also, the beads need not be fixed in any way in the grooves other than by capillary force if desired.
The code on the bead is not affected by spot imperfections, scratches, cracks or breaks. In addition, splitting or slicing an element axially produces more elements with the same code; therefore, when a bead is axially split-up, the code is not lost, but instead replicated in each piece. Unlike electronic ID elements, the elements of the present invention are not affected by nuclear or electromagnetic radiation.
The invention may be used in any assay or multiplexed experiment. The present invention may be used with any known combinatorial chemistry or biochemistry assay process, and are especially adaptable to assays having solid phase immobilization. The invention may be used in many areas such as drug discovery, functionalized substrates, biology, proteomics, combinatorial chemistry, and any assays or multiplexed experiments. Examples of common assays are SNP (single nucleotide polymorphism) detection, DNA/genomic sequence analysis, genotyping, gene expression assays, proteomics assay, peptide assays, antigen/antibody assays (immunoassay), ligand/receptor assays, DNA analysis/tracking/sorting/tagging, as well as tagging of molecules, biological particles, cell identification and sorting, matrix support materials, receptor binding assays, scintillation proximity assays, radioactive or non-radioactive proximity assays, and other assays, high throughput drug/genome screening, and/or massively parallel assay applications. The analyte can be labeled, detected or identified with any technique capable of being used in an assay with arrays or beads, including but not limited to fluorescent, luminescent, phosphorescent, quantum dot, light scattering colloidal particles, radioactive isotopes, mass spectroscopy, NMR (nuclear magnetic resonance), EPR (electro paramagnetic resonance), ESR (electron spin resonance), IR (infrared), FTIR (Fourier transform infra red), Raman spectroscopy, or other magenetic, vibrational, electromagnetic, or optical labeling or detection techniques. Accordingly, the scanner may any scanner capable of measuring or sensing any of the foregoing analyte labels.
The invention provides uniquely identifiable beads with reaction supports by active coatings for reaction tracking to perform multiplexed experiments. The invention may also be used in any chemical and/or biochemical purification, isolation, or filtering-type process where bead or bead-like solid supports may be used (e.g., chromatographic techniques, such as affinity column purification). In that case, the above techniques for labeling, detection or identification may be used.
The foregoing and other objects, features and advantages of the present invention will become more apparent in light of the following detailed description of exemplary embodiments thereof.
Referring to
The optical identification element 8 described herein is similar to that described in Copending U.S. patent application Ser. No. 10/661,234, filed Sep. 12, 2003, entitled "Diffraction Grating-Based Optical Identification Element", which is incorporated herein by reference in its entirety.
In particular, the substrate 10 has an inner region 20 where the grating 12 is located. The inner region 20 may be photosensitive to allow the writing or impressing of the grating 12. The substrate 10 has an outer region 18, which does not have the grating 12 therein.
The grating 12 is a combination of one or more individual spatial periodic sinusoidal variations (or components) in the refractive index that are collocated at substantially the same location on the substrate 10 along the length of the grating region 20, each having a spatial period (or pitch) Λ. The resultant combination of these individual pitches is the grating 12, comprising spatial periods (Λ1-Λn) each representing a bit in the code. Thus, the grating 12 represents a unique optically readable code, made up of bits, where a bit corresponds to a unique pitch Λ within the grating 12. Accordingly, for a digital binary (0-1) code, the code is determined by which spatial periods (Λ1-Λn) exist (or do not exist) in a given composite grating 12. The code or bits may also be determined by additional parameters (or additional degrees of multiplexing), and other numerical bases for the code may be used, as discussed herein and/or in the aforementioned patent application.
The grating 12 may also be referred to herein as a composite or collocated grating. Also, the grating 12 may be referred to as a "hologram", as the grating 12 transforms, translates, or filters an input optical signal to a predetermined desired optical output pattern or signal.
The substrate 10 has an outer diameter D1 and comprises silica glass (SiO2) having the appropriate chemical composition to allow the grating 12 to be disposed therein or thereon. Other materials for the optical substrate 10 may be used if desired. For example, the substrate 10 may be made of any glass, e.g., silica, phosphate glass, borosilicate glass, or other glasses, or made of glass and plastic, or solely plastic. For high temperature or harsh chemical applications, the optical substrate 10 made of a glass material is desirable. If a flexible substrate is needed, plastic, rubber or polymer-based substrate may be used. The optical substrate 10 may be any material capable of having the grating 12 disposed in the grating region 20 and that allows light to pass through it to allow the code to be optically read.
The optical substrate 10 with the grating 12 has a length L and an outer diameter D1, and the inner region 20 diameter D. The length L can range from very small "microbeads" (or microelements, micro-particles, or encoded particles), about 1-1000 microns or smaller, to larger "macroelements" for larger applications (about 1.0-1000 mm or greater). In addition, the outer dimension D1 can range from small (less than 1000 microns) to large (1.0-1000 mm and greater). Other dimensions and lengths for the substrate 10 and the grating 12 may be used.
The grating 12 may have a length Lg of about the length L of the substrate 10. Alternatively, the length Lg of the grating 12 may be shorter than the total length L of the substrate 10.
The outer region 18 is made of pure silica (SiO2) and has a refractive index n2 of about 1.458 (at a wavelength of about 1553 nm), and the inner grating region 20 of the substrate 10 has dopants, such as germanium and/or boron, to provide a refractive index n1 of about 1.453, which is less than that of outer region 18 by about 0.005. Other indices of refraction n1,n2 for the grating region 20 and the outer region 18, respectively, may be used, if desired, provided the grating 12 can be impressed in the desired grating region 20. For example, the grating region 20 may have an index of refraction that is larger than that of the outer region 18 or grating region 20 may have the same index of refraction as the outer region 18 if desired.
Referring to
The detector 29 has the necessary optics, electronics, software and/or firmware to perform the functions described herein. In particular, the detector reads the optical signal 27 diffracted or reflected from the grating 12 and determines the code based on the pitches present or the optical pattern, as discussed more herein or in the aforementioned patent application. An output signal indicative of the code is provided on a line 31.
Referring to
Referring to
Next, in step 46, the chip is provided to a Reader/Scanner 824 (
Accordingly, as discussed hereinabove, the assay utilizes the fact that each probe particle (or microbead) is individually identifiable. Once the bead identification code or tag is read, and the spatial position (or location) is known, the self-assembled "chip" can be inserted into a conventional known chip reader or scanner 824 (
Examples of known chip readers include the following: Axon Gene Pix Pro 4100 A, GSI/Lumonics/Perkin Elmer Scanner, Alpha Inatech, and others. Other commercial readers or scanners now known or later developed may be used provided it can detect the desired analyte reaction parameter, e.g., fluorescence, etc., and the it can provide the location of same on the substrate.
Alternatively, the reader/scanner 824 may be similar to the analyte reaction reading and analysis portions of the microbead reader device described in Copending Provisional Patent Application Ser. No. 60/512,302 (Docket No. CC-0667PR), entitled "Optical Reader for Diffraction Grating Based Encoded Microbeads", filed Oct. 17, 2003; Ser. No. 60/513,053 (Docket No. CC-0669PR), filed Oct. 21, 2003, "Optical Reader for Diffraction Grating Based Encoded Microbeads"; Ser. No. 60/508,038 (Docket No. CC-0577PR), "Optical Reader for Diffraction Grating Based Encoded Microbeads", filed Oct. 1, 2003, all of which are incorporated herein by reference in their entirety.
Similarly, the Bead Mapper 20 may be similar to the bead reading/mapping portions of the microbead reader described in Copending Provisional Patent Application Ser. No. 60/512,302 (Docket No. CC-0667PR), entitled "Optical Reader for Diffraction Grating Based Encoded Microbeads", filed Oct. 17, 2003; Ser. No. 60/513,053 (Docket No. CC-0669PR), filed Oct. 21, 2003, "Optical Reader for Diffraction Grating Based Encoded Microbeads"; Ser. No. 60/508,038 (Docket No. CC-0577PR), "Optical Reader for Diffraction Grating Based Encoded Microbeads", filed Oct. 1, 2003, all of which are incorporated herein by reference in their entirety.
Referring to
For example, DNA probe molecules may be directly synthesized on the beads using standard phosphoramidite chemistry with no post synthetic purification, and the beads used as the solid support. The attachment to the bead may be done by preparing the beads using standard linker chemistry coated on the beads that allows the probe to attach to the bead. Then, the oligo probe may be grown base-by-base to create the oligo sequence. Alternatively, the entire desired oligo sequence may be pre-fabricated and then attached to the bead after fabrication. In that case, the linker chemistry used on the bead would likely be different and possibly more complex than the linker chemistry used in direct synthesis. Also, the beads may be functionalized as discussed hereinbefore and then placed in a blocker solution of BSA Bovine Serum Albumin (or any other suitable blocker to prevent non-specific binding of the target molecule). The beads may then be hybridized by placing the beads in a hybridization solution. Any desirable hybridization solution may be used. One example is: 5� concentration of SSC (Standard Saline Citrate), 25% formamide, 0.1% SDS (Sodium Dodecyl Sulfate-soap—used to help the beads not stick to the walls of tube), a predetermined amount of complementary DNA (cDNA) to the sequence of a given Probe tagged with Cy3 fluorescent molecules, and a predetermined amount of complementary DNA (cDNA) to the sequence of that Probe tagged with Cy5 fluorescent molecules. Any other hybridization or analyte reaction technique may be used if desired.
Referring to
The "target" analytes 52-54 within the solution 60 are then mixed with the functionalized microbeads 72-74. During the mixing of the "target" analytes 52-54 and the functionalized microbeads 72-74, the "target" analytes attach to the complementary probes 76-78, as shown for functionalized microbeads 72,73 having codes 12345678 and 34128913. Specifically, as shown in
For example as discussed hereinbefore, each coded functionalized microbead 72-74 has a unique probe 76-78, respectively bonded thereto, such as a portion of a single strand of DNA. Similarly, the "target" analytes 52-54 comprise a plurality of unknown and unique single strands of DNA. These "target" analytes 52-54 are also processed with a fluorescent, such as dyeing, such that the test molecules illuminate. As will be discussed hereinafter, the fluorescence of the "target" analytes provide the means to identify, which functionalized microbeads 72-74 have a "target" analyte attached thereto.
Once the reaction or combining or hybridization is complete, the functionalized (or reacted or hybridized) microbeads 72-74 are rinsed off with a saline solution to clean off the uncombined "target" analytes 52-54.
Referring to
Referring to
Referring to
Referring to
Referring to
The tray 84 is made of glass or plastic or any material that is transparent to the code reading incident beam 24 and code reading output light beams 27 as well as the fluorescent excitation beam 800 and the output fluorescent optical signal 802, and is properly suited for the desired application or experiment, e.g., temperature range, harsh chemicals, or other application specific requirements.
The code signal 822 from the bead code reader 820 and the fluorescent signal 810 from the fluorescence detector are provided to a known computer 812. The computer reads the code associated with each bead and determines the chemical probe that was attached thereto from a predetermined table that correlates a predetermined relationship between the bead code and the attached probes. In addition, the computer 812 reads the fluorescence associated with each bead and determines the sample or analyte that is attached to the bead from a predetermined data that correlates a predetermined relationship between the fluorescence tag and the analyte attached thereto. The computer 812 then determines information about the analyte and/or the probe as well as about the bonding of the analyte to the probe, and provides such information on a display, printout, storage medium or other interface to an operator, scientist or database for review and/or analysis, as indicated by a line 815.
Generally, the assay of the present invention may be used to carry out any binding assay or screen involving immobilization of one of the binding agents. Such solid-phase assays or screens are well known in the chemical and biochemical arts. For example, such screening may involve specific binding of cells to a molecule (e.g. an antibody or antigen) immobilized on a microbead in the assay followed by analysis to detect whether or to what extent binding occurs. Alternatively, the beads may subsequently removed from the groove plate for sorting and analysis via flow cytometry (see e.g. by Needels et al, Proc. Natl. Acad. Sci. USA, Vol. 90, pp. 10700-10704, November 1993). Examples of biological compounds that may be assayed or screened using the assay of the present invention include, e.g. agonists and antagonists for cell membrane receptors, toxins, venoms, viral epitopes, hormones, sugars, cofactors, peptides, enzyme substrates, drugs inclusive of opiates and steroids, proteins including antibodies, monoclonal antibodies, antisera reactive with specific antigenic determinants, nucleic acids, lectins, polysaccharides, cellular membranes and organelles. In addition, the present invention may be used in any of a large number of well-known hybridization assays where nucleic acids are immobilized on a surface of a substrate, e.g. genotyping, polymorphism detection, gene expression analysis, fingerprinting, and other methods of DNA- or RNA-based sample analysis or diagnosis.
Any of the great number of isotopic and non-isotopic labeling and detection methods well-known in the chemical and biochemical assay art may be used to detect binding with the present invention. Alternatively, spectroscopic methods well-known in the art may be used to determine directly whether a molecule is bound to a surface coating in a desired configuration. Spectroscopic methods include e.g., UV-VIS, NMR, EPR, IR, Raman, mass spectrometry and other methods well-known in the art. For example, mass spectrometry also is now widely employed for the analysis of biological macromolecules. The method typically involves immobilization of a protein on a surface of substrate where it is then exposed to a ligand binding interaction. Following ligand binding (or non-binding) the molecule is desorbed from the surface and into a spectrometer using a laser (see, e.g. Merchant and Weinberger, "Recent advancements in surface-enhanced laser desorption/ionization-time of flight-mass spectrometry," Electrophoresis 21:1164-1177 (2000)). The microbeads in the assay of the present invention may be used as substrates in the mass spectrometry detection methods described above.
Various aspects of the present invention may be conducted in an automated or semi-automated manner, generally with the assistance of well-known data processing methods. Computer programs and other data processing methods well known in the art may be used to store information including e.g. microbead identifiers, probe sequence information, sample information, and binding signal intensities. Data processing methods well known in the art may be used to read input data covering the desired characteristics.
The invention may be used in many areas such as drug discovery, functionalized substrates, biology, proteomics, combinatorial chemistry, DNA analysis/tracking/sorting/tagging, as well as tagging of molecules, biological particles, matrix support materials, immunoassays, receptor binding assays, scintillation proximity assays, radioactive or non-radioactive proximity assays, and other assays, (including fluorescent, mass spectroscopy), high throughput drug/genome screening, and/or massively parallel assay applications. The invention provides uniquely identifiable beads with reaction supports by active coatings for reaction tracking to perform multiplexed experiments.
Some current techniques used in combinatorial chemistry or biochemistry are described in U.S. Pat. No. 6,294,327, entitled "Apparatus and Method for Detecting Samples Labeled With Material Having Strong Light Scattering Properties, Using Reflection Mode Light and Diffuse Scattering", issued Sep. 23, 2001 to Walton et al.; U.S. Pat. No. 6,242,180, entitled "Computer Aided Visualization and Analysis System for Sequence Evaluation", issued Jun. 5, 2001, to Chee; U.S. Pat. No. 6,309,823 entitled "Arrays of Nucleic Acid Probes for Analyzing Biotransformation of Genes and Methods of Using the Same", Oct. 30, 2001, to Cronin et al.; U.S. Pat. No. 6,440,667, entitled "Analysis of Target Molecules Using an Encoding System"; U.S. Pat. No. 6,355,432, entitled "Products for Detecting Nucleic Acids"; U.S. Pat. No. 6,197,506, entitled "Method of Detecting Nucleic Acids"; U.S. Pat. No. 6,309,822, entitled "Method for comparing copy number of nucleic acid sequences"; U.S. Pat. No. 5,547,839, entitled "Sequencing of surface immobilized polymers utilizing micro-fluorescence detection", U.S. Pat. No. 6,383,754, entitled "Binary Encoded Sequence Tags", and U.S. Pat. Nos. 6,261,782 and 6,667,121, entitled "Fixed Address Analysis of Sequence Tags", which are all incorporated herein by reference to the extent needed to understand the present invention.
The invention can be used in combinatorial chemistry, active coating and functionalized polymers, as well as immunoassays, and hybridization reactions. The invention enables millions of parallel chemical reactions, enable large-scale repeated chemical reactions, increase productivity and reduce time-to-market for drug and other material development industries.
As discussed hereinbefore, although a fluorescent label is probably most convenient, other sorts of labels, e.g., radioactive, enzyme linked, optically detectable, or spectroscopic labels may be used. An appropriate detection method applicable to the selected labeling method can be selected. Suitable labels include radionucleotides, enzymes, substrates, cofactors, inhibitors, magnetic particles, heavy metal atoms, and particularly fluorescers, chemiluminescers, and spectroscopic labels. Patents teaching the use of such labels include U.S. Pat. Nos. 3,817,837; 3,850,752; 3,939,350; 3,996,345; 4,277,437; 4,275,149; and 4,366,241.
With an appropriate label selected, the detection system best adapted for high resolution and high sensitivity detection may be selected. As indicated above, an optically detectable system, e.g., fluorescence or chemilumnescence would be preferred but is not required. Other detection systems may be adapted to the purpose, e.g., electron microscopy, scanning electron microscopy (SEM), scanning tunneling electron microscopy (STEM), infrared microscopy, atomic force microscopy (AFM), electrical conductance, and image plate transfer.
Referring to
Referring to
Each of the individual spatial periods (Λ1-Λn) in the grating 12 is slightly different, thus producing an array of N unique diffraction conditions (or diffraction angles) discussed more hereinafter. When the element 8 is illuminated from the side, in the region of the grating 12, at an appropriate input angle, e.g., about 30 degrees, with a single input wavelength λ (monochromatic) source, the diffracted (or reflected) beams 26-36 are generated. Other input angles θi may be used if desired, depending on various design parameters as discussed herein and/or in the aforementioned patent application, and provided that a known diffraction equation (Eq. 1 below) is satisfied:
sin(θi)+sin(θo)=mλ/nΛ Eq. 1
where Eq. 1 is diffraction (or reflection or scatter) relationship between input wavelength λ, input incident angle θi, output incident angle θo, and the spatial period Λ of the grating 12. Further, m is the "order" of the reflection being observed, and n is the refractive index of the substrate 10. The value of m=1 or first order reflection is acceptable for illustrative purposes. Eq. 1 applies to light incident on outer surfaces of the substrate 10 which are parallel to the longitudinal axis of the grating (or the kB vector). Because the angles θi,θo are defined outside the substrate 10 and because the effective refractive index of the substrate 10 is substantially a common value, the value of n in Eq. 1 cancels out of this equation.
Thus, for a given input wavelength λ, grating spacing Λ, and incident angle of the input light θi, the angle θo of the reflected output light may be determined. Solving Eq. 1 for θo and plugging in m=1, gives:
θo=sin−1(λ/Λ−sin(θi)) Eq. 2
For example, for an input wavelength λ=532 nm, a grating spacing Λ=0.532 microns (or 532 nm), and an input angle of incidence θi=30 degrees, the output angle of reflection will be θo=30 degrees. Alternatively, for an input wavelength λ=632 nm, a grating spacing Λ=0.532 microns (or 532 nm), and an input angle θi of 30 degrees, the output angle of reflection θo will be at 43.47 degrees, or for an input angle θi=37 degrees, the output angle of reflection will be θo=37 degrees. Any input angle that satisfies the design requirements discussed herein and/or in the aforementioned patent application may be used.
In addition, to have sufficient optical output power and signal to noise ratio, the output light 27 should fall within an acceptable portion of the Bragg envelope (or normalized reflection efficiency envelope) curve 200, as indicated by points 204,206, also defined as a Bragg envelope angle θB, as also discussed herein and/or in the aforementioned patent application. The curve 200 may be defined as:
where K=2πδn/λ, where, δn is the local refractive index modulation amplitude of the grating and λ is the input wavelength, sin c(x)=sin(x)/x, and the vectors ki=2π cos(θi)/λ and ko=2π cos(θo)/λ are the projections of the incident light and the output (or reflected) light, respectively, onto the line 203 normal to the axial direction of the grating 12 (or the grating vector kB), D is the thickness or depth of the grating 12 as measured along the line 203 (normal to the axial direction of the grating 12). Other substrate shapes than a cylinder may be used and will exhibit a similar peaked characteristic of the Bragg envelope. We have found that a value for δn of about 10−4 in the grating region of the substrate is acceptable; however, other values may be used if desired.
Rewriting Eq. 3 gives the reflection efficiency profile of the Bragg envelope as:
where: x=(ki−ko)D/2=(πD/λ)*(cos θi−cos θo)
Thus, when the input angle θi is equal to the output (or reflected) angle θo (i.e., θi=θo, the reflection efficiency I (Eqs. 3 & 4) is maximized, which is at the center or peak of the Bragg envelope. When θi=θo, the input light angle is referred to as the Bragg angle as is known. The efficiency decreases for other input and output angles (i.e., θi≠θo), as defined by Eqs. 3 & 4. Thus, for maximum reflection efficiency and thus output light power, for a given grating pitch Λ and input wavelength, the angle θi of the input light 24 should be set so that the angle θo of the reflected output light equals the input angle θi.
Also, as the thickness or diameter D of the grating decreases, the width of the sin(x)/x function (and thus the width of the Bragg envelope) increases and, the coefficient to or amplitude of the sin c2 (or (sin(x)/x)2 function (and thus the efficiency level across the Bragg envelope) also increases, and vice versa. Further, as the wavelength λ increases, the half-width of the Bragg envelope as well as the efficiency level across the Bragg envelope both decrease. Thus, there is a trade-off between the brightness of an individual bit and the number of bits available under the Bragg envelope. Ideally, δn should be made as large as possible to maximize the brightness, which allows D to be made smaller.
From Eq. 3 and 4, the half-angle of the Bragg envelope θB is defined as:
where η is a reflection efficiency factor which is the value for x in the sin c2(x) function where the value of sin c2(x) has decreased to a predetermined value from the maximum amplitude as indicated by points 204,206 on the curve 200.
We have found that the reflection efficiency is acceptable when η≦1.39. This value for η corresponds to when the amplitude of the reflected beam (i.e., from the sin c2(x) function of Eqs. 3 & 4) has decayed to about 50% of its peak value. In particular, when x=1.39=η, sin c2(x)=0.5. However, other values for efficiency thresholds or factor in the Bragg envelope may be used if desired.
The beams 26-36 are imaged onto the CCD camera 60 to produce the pattern of light and dark regions 120-132 representing a digital (or binary) code, where light=1 and dark=0 (or vice versa). The digital code may be generated by selectively creating individual index variations (or individual gratings) with the desired spatial periods Λ1-Λn. Other illumination, readout techniques, types of gratings, geometries, materials, etc. may be used as discussed in the aforementioned patent application.
Referring to
For the images in
Referring to
The maximum number of resolvable bits N, which is equal to the number of different grating pitches Λ (and hence the number of codes), that can be accurately read (or resolved) using side-illumination and side-reading of the grating 12 in the substrate 10, is determined by numerous factors, including: the beam width w incident on the substrate (and the corresponding substrate length L and grating length Lg), the thickness or diameter D of the grating 12, the wavelength λ of incident light, the beam divergence angle θR, and the width of the Bragg envelope θB (discussed more in the aforementioned patent application), and may be determined by the equation:
Referring to
In this case, each bit (or Λ) is defined by whether its corresponding wavelength falls within the Bragg envelope, not by its angular position within the Bragg envelope 200. As a result, it is not limited by the number of angles that can fit in the Bragg envelope 200 for a given composite grating 12, as in the embodiment discussed hereinbefore. Thus, using multiple wavelengths, the only limitation in the number of bits N is the maximum number of grating pitches Λ that can be superimposed and optically distinguished in wavelength space for the output beam.
Referring to
One way to measure the bits in wavelength space is to have the input light angle θi equal to the output light angle θo, which is kept at a constant value, and to provide an input wavelength λ that satisfies the diffraction condition (Eq. 1) for each grating pitch Λ. This will maximize the optical power of the output signal for each pitch Λ detected in the grating 12.
Referring to
Alternatively, the source 300 may provide a continuous broadband wavelength input signal such as that shown as a graph 316. In that case, the reflected output beam 310 signal is provided to a narrow band scanning filter 318 through a lens 321 which scans across the desired range of wavelengths and provides a filtered output optical signal 320 to the reader 308. The filter 318 provides a sync signal on a line 322 to the reader, which is indicative of which wavelengths are being provided on the output signal 320 to the reader and may be similar to the sync signal discussed hereinbefore on the line 306 from the source 300. In this case, the source 300 does not need to provide a sync signal because the input optical signal 24 is continuous. Alternatively, instead of having the scanning filter being located in the path of the output beam 310, the scanning filter may be located in the path of the input beam 24 as indicated by the dashed box 324, which provides the sync signal on a line 323.
Alternatively, instead of the scanning filters 318,324, the reader 308 may be a known optical spectrometer (such as a known spectrum analyzer), capable of measuring the wavelength of the output light.
The desired values for the input wavelengths λ (or wavelength range) for the input signal 24 from the source 300 may be determined from the Bragg condition of Eq. 1, for a given grating spacing Λ and equal angles for the input light θi and the angle light θo. Solving Eq. 1 for λ and plugging in m=1, gives:
λ=Λ[sin(θo)+sin(θi)] Eq. 7
It is also possible to combine the angular-based code detection with the wavelength-based code detection, both discussed hereinbefore. In this case, each readout wavelength is associated with a predetermined number of bits within the Bragg envelope. Bits (or grating pitches Λ) written for different wavelengths do not show up unless the correct wavelength is used.
Accordingly, the bits (or grating pitches Λ) can be read using one wavelength and many angles, many wavelengths and one angle, or many wavelengths and many angles.
Referring to
It should be understood that there is still a trade-off discussed hereinbefore with beam divergence angle θR and the incident beam width (or length L of the substrate), but the accessible angular space is theoretically now 90 degrees. Also, for maximum efficiency, the phase shift between adjacent minimum and maximum refractive index values of the grating 12 should approach a π phase shift; however, other phase shifts may be used.
In this case, rather than having the input light 24 coming in at the conventional Bragg input angle θi, as discussed hereinbefore and indicated by a dashed line 701, the grating 12 is illuminated with the input light 24 oriented on a line 705 orthogonal to the longitudinal grating vector 704. The input beam 24 will split into two (or more) beams of equal amplitude, where the exit angle θo can be determined from Eq. 1 with the input angle θi=0 (normal to the longitudinal axis of the grating 12).
In particular, from Eq. 1, for a given grating pitch Λ1, the +/−1st order beams (m=+1 and m=−1), corresponds to output beams 700,702, respectively. The +/−2nd order beams (m=+2 and m=−2), corresponds to output beams 704,706, respectively. The 0th order (undiffracted) beam (m=0) corresponds to beam 708 and passes straight through the substrate. The output beams 700-708 project spectral spots or peaks 710-718, respectively, along a common plane, shown from the side by a line 709, which is parallel to the upper surface of the substrate 10.
For example, for a grating pitch Λ=1.0 um, and an input wavelength λ=400 nm, the exit angles θo are ˜+/−23.6 degrees (for m=+/−1), and +/−53.1 degrees (from m=+/−2), from Eq. 1. It should be understood that for certain wavelengths, certain orders (e.g., m=+/−2) may be reflected back toward the input side or otherwise not detectable at the output side of the grating 12.
Alternatively, one can use only the +/−1st order (m=+/−1) output beams for the code, in which case there would be only 2 peaks to detect, 712, 714. Alternatively, one can also use any one or more pairs from any order output beam that is capable of being detected. Alternatively, instead of using a pair of output peaks for a given order, an individual peak may be used.
Referring to
Thus, for a given pitch Λ (or bit) in a grating, a set of spectral peaks will appear at a specific location in space. Thus, each different pitch corresponds to a different elevation or output angle which corresponds to a predetermined set of spectral peaks. Accordingly, the presence or absence of a particular peak or set of spectral peaks defines the code.
In general, if the angle of the grating 12 is not properly aligned with respect to the mechanical longitudinal axis of the substrate 10, the readout angles may no longer be symmetric, leading to possible difficulties in readout. With a thin grating, the angular sensitivity to the alignment of the longitudinal axis of the substrate 10 to the input angle θi of incident radiation is reduced or eliminated. In particular, the input light can be oriented along substantially any angle θi with respect to the grating 12 without causing output signal degradation, due the large Bragg angle envelope. Also, if the incident beam 24 is normal to the substrate 10, the grating 12 can be oriented at any rotational (or azimuthal) angle without causing output signal degradation. However, in each of these cases, changing the incident angle θi will affect the output angle θo of the reflected light in a predetermined predictable way, thereby allowing for accurate output code signal detection or compensation.
Referring to
In addition, the azimuthal multiplexing can be combined with the elevation or output angle multiplexing discussed hereinbefore to provide two levels of multiplexing. Accordingly, for a thin grating, the number of bits can be multiplexed based on the number of grating pitches Λ and/or geometrically by the orientation of the grating pitches.
Furthermore, if the input light angle θi is normal to the substrate 10, the edges of the substrate 10 no longer scatter light from the incident angle into the "code angular space", as discussed herein and/or in the aforementioned patent application.
Also, in the thin grating geometry, a continuous broadband wavelength source may be used as the optical source if desired.
Referring to
Referring to
Referring to
In the case where incident light 610 is incident along the same direction as the grating vector (Kb) 207, i.e., θi=0 degrees, the incident light sees the whole length Lg of the grating 12 and the grating provides a reflected output light angle θo=0 degrees, and the Bragg envelope 612 becomes extremely narrow, as the narrowing effect discussed above reaches a limit. In that case, the relationship between a given pitch Λin the grating 12 and the wavelength of reflection λ is governed by a known "Bragg grating" relation:
λ=2neffΛ Eq. 8
where neff is the effective index of refraction of the substrate, λ is the input (and output wavelength) and Λ is the pitch. This relation, as is known, may be derived from Eq. 1 where θi=θo=90 degrees.
In that case, the code information is readable only in the spectral wavelength of the reflected beam, similar to that discussed hereinbefore for wavelength based code reading. Accordingly, the input signal in this case may be a scanned wavelength source or a broadband wavelength source. In addition, as discussed hereinbefore for wavelength based code reading, the code information may be obtained in reflection from the reflected beam 614 or in transmission by the transmitted beam 616 that passes through the grating 12.
It should be understood that for shapes of the substrate 10 or element 8 other than a cylinder, the effect of various different shapes on the propagation of input light through the element 8, substrate 10, and/or grating 12, and the associated reflection angles, can be determined using known optical physics including Snell's Law, shown below:
nin sin θin =nout sin θout Eq. 9
where nin is the refractive index of the first (input) medium, and nout is the refractive index of the second (output) medium, and θin and θout are measured from a line 620 normal to an incident surface 622.
Referring to
If an optical waveguide is used any standard waveguide may be used, e.g., a standard telecommunication single mode optical fiber (125 micron diameter or 80 micron diameter fiber with about a 8-10 micron diameter), or a larger diameter waveguide (greater than 0.5 mm diameter), such as is describe in U.S. patent application Ser. No. 09/455,868, filed Dec. 6, 1999, entitled "Large Diameter Waveguide, Grating". Further, any type of optical waveguide may be used for the optical substrate 10, such as, a multi-mode, birefringent, polarization maintaining, polarizing, multi-core, multi-cladding, or microstructured optical waveguide, or a flat or planar waveguide (where the waveguide is rectangular shaped), or other waveguides.
Referring to
Referring to
The grating 12 may be impressed in the substrate 10 by any technique for writing, impressed, embedded, imprinted, or otherwise forming a diffraction grating in the volume of or on a surface of a substrate 10. Examples of some known techniques are described in U.S. Pat. Nos. 4,725,110 and 4,807,950, entitled "Method for Impressing Gratings Within Fiber Optics", to Glenn et al; and U.S. Pat. No. 5,388,173, entitled "Method and Apparatus for Forming Aperiodic Gratings in Optical Fibers", to Glenn, respectively, and U.S. Pat. No. 5,367,588, entitled "Method of Fabricating Bragg Gratings Using a Silica Glass Phase Grating Mask and Mask Used by Same", to Hill, and U.S. Pat. Nos. 3,916,182, entitled "Periodic Dielectric Waveguide Filter", Dabby et al, and U.S. Pat. No. 3,891,302, entitled "Method of Filtering Modes in Optical Waveguides", to Dabby et al, which are all incorporated herein by reference to the extent necessary to understand the present invention.
Alternatively, instead of the grating 12 being impressed within the substrate material, the grating 12 may be partially or totally created by etching or otherwise altering the outer surface geometry of the substrate to create a corrugated or varying surface geometry of the substrate, such as is described in U.S. Pat. No. 3,891,302, entitled "Method of Filtering Modes in Optical Waveguides", to Dabby et al, which is incorporated herein by reference to the extent necessary to understand the present invention, provided the resultant optical refractive profile for the desired code is created.
Further, alternatively, the grating 12 may be made by depositing dielectric layers onto the substrate, similar to the way a known thin film filter is created, so as to create the desired resultant optical refractive profile for the desired code.
The substrate 10 (and/or the element 8) may have end-view cross-sectional shapes other than circular, such as square, rectangular, elliptical, clam-shell, D-shaped, or other shapes, and may have side-view sectional shapes other than rectangular, such as circular, square, elliptical, clam-shell, D-shaped, or other shapes. Also, 3D geometries other than a cylinder may be used, such as a sphere, a cube, a pyramid or any other 3D shape. Alternatively, the substrate 10 may have a geometry that is a combination of one or more of the foregoing shapes.
The shape of the element 8 and the size of the incident beam may be made to minimize any end scatter off the end face(s) of the element 8, as is discussed herein and/or in the aforementioned patent application. Accordingly, to minimize such scatter, the incident beam 24 may be oval shaped where the narrow portion of the oval is smaller than the diameter D1, and the long portion of the oval is smaller than the length L of the element 8. Alternatively, the shape of the end faces may be rounded or other shapes or may be coated with an antireflective coating.
It should be understood that the size of any given dimension for the region 20 of the grating 12 may be less than any corresponding dimension of the substrate 10. For example, if the grating 12 has dimensions of length Lg, depth Dg, and width Wg, and the substrate 12 has different dimensions of length L, depth D, and width W, the dimensions of the grating 12 may be less than that of the substrate 12. Thus, the grating 12, may be embedded within or part of a much larger substrate 12. Also, the element 8 may be embedded or formed in or on a larger object for identification of the object.
The dimensions, geometries, materials, and material properties of the substrate 10 are selected such that the desired optical and material properties are met for a given application. The resolution and range for the optical codes are scalable by controlling these parameters as discussed herein and/or in the aforementioned patent application.
Referring to
Also, the substrate 10 may be made of a material that is less dense than certain fluid (liquids and/or gas) solutions, thereby allowing the elements 8 to float or be buoyant or partially buoyant. Also, the substrate may be made of a porous material, such as controlled pore glass (CPG) or other porous material, which may also reduce the density of the element 8 and may make the element 8 buoyant or partially-buoyant in certain fluids.
Referring to
Referring to
Referring to
Referring to
Referring to
Referring to
Referring to
Referring to
Referring to
Referring to
Referring to
Referring to
The slide or chip may be a slide within a housing, discussed herein, or merely a slide having gooves, such as shown in
Referring to
Referring to FIGS. 38,39,40,41,52-53,54-57, regarding the grooved slide, plate or chip that the beads may be placed in.
Referring to
As shown, the microbead elements 8 are placed in the tray 200 with grooves 205 to allow the elements 8 to be aligned in a predetermined direction for illumination and reading/detection as discussed herein. Alternatively, the grooves 205 may have holes 210 that provide suction to keep the elements 8 in position. In operation, in response to incident light 212 provided perpendicular to the plane of the tray 200, the element 8 reflects light 214; while in response to incident light 216 provided oblique to the plane of the tray 200, the element 8 reflects light 218.
Regarding the formation of the grooves, the grooves in the groove plate may be made in many different ways, including being formed by SU8 photoresistant material, mechanically machining; deep reactive ion etching; or injection molding. One advantage of the injection molding approach is that the plate can be manufactured in volume at relatively low cost, and disposed of after the information about the beads is gathered in the assay process. The groove plate may be made of glass, including fused silica, low fluorescence glass, borosilicate glass. Silicon is used because it is reflective so a reflective coating is typically not needed. Alternative, a mirror coating can be applied to the plate material to achieve the desired reflectivity.
Referring to
Referring to
The groove width (w) should be at least as wide as the diameter of the bead (D) but not larger than D+15 μm.
The thickness of the depth of the groove (T) should be at least 0.5 times the diameter of the bead so that it sufficiently traps a bead once it falls into the groove even when it is subjected to mechanical agitation. The depth should not exceed 1.5 times the diameter of the bead so as to prevent more than one bead from falling into the same groove location.
Groove plates have been made using a thick photoresist called SU8 and is available from Microchem. The resist is both chemically inert and mechanically robust once fully cured. The groove walls are formed by the resist material, which is deposited onto a glass or substrate. Advantages of this process include the ability to tailor the depth of groove by controlling the thickness of the resist material, and virtually every other geometric attribute through the design of the photo mask. Because it is photolithographic process, essentially any shape profile can be made. For example grooves can be made in simple rows, concentric circles, or spirals. Other features such as discrete wells, spots and cross hatches can be made as fiducial marks for tracking and positional registration purposes.
The scope of the invention is also intended to include the grooves having a flat bottom as shown in
Referring to
Referring to
Referring to
Consistent with that discussed herein, the grooved plate 200 may be made of glass or plastic or any material that is transparent to the code reading incident beam 24 and code reading output light beams 27 as well as the fluorescent excitation beam 800 and the output fluorescent optical signal 802, and is properly suited for the desired application or experiment, e.g., temperature range, harsh chemicals, or other application specific requirements.
The code signal 822 from the bead code reader 820 and the fluorescent signal 810 from the fluorescence detector are provided to a known computer 812. The computer 812 reads the code associated with each bead and determines the chemical probe that was attached thereto from a predetermined table that correlates a predetermined relationship between the bead code and the attached probed. In addition, the computer 812 and reads the fluorescence associated with each bead and determines the sample or analyte that is attached to the bead from a predetermined table that correlates a predetermined relationship between the fluorescence tag and the analyte attached thereto. The computer 812 then determines information about the analyte and/or the probe as well as about the bonding of the analyte to the probe, and provides such information on a display, printout, storage medium or other interface to an operator, scientist or database for review and/or analysis, consistent with shown in step 4 of
Alternatively, instead of having the code excitation source 801 and the fluorescence excitation source 803, the reader 24 may have only one source beam which provides both the reflected optical signal 27 for determining the code and the fluorescence signal 802 for reading the tagged analyte attached to the beads 8. In that case the input optical signal is a common wavelength that performs both functions simultaneously, or sequentially, if desired.
The microbeads 8 may be coated with the desired probe compound, chemical, or molecule prior to being placed in the grooved plate 200. Alternatively, the beads 8 may be coated with the probe after being placed in the grooved plate 200. As discussed hereinbefore, the probe material may be an Oligo, cDNA, polymer, or any other desired probe compound, chemical, cell, or molecule for performing an assay.
The scope of the invention is not intended to be limited to using or detecting fluorescent molecule markers during the assay process. For example, embodiments of the invention are envisioned using and detection other types of molecular markers in other types of processes.
Referring to
In summary, the closed format approach provides a method for effectively distributing and aligning microbeads during the readout process, as described below:
The basic process for handling microbeads with a curvette for readout consists of the following steps:
(3) After the cuvette is inverted and the microbeads flow out onto the groove plate side of the cuvette, the microbeads naturally align in the grooves via a small amount of rocking or agitation, which forms part the assay process described herein.
One of the key advantages of using the cuvette device is that the potential to nearly index match the glass microbeads with a buffer solution thereby reducing the divergence of the laser beam caused by the lensing effect of the microbeads, and minimizing scatter form the groove plate itself.
Another advantage involves the potential to prevent microbeads from ever stacking up on top of each other, by limiting the space between the bottom and the top plate to be less than twice the diameter of the microbeads.
Another advantage is that the cover keeps the fluid from evaporating.
An alternative embodiment of the closed approach, which involves sectioning the closed region into two regions, one where the microbeads are free to move about in a plane, either in a groove or not, and a second region where the microbeads are trapped in a groove and can only move along the axes of a groove. Trapping the microbeads in a groove is accomplished by further reducing the height of the chamber to the extent that the microbeads can no longer hop out of a groove. In this embodiment, the free region is used to pre-align the microbeads into a groove, facilitating the introduction of microbeads into the trapped section. By tilting this type of cuvette up gravity can be used to pull the microbeads along a groove from the free region to the trapped region. Once in the trapped region the microbeads move to the end of the groove where they stop. Subsequent microbeads will begin to stack up until the groove is completely full of microbeads, which are stacked head to tail. This has the advantage of packing a large number of microbeads into a small area and prevents the microbeads from ever jumping out of the grooves. This approach could also be used to align the microbeads prior to injection into some form of flow cytometer, or a dispensing apparatus.
The dimensions and geometries for any of the embodiments described herein are merely for illustrative purposes and, as such, any other dimensions may be used if desired, depending on the application, size, performance, manufacturing requirements, or other factors, in view of the teachings herein.
It should be understood that, unless stated otherwise herein, any of the features, characteristics, alternatives or modifications described regarding a particular embodiment herein may also be applied, used, or incorporated with any other embodiment described herein. Also, the drawings herein are not drawn to scale.
Although the invention has been described and illustrated with respect to exemplary embodiments thereof, the foregoing and various other additions and omissions may be made therein and thereto without departing from the spirit and scope of the present invention.
|Citing Patent
|Filing date
|Publication date
|Jan 26, 2011
|Dec 18, 2012
|Illumina, Inc.
|Optical reader system for substrates having an optically readable code
|Sep 14, 2009
|Jul 30, 2013
|Illumina, Inc.
|Composition including an item and an encoded optical substrate and a method for identifying an item
|Sep 23, 2011
|Oct 22, 2013
|Illumina, Inc.
|Optical system and method for reading encoded microbeads
|Sep 7, 2010
|Dec 24, 2013
|Illumina, Inc.
|Elongated microparticles having an optically detectable code configured to at least one of reflect or filter light
|Aug 18, 2005
|Mar 16, 2006
|Illumina, Inc.
|Diffraction grating-based encoded element having a substance disposed thereon
|Jan 13, 2014
|May 8, 2014
|Snu R&Db Foundation
|Assay method using encoded particle-based platform
|U.S. Classification
|International Classification
|Cooperative Classification
|European Classification
|Jun 16, 2011
Owner name: ILLUMINA, INC., CALIFORNIA
Free format text: MERGER;ASSIGNOR:CYVERA CORPORATION;REEL/FRAME:026461/0284
Effective date: 20080509
Owner name: CYVERA CORPORATION, CONNECTICUT
Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNORS:MOON, JOHN A.;KERSEY, ALAN D.;PUTNAM, MARTIN A.;AND OTHERS;SIGNING DATES FROM 20040811 TO 20040817;REEL/FRAME:026478/0658
|Apr 15, 2015
|Fee payment
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A low profile version of the Floyd Rose Original.
The Floyd Rose Pro Series Tremolo Bridge with R2 Nut is a low profile version of the Floyd Rose Original and has narrower string spacing. The Original Floyd Rose Tremolo string spacing is .420 inches and the Floyd Rose Pro strings spacing is .400 inches. No copy can duplicate the double-locking design of the Floyd Rose Tremolo System that allows you to lock your guitar in tune at the nut and at the bridge. The Floyd Rose Tremolo System is engineered like no other tremolo system. Its high quality steel parts are manufactured to exacting specifications.
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Comments about Floyd Rose Pro Series Tremolo Bridge with R2 Nut:
The tremolo DOES NOT come with any installation instructions or templates to help you install it into your guitar. If you are good at measuring and doing intermediate skilled wood work you can do it yourself. You will need a drill press and a small router to complete the installation. I used the sawdust and gorilla glue to fill the old bridge post holes. I recommend that you have a professional guitar tech install the tremolo especially if it is going into an expensive guitar.
I love the low profile design. The improved string locking block holds more of the string. I haven't had a string pull out and I have been trying make it happen.
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