text:
U.S. Open
Looking for your favorite
Thomas Hardy Ale from 1968-2008
The label reads "In 'The Trumpet Major' Hardy wrote of Dorchester strong beer
'It was of the most beautiful color that the eye of an artist in beer could
desire; full in body, yet brisk as a volcano; piquant, yet without a twang;
luminous as an autumn sunset; free from streakiness of taste, but, finally,
rather heady".
The refurbishment of the Trumpet Major pub in Dorchester was the catalyst for
the fledgling Thomas Hardy's Ale. 1968 was the 40th anniversary of the authors'
death and what better way to commemorate it than by attempting to bring fiction
to life and creating the brew that Hardy imagined.
This beer was to be no ordinary ale. Matured in oak sherry casks for nine months
and corked in decorative pint and half pint bottles. The strength was a whopping
12% and it was bottle conditioned. Thus the legend began and bottles were laid
down with the expectation of improvement as the beer matured.
Most of the production did not consist of the larger corked bottles with their
beautiful velvet neck ribbons but small nip size crown corked bottles with a
black and gold neck medallion. This depicted a silhouette of Thomas Hardy on one
side and the brewery address on the reverse. This was to become a distinctive
feature of Hardy's Ale bottles, but it was only the 1968 edition that displayed
the date on the medallions.
Hardy's Ale was not produced again until 1974 but with the exception of 1976
appeared annually after that until the final vintage of 1999. Word of mouth
spread the tale amongst connoisseurs about this wonderful strong ale, the one
that extolled us not to drink it for 5 or 10 years and proclaimed that it would
keep for 25 years!
All production during the 1970's was of nip size bottles. Note that vintages
were not released in 1969, 1970, 1971 ,1972 , 1973 or 1976. The labels were
essentially the same as the 1968 vintages although the contents were now
imperial/metric - 180ml/6.34fl oz. Two bottlings were brewed in 1979 dated 1st
January and 1st September. Things started to change at the dawn of the 1980's.
Most of the bottles released in 1980 to 1982 were in a larger 25ml. size aimed
at an increasing export market.
In 1983 came the 1st special edition. The brew was inaugurated on 15th September
1982 to mark the completion of the Centenary development of Dorchester Brewery.
The beer was made available on 7th November 1983 exactly 150 years after Charles
Eldridge held his inaugural dinner in Dorchester. To mark the visit of the Duke
of Kent to the brewery all production was made available as Royal Hardy Ale in
33cl bottles. The bottles were numbered but without the usual year prefix.
When production of nips re-commenced in 1984 there was a new label in place. The
old parchment style was replaced by a more functional style with an image of
Thomas Hardy at the top of the label. 1987 saw the brewing of 150th Anniversary
Thomas Hardy's Ale. Sarah Eldridge had commenced brewing at the Green Dragon
brewery in 1837. The commemorative brew was matured in oak, rolled daily, hand
bottled with a full long cork. The bottles sported a red velvet sash and "A"
prefix just like the original 1968 pints and half-pints. Only 2700 bottles were
issued so surviving examples are rare. 1980 prefix "K", 1981 prefix "K", 1982
prefix "L", 1983 no prefix, 1984 prefix "M", 1985 prefix "M", 1986 prefix "N",
1987 prefix "P" (prefix "A" for 150th Anniversary), 1988 prefix "Q", 1989 prefix
"R", 1990 prefix "R"
During the second half of the 1980's the lettering and numbering became very
messy. The U.S. was importing a large share of the production and a different
prefix letter was used on nips for export. A larger 355 ml size was also
produced and these were numbered without a prefix letter.
As the 1990's were ushered in changes were again made to the bottles, presumably
as cost cutting measures. The famous neck medallion was scrapped during 1991 and
a new label was introduced with the Thomas Hardy image on the neck foil instead.
Both the old and new style labels exist for 1991 vintages. From here on the
prefix letter and number were omitted except for the final special edition
bottling of 1993. This was the Silver Anniversary brew commemorating 25 years
since the first Thomas Hardy's Ale. The 33cl bottles was now the dominant size
as production was increasingly exported to the U.S. The final nips were produced
in 1994. Sadly, Thomas Hardy's Ale was not to reach the millennium as the final
Vintage was released in 1999.
Thomas Hardy Ale(2003-2008)
The OHanlons brewery, which had "re-created" the Thomas Hardy Ale over the
past six vintages of 2003-2008, but decided to quit brewing this classic ale.
The brewer is citing too-great expense due to the slowness, pain and expense of
making this relatively rare beer which many beer lovers have called the best in
the world. The OHanlons operation is small and it is too impractical for them
to continue making Thomas Hardy, so they wish to focus on their main business.
Starting a Craft
Brewery - Brewers Library
America's Best LLC
America's Best
Odd Inns
Welcome Fisherman
Electric and Hybrid Cars
Panacea USA - Alternative Energy
Ranger - Guide to US National Parks
Top 10
Ales & Lagers
in the USA
Golden Ale
Blonde Ale
English Summer Ale
English Pale Ale
English IPA
Imperial IPA
Imperial Red Ale
English Mild Ale
English Brown
American Stout
Oatmeal Stout
Imperial Stout
Old Ale
Barley Wine
Scottish Ale
Strong Scottish
Irish Red Ale
American Amber
American Pale Ale
American IPA
American Brown Ale
German Alt
German Kolsch
German Wheat
Belgian Saison
Belgian Ale
Belgian Abbey Ale
Belgian Lambic
Belgian Witbier
American Light
American Pilsner
Premium Lager
Amber Lagers
Bohemian Pilsner
German Pilsner
Munchner Helles
Munchner Dunkel
Vienna Lager
American Cream Ale
American Wheat
American specialty wheat
Fruit/Vegetable beer
Pumpkin beer
Herb and Spice beer
Chocolate Beer
Coffee beer
Rye/Roggen Beer
Wood / Barrel
Aged beer
Aged strong beer
Wood/Barrel Stout
Smoked/Rauch beer
Specialty Ales/Lagers


text: Adding up the advantages of transfer programs There are many reasons to encourage students to transfer from a community college to a four-year institution Starting their college career at a community college and transferring to a four-year institution to obtain a more advanced degree can be one of the smartest moves high school graduates can make. A community college serves as an ideal launch pad for continuing their education, while helping students save money and maximize their college investment. How to prepare Preparation for a two- or four-year school is virtually the same. "In addition to meeting with their guidance counselor on a regular basis, students should take college prep courses and keep up with math and science," says Christine Casalinuovo-Adams, interim co-director of admissions at Monroe Community College (MCC) in Rochester, N.Y. When thinking about a community college, it's also important for high school students to begin visiting schools the summer after their junior year and apply, as well as look for scholarship opportunities, in the fall of their senior year. In January of their senior year, students and their families apply for financial aid packages. "Community college staff and faculty are very engaged with students and provide personalized attention to help them decide if a two-year school with a transfer program is the right fit for them," adds Casalinuovo-Adams. Easing the transition Transfer programs are an ideal option for just about every student for various reasons. For example, a student: • May need to become better academically prepared for a four-year institution. • May not be emotionally ready to leave home. • May wish to explore his or her options before deciding on a major. • Or, a student's family may not be financially equipped to send their child to a four-year school. "For any student whose goal is a bachelor's degree, a community college is a great place to start," explains Casalinuovo-Adams. "At MCC, we work diligently to help students decide on a major early on and to ensure all classes they take are transferable. When students maintain the required GPA, the transfer to a four-year school is seamless." 2+2 = success! MCC has a formalized 2+2 Program that allows students to easily transfer to any one of approximately 30 four-year colleges across the country. Students may also choose to attend other four-year colleges and universities. The program is best suited for first-time, full-time matriculated students who know where they wish to pursue their bachelor's degree and in what major. As part of the 2+2 Program, MCC offers a number of majors including engineering, education, social work, criminal justice, computer science, communications, biotechnology and hospitality. "Historically, community colleges have an outstanding track record of helping students prepare for transferring on to the school of their dreams," insists Casalinuovo-Adams Similar transfer programs exist in the community colleges throughout the nation, so encourage your students to find out more about the transfer programs at your local community college.They need to make sure their credits will transfer from a 2-year to a 4-year college effectively so they can graduate on time. Lou Ann Benigni-Lynch, owner of CopyLAB, is a freelance writer based in Victor, N.Y.Related articles: How to fit in at a four-year school Classes that transfer

subset: commoncrawl


text:
Desotorow Gallery is currently accepting submissions for Science of Art, a juried exhibition that explores the relationship between Art and Science. Submissions are due by August 6, 2010 at 5 pm EST. Exhibition runs August 20 - 31, 2010. Open to works of all media. Open to national and international artists (professional, emerging, and student). Download the prospectus at www.desotorow.org. For more information, email email@example.com.
Desotorow Gallery is a nonprofit organization located in Savannah, GA.


text:
Friday, Dec. 4, 2009 | 3 a.m.
Employers are often turning to social networking sites to research job candidates, a study by Challenger, Gray & Christmas, an outplacement company, found last month.
This is a chance for workers looking for a job to use those sites to their advantage.
Any tool to help laid-off Las Vegans is welcome, being as the jobless rate was 13 percent in October. Workers are having to get creative when looking for job leads.
The classified ad section in Sunday's paper showed more ads were for house and car sales than were ads placed by employers looking for help.
This means expanding your search to your network, and one of the easiest ways to do that is by letting people on Facebook, LinkedIn and Twitter know you're looking for work.
"It is important to remember that all of these technologies simply enhance the job search," Challenger CEO John Challenger said in a statement. "(Technology) will never replace the face-to-face connections that are critical to a successful search."
But Challenger advises job seekers to, at the very least, set up LinkedIn accounts and consider setting up professionally geared profiles on sites such as Facebook and Twitter. Try establishing relationships with managers who could be looking for employees.
Then, instead of run-of-the-mill postings about family vacations or what you made for dinner, focus your posts on the industry you would like to work in.
Once the domain of young people, social networking sites are gaining popularity among adults. The median age of LinkedIn users is 39 years old; for Facebook, it's 33; and for Twitter, the median age is 31, according to a recent study by Pew Internet and American Life Project.
Social networking tips from Challenger:
• Build your network: Use everyone in your personal and professional networks;
• Build your brand: Focus on the industry you'd like to work in, such as posting news items and trends;
• Advertise your job loss: Letting people know you are looking for work could open up opportunities for you, especially on Twitter where employers can follow you with ease, allowing you to cast a wider net;
• Get recommendations: On LinkedIn, ask former colleagues and bosses to post a recommendation about you. Return the favor in kind;
• Join groups: LinkedIn and Facebook allow users to join professional groups, giving you a chance to communicate with others in your field;
• Think before you tweet: Use social networking tools to market yourself. Don't post anything you may regret later.
Challenger also suggested several groups on LinkedIn that job seekers should join including the Talent Buzz, Executive Suite, Linked:HR, JobsDirectUS and Project: Get Hired.
A word of caution from Challenger:
"Perhaps the most dangerous aspect of the Internet is the permanency and pervasiveness of any and all information that finds its way there. Comments on a friend's blog, reviews on consumer sites and inside jokes made for a private audience on a social networking site's public group page are all available at the click of a mouse to potential employers."
Desperate times
Las Vegas-based Adopt-a-Rescue Pet sent out a desperate e-mail to the media Thanksgiving week.
Elizabeth Rubin, founder of the nonprofit organization, said that several of the group's donors have stopped their donations to Adopt-a-Rescue because of the recession.
The group is in the process of building an animal sanctuary and retirement home for dogs in Amargosa Valley, and had enough money promised to it from its donors until they reneged on their commitment, Rubin said.
Those plans have been put on hold, as has the adoption of dogs sitting in kill shelters. The group is also receiving dogs it had already adopted out from people who have decided to abandon them.
As part of Rubin's plea for help, she announced that area business K-9 Barracks & Bath is sponsoring a holiday kibble drive to buy food for the 300-plus dogs being held by Adopt-a-Rescue. The group goes through 270 large bags of feed every month.
A $25 donation will buy a 40-pound bag of dog food.
Even a $1 donation will help, if everyone who reads this pitches in, she said.
To donate, visit adoptarescuepet.org or call 883-0035.


text:
Court to Rule Today on Appel Graft Case
The Tel Aviv Magistrate's Court is to hand down its decision today on the bribery charges against businessman David Appel.
Appel stands accused of bribing Lod mayor Benny Regev, Giv'at Shmuel local council head Zamir Ben-Ari and a senior official in the Israel Lands Administration, Oded Tal.
Also named in the indictment, issued in 2003, is Appel's real estate firm, Migdal Hazohar Construction and Benny Tabin, who is charged with being the middleman for delivering the bribe through his firm, Eichut Hayim Beganei Aviv.
Regev has also been indicted for taking a bribe.
The first charge states that Appel and his company bribed Regev to allow him to build a mall, more residential towers, a country club and a light rail system during construction of the new Ganei Aviv neighborhood in Lod.
Appel and his company allegedly also worked to have some 1,400 dunams (350 acres) of agricultural land brought within Lod's city limits and have the land rezoned for construction.
Regev allegedly worked to promote the interests of Appel and his company and therefore they contributed NIS 1.4 million to Regev's reelection campaign, disguising the payments with fictitious receipts.
According to the second charge, Appel and Migdal Hazohar gave NIS 95 million to Givat Shmuel local council head Zamir Ben-Ari's campaign so he would promote their interests in his town and rezone land for the construction of 2,000 housing units.
The third charge states that Appel bribed Tal, acting head of the ILA's Tel Aviv district and led him to believe his connections would facilitate Tal's promotion to ILA central district chief. In return, Tal allegedly advanced Appel's and Migdal Hazohar's interests in Givat Shmuel and interened with regard to a debt Appel and his wife had for the lease on their home.
Tal was convicted in 2008 of taking a bribe from Appel in a separate trial.
Want to enjoy 'Zen' reading - with no ads and just the article? Subscribe todaySubscribe now


text:
American Savings Club
Complaint 129111 Details
- Date Occurred: 05/22/2014
- Reported Damages: $400.00
- Username: G. R. Lehman
- Email: firstname.lastname@example.com
- Location: Prescott, AZ
- !
The complaint is against an online dating profile
The complaint is a listing fraud posted on public forums or sites against an anonymous entity
The complaint is mobile text spam or smishing related against an anonymous entity
The company or person contact no longer exists
International boundaries
They called said they wanted the last four digits of bank account to verify ID.
Told them I was not interested in the club, did not want it.
Next bank statement they had taken out $400.00. This is a scam.
- Anonymous SBID #5aa14f8c77
- Posted 06/22/2014
Do You Have a Complaint?Submit a Complaint ►
There are no comments posted. Be the first to comment on this complaint
Do You Have a Complaint?
Add a complaint and spread the word. High volume complaints have a better chance to get their money back. Submit a report and join forces with others like you!Submit a Complaint Now
Business Profile Summary
Company Statistics
- Complaint Against American Savings Club
- Complaints Filed: 5
- Reported Damages: $2,192.82
What is SBID?
SBID is a unique id code that identifies the user's computer and location. SBID is used to prevent fraudulent postings and help our community find users who create duplicate user accounts.


text:
A Russian field commander Tuesday offered a $1 million bounty on the head of the Islamic separatist guerrilla commander Shamil Basayev, but that may be simply posturing. "Correspondents manage to interview Basayev without much trouble, and he�s not exactly hiding out," says Meier. "It would require a stretch of the imagination to believe that the Russian special forces don�t know where he is." Even more bizarre, perhaps, is the mounting speculation that President Boris Yeltsin is unhappy with the spectacular rise in Prime Minister Vladimir Putin�s popularity prompted by the Chechnya operation. "Even though the Kremlin�s game plan was to use the war to get Putin elected president next year, there�s now talk that Yeltsin is unhappy about his prime minister getting all the glory. That�s even raised the fear that Putin may be fired." But while Yeltsin may be quite capable of sacking his handpicked heir, he�ll probably let Putin sack Grozny first.
Battle for Chechnya Brings Out the Bizarre
The second battle for Grozny has begun. Russian tanks and infantry fought their way into the suburbs of the Chechen capital Tuesday in the face of fierce resistance by its guerrilla defenders. In 1994, Russia suffered heavy casualties and killed thousands of Chechens, both fighters and civilians in a vain attempt to take control of the capital. The latest battle may be as grim. "Some Russian leaders seem to believe that anyone who hasn�t fled Chechnya by now is fair game," says TIME Moscow correspondent Andrew Meier. "A former prime minister said Sunday that eliminating bandits and terrorists in Chechnya may require wiping out half of the population."


text:
Home > Sunglasses > Polaroid > PLD 4032/S (LWT/94)
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These sunglasses complement your look with an accessory that is a cut above the rest. True to the Edel-Optics motto "SEE AND BE SEEN", you are on a par with celebrities, impressing in every environment. The PLD 4032/S is available in the Edel-Optics online shop in other styles from Polaroid collections 2016 and 2017.
Expressive lines add character to the classical approach, making the glasses a must-have for women. Full rim frames completely enclose the lenses within the frame. For a convinced eyeglass wearer, any other frame style is ruled out, with very few exceptions. The cat-eye frame lifts the face optically and its outer tips make the frame shape particularly favourable for square faces. Plastic frames like these combine durability and are comfortable to wear. The PLD 4032/S is a very snug fit on the ears and nose. Brown is currently a trend colour and also an absolute timeless classic. The colour provides a nice contrast ratio between the skin and the colour of the eyeglass frame. Thus, the transition from eyeglasses to the face is harmonious and unobtrusive. The fact that the colour grey matches everything, also applies to sunglass lenses. They suit everything in your wardrobe and never go out of fashion. Functionally, you are, of course, on the safe side, too. With 100% UV eye protection, the sun can immediately start shining. Whether driving or out on the slopes, your own safety sometimes depends on your angle of vision. The polarizing lenses in this model block the glare from reflecting surfaces, such as water, glass or snow and best sight is guaranteed.
If you press the button "order with prescription lenses', our digital dispensing optician will install the lenses in the original tint or as you wish. Your dioptre values are the only thing we require from you.
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text:
Traditionally, debuggers have been very per thread centric. For example, a debugger most typically will assign a thread to be the current thread and any thread related debug operations you perform will be done on the selected thread. In order to work with a different thread, you need to first switch the thread context. One of the ways that you were able to switch the active thread in Visual Studio was to bring up the Threads Window and double click on the thread of interest. While this works fine for applications with relatively few threads, it becomes cumbersome for heavily multithreaded applications. Luckily, Visual Studio 2012 RC has a feature known as Parallel Watch Window that aids greatly in these scenarios.
At a high level, a watch window allows you to watch the results of expressions. The expressions can be as simple as the value of a local variable but can also be much more complicated. Imagine we have the following simple code that multiple threads execute concurrently:
private static void WorkThreadFunction()
for (int i = 0; i < max_list_len; i++)
// Do work
Nothing too fancy – simply multiple threads iterating through a list of some sort. Now, lets say that 25 threads were currently executing the function above and we want to see at which iteration ('i') each of the threads is currently executing. Previously we would have to switch the active thread context 25 times and record the value of 'i' somewhere. Instead, with Visual Studio 2012 RC we can bring up the Parallel Watch Window as shown below:
Choosing Parallel Watch Window 1 (there are four to choose from) displays the Watch Window:
By default, the Parallel Watch Window brings up all the thread currently executing in the process. In order to add new watches, we need to click "<Add Watch>" column which allows us to enter an expression. In our case, the expression is as simple as showing the local variable 'i':
As soon as the watch is added we can now see the expression evaluated across all the different threads in the watch window:
Another interesting aspect of the Parallel Watch Window is that you can sort the columns as well as adding a filter to the watch (Filter by Boolean Expression).
This is a pretty powerful and time saving feature if you spend lots of time doing multithreaded debugging.
Until next time, happy debugging!


text:
Germany is the largest economy in Europe. The country has a strong tradition of excellence within technological development, and offers a wide variety of opportunities for students wishing to study abroad. Germany more that 2 million students are enrolled out of which 2, 50,000 are international students.
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text:
|Publication number
|Publication type
|Application number
|Publication date
|28 Nov 2006
|Filing date
|9 Nov 1994
|Priority date
|12 Nov 1993
|Fee status
|Also published as
|Publication number
|Michael Catt, Paul Henry Mundill, Michael Evans Prior
|Original Assignee
|Inverness Medical Switzerland Gmbh
|Export Citation
|BiBTeX, EndNote, RefMan
|Patent Citations (112), Non-Patent Citations (72), Referenced by (33), Classifications (22), Legal Events (7)
|External Links: USPTO, USPTO Assignment, Espacenet
This invention relates to devices for reading the results of assays, to assay devices for use in conjunction with reading devices, and to methods using such devices.
An objective of the invention is to provide assay result reading devices and associated sample testing devices which can provide accurate quantitative assay information in a simple, quick and cost effective manner. Such devices can be used in a wide range of situations such as hospitals, clinics, doctors' offices, and the home. Depending on the circumstances, the analyte under investigation can also vary widely. Examples are infectious disease organisms or markers, metabolites in body fluids indicative of a change in the health or condition of a patient, and administrable or ingestable substances such as medicaments or drugs of abuse.
The invention is particularly, although not exclusively, concerned with assays which can be performed by comparatively untrained people and especially in the home.
Home-use assay devices such as pregnancy tests are now well established. In the case of a pregnancy test, which merely needs to provide the user with a "yes/no" result, the technology now available enables the assay result to be read easily by eye without the need for any ancillary equipment.
Home-use assays are intended primarily to detect physiological changes in the human body, with the objective of promoting the health, general well-being or lifestyle of the individual. The consumer is becoming increasingly health conscious, and the ability of the consumer to monitor his or her bodily functions is being encouraged. In some instances this can facilitate the interaction between the individual consumer and the medical profession (GP).
There are many assays indicative of physiological changes in the human body which currently can only be performed using sophisticated laboratory techniques. In order to provide useful information concerning the individual under test, such assays generally need to yield a result in precise numerical terms, eg. the concentration of a specific analyte in a body fluid.
Accordingly there is a need for an assay system, especially applicable to the testing of body fluid samples in the home, which combines convenience of sample testing together with simple and cost-effective numerical determination of the assay result.
Many assay devices are described in the technical literature with suggestions that the assay result can be read using optical equipment. The use of fluorescence emission, or light reflectance, is often suggested. Such techniques are mostly appropriate for use in sophisticated laboratories. In EP-A2-212599, which describes multizone analytical elements having a detectable signal concentrating zone, the suggestion is made that a detectable signal indicative of an assay result in the zone can be measured by electromagnetic radiation, such as light, transmitted through the zone. EP-A2-212599 indicates that the element can be made from porous fibrous materials, such as paper and nitrocellulose. However, no practical details are provided to indicate how an accurate measurement might be made using transmitted light.
By the invention, we have found that quantitative information can be derived by transmission reading of an assay strip or the like if the incident electromagnetic radiation is uniform across a region of the test strip which encompasses and extends beyond the test zone.
In one embodiment, the invention provides a method of "reading" the result of an assay effected by concentrating a detectable material in a comparatively small zone of a carrier in the form of a strip, sheet or layer through the thickness of which electromagnetic radiation, such as light, is transmissible, wherein at least a portion of one face of said carrier is exposed to incident electromagnetic radiation which is substantially uniform across the entire portion, said portion including said zone, and electromagnetic radiation emerging from the opposite face of said carrier is measured to determine said assay result.
Preferably, the incident electromagnetic radiation is of substantially uniform intensity.
This uniformity can be achieved, for example, by providing a columated source of electromagnetic radiation, using conventional focussing means such as lenses and light guides to provide parallel incident electromagnetic radiation which falls essentially normally across the entire exposed portion of the carrier.
However, in a more preferred embodiment of the invention, the incident electromagnetic radiation is diffuse and bathes the exposed portion of the carrier uniformly in a randomly scattered manner.
In another embodiment, the invention provides an assay device comprising a porous liquid-permeable carrier strip or sheet through the thickness of which electromagnetic radiation is transmissible diffusely, said carrier being within a casing, said carrier including at least one detection zone in which an assay result is revealed by specific binding of a detectable material directly or indirectly to a binding agent immobilised in said detection zone, detection of said material being effected as a response to said electromagnetic radiation, and said casing having electromagnetic radiation transmitting regions enabling electromagnetic energy from an external source to be passed through said device, said detection zone lying in the electromagnetic radiation path between said electromagnetic radiation transmitting regions.
Preferably, the porous carrier strip or sheet comprises paper, nitrocellulose or the like, preferably of a thickness not exceeding 1 mm.
In yet another embodiment, the invention provides an assay device and assay result reader combination, wherein:
Preferably, said receiving means incorporates interlocking means engagable with corresponding interlocking means on said device to ensure that upon receipt of said device by said reader said detection zone(s) is located and maintained in a predetermined spacial relationship relative to said reading means.
Preferably, said receiving means includes actuating means triggered by said receipt of said device, said actuating means causing said reading of said detection zone(s) to be initiated.
If the assay device is provided with a casing, it is advantageous if said device casing includes internal registration means which engages with corresponding registration means associated with said carrier such that said detection zone within said device casing is located in a predetermined spacial relationship relative to said registration means on said device casing. Preferably, said internal registration means comprises a pin or the like, engagable with a hole, indentation of the like in said carrier, said detection zone being at a predetermined location on said carrier relative to said hole or indentation.
During manufacture of said assay device, said corresponding registration means may be used to facilitate or control accurate formation, e.g. by means of reagent printing techniques, of said detection zone on said carrier. In addition, or alternatively, accurate placement of said carrier within said device casing can be facilitated or controlled by said registration.
In a further embodiment, the invention provides an assay result reader, for use in conjunction with an assay device comprising a porous liquid-permeable carrier strip or sheet through the thickness of which electromagnetic radiation is transmissible, said carrier including a detection zone in which an assay result is revealed by specific binding of a detectable material directly or indirectly to a binding agent immobilised in said detection zone, detection of said material being effected as a response to said electromagnetic radiation, said assay result reader comprising:
The assay device/reader combination can be supplied to the consumer as a single test kit. In general however, whereas the reader will be a relatively permanent unit which the consumer can use time and again (and which may be provided with an electronic memory/data-processing facility which enables the results of many sequential assays to be evaluated) the testing devices will be intended for use only once and thereafter will be discarded. Accordingly, the test devices may be supplied to the consumer separately from the reader, e.g. in multi-packs.
By ensuring precise interlocking between the testing device and the reader, and also ensuring precise registration of the location of the detection zone within the testing device itself, the testing zone will be presented to the reader in a constant pre-determined position every time a testing device is inserted into the reader. The construction of the optical system within the reader (light source and sensors) can therefore be kept as simple as possible, because it is not essential for the sensors to include any scanning facility, for example, which would otherwise be required if the exact location of the detection zone was not known. By avoiding the need for a sophisticated optical reading system, the cost of the reader/monitor may be reduced. Simplification of the optical reading system may also enable the reader/monitor to be of small size which will assist convenient and unobtrusive use in the home. Of course, a scanning facility can be included in the reader if desired.
An additional benefit of providing an internal registration system which ensures precise location of the detection zone within the test device, is that automated manufacture and quality control of the testing devices can be facilitated. Because it is envisaged, for example, in the case of an ovulation cycle monitor, that the consumer will need to use several testing devices each month, the testing devices may need to be manufactured in large numbers at low cost. Internal registration can facilitate automated manufacture and high throughput.
In principle, any electromagnetic radiation can be used to effect the transmission measurement in the invention. The electromagnetic radiation should preferably be capable of being rendered diffuse. Preferably the electromagnetic radiation is light in the visible or near-visible range. This includes infra-red light and ultra-violet light. It is generally envisaged that the detectable material used as a label in the assay is one which will interact with light in the visible or near visible range, eg. by absorption. The wavelength of the electromagnetic radiation chosen is preferably at or near a wavelength which is strongly influenced, eg. absorbed, by the label. For example, if the label is a substance which is strongly coloured, ie. visible to the naked human eye when the material is concentrated, the ideal electromagnetic radiation is light of a complementary wavelength. Particulate direct labels, for example, metallic (eg. gold) sols, non-metallic elemental (e.g. Selenium, carbon) sols, dye sols and coloured latex (polystyrene) particles are ideal examples. For instance, in the case of blue-dyed latex particles, the ideal electromagnetic radiaation is visible red light which will be strongly absorbed by the blue particles.
In a preferred embodiment of the invention, the transmitted electromagnetic radiation reaching the sensor(s) should be diffuse. The diffuseness may arise as a consequence of transmission of the electromagnetic radiation through the carrier strip or sheet, but more preferably is contributed by the source of the electromagnetic radiation emitting the energy in a highly diffuse form. In a preferred embodiment of the invention the source produces highly diffuse radiation and the carrier strip or sheet through which this radiation subsequently is transmitted is in comparative terms a much weaker diffuser.
A primary advantage of the use of diffuse light or other radiation in the context of the invention is that the reading of the assay result is much less likely to be adversely influenced by blemishes or contaminating material on the assay device. For example, dirt or scratches on the assay device in the region through which the radiation must be transmitted could strongly interfere with the accuracy of the determined result if focussed rather than diffuse light is used. By the use of a diffuse light source in accordance with the invention, it is possible to provide an assay result reader which can accurately interpret the result of an assay conducted even in an essentially transparent assay device without the assay result being adversely affected by minor contamination or damage (eg. superficial scratches) to the assay device.
In a preferred embodiment of the invention, the electromagnetic radiation from the source is pulsed. By synchronising the detectors (sensors) so that they function only in phase with the pulsed radiation source, it is possible to eliminate any background interference that might be caused by external radiation, e.g. ambient light. It is envisaged that the assays will mostly be conducted under circumstances of natural daylight or, even more often, artificial light. Artificial light is usually of a pulsed nature (typically 50–100 Hz) caused by the alternating nature of electricity supplies. By adopting a pulsed radiation source for the illumination of the assay device within the reader, the intrusion of natural daylight can be ignored. By selecting the pulse frequency such that it is sufficiently different from the prevailing artificial light, any interference due to artificial light can also be avoided. Preferably the pulse frequency of the energy should be at least about 1 kHz. An ideal pulse frequency is about 16 kHz. The electronics necessary to achieve synchronous pulsed sensing are familiar to those skilled in the art.
The use of pulsed light is very advantageous because it renders it unnecessary for the monitor to be "light tight". Not merely does this simplify the construction of the monitor but the reading of the assay result can be performed while the monitor is "open", thus simplifying the operation for the user.
The source of light or other electromagnetic radiation can comprise entirely conventional components. Ideal examples are commercially available LED's, preferably chosen to give a suitable wavelength of light that is strongly absorbed by the detectable material concentrated in the test zone(s). Light from the LED's should be passed through a strong diffuser before reaching the assay device. If desired, an array of LED's which are energised in turn can be used.
Suitable diffusers can be made, for example, from plastics materials, and are available commercially. If necessary, the light-scattering properties of the diffusing material can be enhanced by including particulate materials such as Titanium dioxide and Barium sulphate. An ideal diffusing material comprises polyester or polycarbonate, containing Titanium dioxide. A good inclusion level for the particulate material is at least about 1% by weight, preferably about 2%. By the use of a diffuser, all relevant regions of an assay strip may be measured simultaneously, and differences in light output from the source are eliminated.
The sensor(s) to detect emergent light can be conventional components such as photodiodes, e.g. silicon photodiodes.
Preferably, a second diffuser, which can be made from the same material as the primary diffuser, is located in front of the sensor(s). This ensures that the view seen by the sensor is not affected by the presence or absence of a test strip in the reading head. In consequence, the monitor can be calibrated in the absence of a test strip, and then measure an assay result in the presence of an assay strip.
By employing a uniform light source in accordance with the invention, it is possible to provide a reading system for test strips and the like which is relatively tolerant to variation in the placement of the test zone(s) from one strip to another, in the absence of a scanning sensor. Further benefits are obtained if test zone placement is controlled, as described herein.
For the purposes of enhancing the likelihood of conception, assay devices have already been marketed which enable the user to monitor the urinary concentration of lutenizing hormone (LH) which peaks sharply approximately one day in advance of ovulation. Daily testing of urinary LH concentration is conducted, for example using "dipstick" technology with the assay result being provided by a coloured end point, the intensity of the colour being proportional to LH concentration. By providing the consumer with a colour chart which enables the daily result to be compared against a standard, the "LH surge" can be detected simply by eye. Unfortunately, the monitoring of LH concentration is a very rare example of an assay relying on semi-quantitative data which is ameniable to such simple technology, being possible only because in relative concentration terms the LH surge is such a dramatic event. For most other potentially useful assays the analyte concentration changes in body fluids are much more subtle and only detectable accurately by instrumental means.
A need therefore exists to extend the currently available qualitative home-use testing technology into the area of precise quantitative testing. A convenient example, which is a logical extension of the present consumer interest in home-use pregnancy testing and ovulation prediction testing, is the extension into accurate monitoring of the ovulation cycle, not merely to enhance the likelihood of conception but indeed to provide reliable information for the purposes of contraception. Proposals have been made to analyse body fluids with this objective in mind. A common theme is to monitor periodic fluctuations in various hormone metabolite levels in urine.
The invention can be used in the determination of any body fluid analyte, especially in the monitoring of the human ovulation cycle by the determination of one or more hormones or metabolites thereof in body fluid, such as urine, for example either LH and/or estrone-3-glucuronide (E3G).
Within the preferred context of the present invention it is envisaged that a home-use sample liquid testing device will include a porous carrier material, such as a strip, through which applied sample liquid such as urine can permeate and wherein the assay result occurs by means of specific binding of a detectable material in a precisely-defined region (detection zone) of the carrier, such as a narrow line or small dot, containing an immobilized specific binding reagent. The invention is therefore concerned with ways in which localisation of a detectable material in such a detection zone can be determined accurately in a simple and cost-effective manner. Home-use devices for the analysis of urine, for example in pregnancy tests and ovulation prediction tests, are now widely available commercially. Many such devices are based on the principles of immunochromatography, and typically comprise a hollow casing constructed of plastics material containing a porous assay strip carrying pre-dosed reagents. The reagents within the device may include one or more reagents labelled with a direct label, such as a dye sol, a metallic (e.g. gold) sol, or a coloured latex (e.g. polystyrene) microparticle, which are visible to the eye when concentrated in a comparatively small test area of the strip. The user merely needs to apply a urine sample to one part of the casing to initiate the assay. The assay result becomes visible by eye within a few minutes without further action by the user. Examples of such devices aare described in EP-A-291194 and EP-A-383619, the disclosures of which are incorporated herein by reference. Sample collection is conveniently achieved by means of a bibulous member which forms part of the device and which can readily take up sample liquid, e.g. from a urine stream. Optionally the bibulous member can protrude from the casing of the device to facilitate sample application.
Other embodiments of the invention, which will be apparent from the following detailed description, include assay devices for use as part of the reader/assay device combination, methods of manufacturing such assay devices, and methods of use of such assay devices and readers.
By way of example only, assay devices and readers in accordance with the invention will now be described with reference to the accompanying drawings, of which:
Referring to
Parallel lines (102–107) of assay reagents are placed on sheet 100 prior to sub-division. For the purposes of example only, the reagents are assumed to be a first immobilised antibody in lines 102 and 107, and a second different immobilised antibody in lines 103 and 106. Reagent deposition can be by means of a "pen" 108 or the like operated on a computer-controlled "x-y" plotting mechanism (not shown) and fed with appropriate buffered reagent solution via a metered flexible tube 109. If the material of sheet 100 is nitrocellulose, reagents such as antibodies and antigens can be immobilised by simple direct application onto the nitrocellulose, followed by blocking of the sheet material, for example with albumen or polyvinyl alcohol. Following reagent deposition and blocking, two lines 104 and 105 of mobile labelled reagent, such as antigen (e.g. E3G) or another antibody (e.g. anti-LH) labelled for example with a particulate direct label such as coloured latex, can be deposited. This deposition can be for example by means of another pen (not shown). Alternatively, the labelled reagent(s) can be held in a separate porous pad or the like, rather than being applied directly to the test strip material.
In order to achieve precise location of the reagent-containing lines, each longitudinal periphery 110, 111 of sheet 100 is pierced with a plurality of identical small holes 112 each one being situated within the width of a designated strip 113. Holes 112 are made in sheet 100 prior to the deposition of any reagents. The untreated sheet is located on a frame (not shown) or similar operating surface by means of a bar 114 pressed downwardly onto each lateral periphery of the sheet. Only one of these bars is (partially) shown. Each bar has a plurality of downwardly projecting pins 115, each of which locates precisely into one of the holes 112. The tracking of the reagent-depositing pen 108 is registered precisely with the position of the bars holding the sheet, and accordingly the reagent deposition is made in a predetermined precise line relative to the perforations in the sheet.
Following all necessary reagent depositions and other treatments of the sheet, the sheet is subdivided by cutting means (not shown) into individual identical strips 101. Each individual strip therefore contains one locating hole 112 with two reagent-containing lines or reaction zones (e.g. 102 and 103) located relative to hole 112 in precise predetermined positions extending across the width of each strip. At a location remote from hole 112 is a region (e.g. 104) of the strip bearing the mobile labelled reagent. The exact position of the labelled reagent relative to the hole is not necessarily as critical as the location of the reaction zones.
By way of example only, the individual strips will typically have a length of about 40 mm to about 80 mm, and a width of about 5 mm to about 10 mm, as is already conventional in the assay device art. A reagent-containing detection zone such as the reaction zones 102 and 103, will typically be a line of width about 1 mm running laterally across the strip. A small dot, e.g. circular with diameter of about 1 mm to about 3 mm, is an alternative. The detection zone is therefore only a relatively small proportion of the total area of the strip. If appropriate for the purposes of the assay, multiple detection zones containing the same or different reagents can be placed on each strip. This may necessitate more than one labelled component being used; multiple mobile labelled components can be placed upstream on the strip or elsewhere within the device (eg. in a sample application pad or wick, as described below).
Referring to
Lower half 201 of the casing also includes a light-transmitting window or aperture 209 which, in the assembled device, lies directly opposite to the result window 205 in the upper half of the casing. Lower half of the casing also contains a depression 210 which can accommodate the bottom end of the pin or peg 208 when the two halves of the casing are placed together to make an enclosure.
In the assembled device, the act of enclosing the strip and bibulous member between the upper and lower halves of the casing causes the overlapping portions 204 and 211 of the strip and bibulous member to be crimped together to provide a good moisture-conductive junction.
It is generally envisaged that the material of the casing will be opaque, e.g. white or coloured plastics material, but the casing can be translucent or indeed transparent if desired.
Referring to
The casing of the monitor is slotted to receive the portion of the assay device incorporating the result windows. On opposing sides of the slot is a light source 303 and a reading head 304.
The slot incorporates a button or projection 305 which can fit into the depression 207 on the external face of the casing of the assay device. Precise positive location of the casing within the slot is therefore achieved. Because the depression is in a fixed position relative to the internal pin or peg 208 within the assay device, and hence the registration hole 112 in the assay strip 101, the two detection zones 102 and 103 on the strip are located in a precise position relative to the reading head. The hole in the assay strip therefore acts as a positive reference throughout the manufacture of the assay device and ensures that after the device has been used and presented to the monitor the detection zones on the strip will be in the same position relative to the reading head each time. Accordingly there is no need for the reading head to incorporate a scanning facility to locate the detection zones in each presented device.
The light source or illuminator 303 incorporates a plurality of LEDs 306 to generate light, and this shines onto the assay strip via a diffuser 307 and the observation window 209 in the lower half of the assay device casing. The light passes through the thin nitrocellulose strip 101 and exits the assay device through the result window 205 in the upper half of the casing. Immediately outside window 205 is a second diffuser 308. After passing through the second diffuser 308, the light encounters a plate 309 having a plurality of apertures 310–314. There are five apertures in total, two of which (311, 313) are adjacent to the detection zones and the others (310, 312 and 314) lie in positions on either side of these detection zone apertures. The apertures are of slit form corresponding to the detection lines on the strip. The width of each of the two apertures 311 and 313 corresponding to the detection zones themselves is double the width of each of the three other apertures, which act as controls.
The light passing through these apertures travels down a corresponding slot 315–319 in a baffle plate 320. At the far end of each slot is a light detector 321. The detectors 321 are of identical size and specification. At the front face 322 of the baffle plate 320, each slot is of the same size as the corresponding aperture. At the rear face of the baffle adjacent to the light detectors each slot is of the same size as the face of the light detector adjacent to it. Accordingly, the two slots (316, 318) associated with the detection zone apertures are parallel-sided. The three slots (315, 317 and 319) associated with the control apertures increase in size as they progress towards the light detector.
The slot in the monitor can also accommodate gripping or biasing means such as one or more spring-loaded plates or pins (not shown) to further enhance the positive location of the assay device within the slot.
Ideally, the same optical signal is derived from each aperture irrespective of the precise line position opposite the apertures. The apertures can be of different sizes to promote this objective. The dimensions of the reference zone should be chosen to correspond as closely as possible with the actual area of the detection zone on the strip.
To reduce the possibility of cross-talk between the apertures, the assay strip should be held as close as possible to the apertures when the assay device is located in the slot in the monitor.
As described above, there are five optical measurement channels in the reading device. In addition, there can be a sixth electronic reference channel that provides calibration of the electronic gains in the detector circuitry.
A typical test strip may exhibit a gradient of detectable label concentration along its length, against which the detectable label at a reaction zone must be measured. To accommodate this measurements are ideally made either side of the reaction zone on the test strip. The signal from the reaction zone can be expressed as a ratio of the total signal recorded from the two adjacent reference areas on the strip.
The five measurement channels are divided into two reaction zones and three reference zones. One reference zone, located between the two reaction zones provides a reference optical measurement to both reaction zone measurements.
A reflectance measuring system must all be mounted on one side of the test strip. To achieve the same level of compactness for a five channel reading device would require the use of (relatively) expensive custom components. A transmission design can be made entirely from commercially-available, high volume optoelectronic components, facilitating the production of a monitor that is compact and relatively cheap.
The five detectors 321 are mounted on the back face of a baffle plate. Each detector views the test strip through an aperture in the baffle. The baffle prevents light viewed through one aperture from falling on adjacent detectors, and also provides accommodation for line placement tolerance. The position of the test zone within the field of view of a detector may vary from one edge of the aperture to the other in the x-axis. Any variation in the signal arising from this effect is a function of the angular displacement relative to the centre of the measuring detector. The depth of the baffle can be chosen to control the possible angular displacement of the test zone with respect to the detector, and to maintain the accuracy of the reading.
The projection 305 is maintained in precise location with respect to the apertures. The reference pin locates into depression 207 in the test device casing. This depression is also precisely located with respect to the internal pin 208 moulded into the test device, on which the test strip is located by it's own locating hole punched through the strip. The reaction zones are precisely located with respect to the locating hole. In this manner, within manufacturing tolerances, the reaction zones are held in precise positions with respect to the apertures through which the detectors view the test strip.
The iluminator can consist of a series of LEDs embedded in or placed behind a diffusing medium which provides a uniform and diffuse illumination of the test strip covering the reference and signal zones.
The incorporation of a diffuser between the apertures and the test strip is beneficial for calibration purposes. In order to calibrate each of the optical channels in the absence of the test strip it is highly desirable that each detector is collecting light from the same areas of the illuminator as is the case when a test device is present. The diffuser can be selected to be the dominant diffuser in the optical path so that the introduction of the test strip does not contribute signifiantly to changes in the illumination distribution observed by the detectors. In addition, the diffuser element can enable the optical assembly to incorporate a 'wipe clean' surface, desirable for long-term repeated performance of the optical assembly. By modulating the intensity of the illuminator, the optical channels can be calibrated, without the aid of moveable parts, 'invisibly' to the user prior to the insertion of a test device.
The test strip can consist of an optically diffuse layer of nitrocellulose or the like, preferably sandwiched between two layers of optically clear film, e.g. of polyester such as "Mylar". The clear film protects the nitrocelluose within which the assay reactions take place. Making reflectance measurements through thin transparent films is particularly difficult because of problems arising from specular reflections. Transmission measurement allows the optics to be constructed orthogonal to the measuring suface and minimises the adverse effects of reflection.
The invention is particularly applicable to the reading of test strips made of nitrocellulose and similar diffuse membranes that preferably do not exceed about 1 mm thickness.
Turning to
Turning to
At the left hand end of the circuit board is mounted the reading head 441 which comprises a central receiving slot 442 to accommodate one end of an assay device (not shown). On the front of receiving slot 442 is an illuminator 443 and immediately opposite at the rear of the slot is an optical sensing system 444 so that light can be passed across the slot (and through a testing device when inserted) and evaluated by the sensor.
Turning to
The constituent parts of the casing can be moulded from high impact or similar plastics materials such as polystyrene and polycarbonate and held together by "push fit" clips or threaded screws or any other appropriate mechanism.
Turning to the enlarged illustration of the reading head, as seen in
Also on the rear wall 505 of the slot is a fixed locating pin 506 which must engage with a corresponding hole in an inserted assay device. Also on the rear wall 505 is a light-transmitting panel 507 which covers the optical sensors. Panel 507 extends outwardly beyond the plane of rear wall 505 of the slot and has sloping edges 508, 509 to give it a distinctive profile. At opposite ends of the front wall 510 of the slot are two pins (not seen in
These same features are illustrated in
It will be appreciated that the overall layout and general shape of the monitor can be subject to very considerable variation from that described above without departing from the scope of the invention. The general shape and layout of the reading head is dictated by the need to cooperate effectively with the assay device but this shape can be varied considerably. The layout and nature of the user accessible controls and information display features can likewise be subject to considerable variation and are dictated to a large extent by aesthetic considerations.
The detailed electronics of a monitoring device capable of assimilating, remembering and handling analyte concentration data, as well as providing the preferred electronic features of the device discussed herein, and where appropriate predicting future events, such as the fertility status in an ovulation cycle on the basis of such data, can readily be provided by those skilled in the electronics art once they have been advised of the factors that such a device must take into consideration, and the information that the device must provide for the user. By way of example only, the basic functions that may be required in such a device are outlined in
As depicted in
A timer, such as a clock 806 may be required to regulate measurements within a cycle. The microprocessor 804 processes, memorizes and interprets results in the light of previous events, particularly recorded from previous cycles. The user interface 807 will generally comprise at least means, such as a push button, which the user can operate at the commencement of a cycle to initiate the operation of the device as a whole. The power supply 808 should include means, such as a memory back-up capacitator 809, to prevent loss of historical data when it becomes necessary to replace batteries.
Information can be conveyed to the user by means of a liquid crystal or LED display, for example. If desired, information on the state of fertility can be conveyed by a simple visual indication, eg a combination of colours showing, for example, green for infertile and red for fertile. Especially if the device is intended primarily as an aid to contraception, it should "fail safe" by showing a "fertile" signal.
As described above, features 803 and 806 together correspond to feature 435 (
Transmission spectrophotometry is a widely used technique for the quantification of dye concentrations in clear liquid solutions. Commercially available spectrophotometers generally require substantial modification to make measurements on diffuse (scattering) solutions. Transmission spectrophotometry is not generally thought of an appropriate method of measuring highly diffuse samples so it is generally only adopted where an alternative approach cannot be applied. For the purposes of the invention, transmission measurement offers positive benefits over the more usual reflectance approach previously employed on test strips.
Some conventional strip assays employ reflectance measurement to assess dye concentration on the strip surface (e.g. glucose monitors). The chemistry of these assays occurs in a very thin layer on the surface of a test strip. In contrast, the chemistry of the preferred strip devices of the invention takes place throughout the thickness of the test strip. Because of variations in flow and reagent deposition, the concentration of detectable label captured at a reaction zone may differ according to depth.
Curvature, surface materials, finish and solvent effects may vary the ratio of specular to diffuse reflection. For reflectance measurements it is the diffusely reflected light from the surface of the strip that carries the signal information (i.e. that light will have interacted with the detectable label), whilst the specularly reflected light will contain no information (as this light is the component that has just bounced off the surface without interacting with the detectable label in the diffuse strip). Without resorting to relatively bulky and expensive systems, it is difficult to design a reflectance measurement system that minimises specular reflection to the extent possible with transmission measurement, especially using diffuse light as in accordance with the invention.
Reflectance systems require the use of a test surface that must be removed from the optical path for the purposes of calibration. This reference surface must not deteriorate if it is to form a part of the optical assembly. In addition, mechanical movement is required to displace such a reference material when an assay strip needs to be measured. Such problems are avoided by the invention.
In addition to the specific examples of detectable materials already mentioned herein, the invention can use as labels materials which block or reflect the electromagnetic radiation, rather than absorb it, e.g. "white" particles such as latex particles in their natural uncoloured state. Alternatively, the label can be a reactant or catalyst which participates in the generation of a radiation absorbing or radiation-blocking material, e.g. an enzyme which reacts with a substrate to produce a detectable material, such as a coloured material, in the detection zone.
The purpose of this example is to confirm that the transmission reading system of the invention enables consistent data to be obtained from a testing device.
A dual-analyte testing device, selected at random from a batch of identical devices constructed as hereinbefore described with reference to
The intensities of the two test lines respectively represented the concentrations of LH and E3G in a urine sample applied to the testing device.
The testing device was inserted and removed from the monitor 10 times. The percentage light transmission for each reading was as follows:
cv %
These results indicate that the reading system of the invention produces consistent data which is not affected significantly by any variability of test line placement when the test device is inserted in the monitor.
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|U.S. Classification
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|International Classification
|Cooperative Classification
|European Classification
|9 Jan 1995
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