KR20030068780A - Bio-cell chip - Google Patents
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- KR20030068780A KR20030068780A KR1020020008394A KR20020008394A KR20030068780A KR 20030068780 A KR20030068780 A KR 20030068780A KR 1020020008394 A KR1020020008394 A KR 1020020008394A KR 20020008394 A KR20020008394 A KR 20020008394A KR 20030068780 A KR20030068780 A KR 20030068780A
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Abstract
Description
본 발명은 바이오-셀 칩 및 그의 제조 방법에 관한 것이다.The present invention relates to a bio-cell chip and a method of manufacturing the same.
최근 유전자 상세 정보가 대량으로 공개되면서 유전자의 대량 검색 (screening) 방법에 대한 요구가 증가하고 있다. 널리 알려진 예로 고밀도 cDNA 어레이 (high density cDNA array)나 DNA 칩은 동시에 수천개의 유전자를 분석하는 것을 가능하게 하였다. 이렇게 강력한 방법들이 개발되면서 임상적으로 게놈 지도 발견 (genomic discovery)를 임상적 진단이나 예후 판정, 환자개인에 대한 맞춤의학, 질병소인 유전자를 가진 사람에 대한 예방적 조치 등에 이용할 수 있도록 하는 것이 가장 급선무로 대두되었다. 조직학적으로 같은 형태를 가진 암이라도 암을 유발시키거나 진행시키는데 관여된 유전자의 변화는 다른 경우가 많고, 각각의 유전자 변화마다 임상적 예후, 항암제에 대한 반응 등이 다르므로, 이를 예측하기 위해서는 환자에서 일차적으로 얻어진 암세포나 암조직을 대상으로 다량의 유전자를 분석하는 것이 필수적이다.As gene details are recently released in large quantities, there is an increasing demand for methods for mass screening of genes. In well-known examples, high density cDNA arrays or DNA chips have made it possible to analyze thousands of genes simultaneously. With the development of these powerful methods, it is of utmost importance that clinical genomic discovery be used for clinical diagnosis or prognosis, personalized medicine for patients, and preventive measures for people with disease predisposition genes. It came up with. Even in histologically identical cancers, the genes involved in inducing or progressing cancer are often different, and the clinical prognosis and response to anticancer drugs are different for each gene change. It is essential to analyze a large amount of genes in cancer cells or cancer tissues obtained primarily from.
현재 모든 생명공학연구는 세포의 형태가 파괴된 후 DNA, RNA, 단백질 등이세포에서 분리된 상태로 분석되고 있으며, 이들이 세포내에서 가지는 위치적 및 생리학적 유의성 (positional & biological significance)은 알 수 없다. 세포 자체를 그대로 유지한 상태에서 DNA, RNA, 단백질 등을 분석한다면 생물학적 의미를 가진 정보를 얻을 수 있다. 그러나 세포라는 검체는 우선 보존이 용이치 않고 여러 종류의 분석에는 많은 숫자의 세포가 필요하므로 연구에 제약이 있었다.Currently, all biotechnological studies have analyzed DNA, RNA, and proteins separated from cells after cell morphology has been destroyed, and their positional and physiological significance in cells is unknown. none. If you analyze DNA, RNA, protein, etc. while keeping the cell itself, you can get information with biological meaning. However, the sample of cells was not easy to preserve, and the study was limited because a large number of cells were required for various types of analysis.
환자의 검체로 유전자분석을 할 경우, 그 양이 풍부한 조직과는 달리, 혈액종양이나 환자의 몸에서 떨어져 나온 암세포(cytology specimen)는 그 양이 적어 여러가지 유전자 분석을 하기에 양적 제한이 많았다. 극소량의 세포로 세포정렬 (cell array)를 제작할 수 있다면 검체량 부족도 해결하고 동시에 수많은 유전자를 저가의 비용으로 분석할 수 있게 된다.When genetic analysis was performed on a patient's specimen, unlike abundant tissues, blood tumors or cancer cells that came off the patient's body were small in quantity, and therefore, there were many quantitative limitations for various genetic analysis. If cell arrays can be produced with very few cells, this can resolve the lack of sample volume and at the same time analyze many genes at low cost.
본 발명자들은, 세포를 작은 공간에 스팟 (spot) 형식으로 분사하여 정렬 (array) 형식으로 제작한다면 소량의 검체로도 수백가지의 실험을 할 수 있고, 수백개의 대상에 대해 동일한 조건으로 실험을 시행할 수 있음에 착안하여 본 발명을 완성하게 되었다.The present inventors can perform hundreds of experiments even with a small amount of samples if the cells are sprayed in a small space into a spot format and are produced in an array format. With this in mind, the present invention has been completed.
본 발명은 수천개의 세포에 대해 동시에 유전자 발현 (gene expression) 분석이 가능하도록 하는 데에 그 목적이 있다.An object of the present invention is to enable gene expression analysis on thousands of cells at the same time.
본 발명은 바이오-셀 칩 및 그의 제조방법에 관한 것이다.The present invention relates to a bio-cell chip and a method of manufacturing the same.
본 발명의 바이오-셀 칩은, 기존의 DNA 칩의 DNA 대신에 세포를 정렬하여 고정시킨 것이다. 즉 많은 수의 세포를 작은 공간에 정렬하여 고정시킨 바이오 칩이다. 본 발명에 따른 바이오-셀 칩에는 예를들면 70mmX30mm 정도의 작은공간에 100~2000개 정도, 바람직하게는 100~500개의 세포를 정렬하여 고정시킬 수 있다.Bio-cell chip of the present invention, by aligning the cells instead of the DNA of the conventional DNA chip It is fixed. That is, a biochip in which a large number of cells are aligned and fixed in a small space. In the bio-cell chip according to the present invention, for example, about 100-2000 cells, preferably 100-500 cells, can be fixed in a small space of about 70 mm × 30 mm.
본 발명의 바이오-셀 칩을 형성하는 고정물질 즉 세포를 최종적으로 정렬하여 고정될 칩판은 기존의 바이오 칩에서 사용되는 것을 그대로 사용할 수 있다. 예를들면 자연형광정도 (background fluorescence)가 낮은 유리슬라이드, 플라스틱, 실리콘 등을 열거할 수 있다.Fixing material forming the bio-cell chip of the present invention, that is, the chip plate to be finally aligned by fixing the cells can be used as it is used in existing biochips. For example, glass slides, plastics, and silicon with low background fluorescence can be listed.
본 발명의 바이오-셀 칩의 제조는, 기존의 DNA 칩 등의 제조방법에 따라 제조할 수 있다. 그러나 기존의 DNA, RNA, 단백질 등을 이용한 바이오 칩 제조와 달리 세포의 경우 고정액내에 보관된 액체 상태의 검체가 퍼져나가지 않게 하여야 작은 공간에 많은 수의 세포를 정렬하여 고정시킬 수 있다. 이를 위한 특별한 조치가 요구된다.The production of the bio-cell chip of the present invention can be produced according to a production method such as an existing DNA chip. However, unlike conventional biochip manufacturing using DNA, RNA, and protein, cells can be fixed by arranging a large number of cells in a small space only when the liquid sample stored in the fixed solution is not spread. Special measures are required for this.
이에 본 발명자는 각 세포들이 각각 독립되어 고정될 수 있도록 각각의 (room)을 형성시키는 방법으로 해결하였다. 각각의 방을 형성시키는 방법으로는The present inventors solved the method of forming each room so that each cell can be fixed independently. How to make each room
(가) 검체와 검체 사이에 장막 (septum)을 설치하거나(A) install a septum between the specimens or
(나) 칩을 형성하는 고정물질 표면, 즉 칩판의 표면을 일정한 모양으로 깎아 검체가 분주될 수 있는 웰 (well)을 만들거나(B) The surface of the fixed material forming the chip, that is, the surface of the chipboard, is shaved to form a well in which a sample can be dispensed.
(다) 칩을 형성하는 고정물질 표면 즉 칩판의 표편에 홈을 파는 방법 등이 있다. 웰과 홈의 차이는, 웰의 깊이는 원하는 만큼 깊게 만드는 것이 불가능하므로 그러한 점을 해결하고자 가로의 길이를 길게 깍은 경우를 홈이라 한다. 따라서 한 검체당 좀더 많은 양의 세포가 도포될 필요가 있을 경우에는 홈을 파는 방법을 적용하는 것이 바람직하다. 장막을 설치하기 위한 물질로는 고무 시멘트 (rubber cement)와 같은 시멘트 (cement), 스티커 (sticker) 등을 들 수 있다. 이들 물질은 바이오-셀 칩 제작 후 어려움 없이 완전히 제거될 수 있는 물질이어야하며, 동소교잡염색이나 세포의 모양에 아무런 영향도 끼치지 않는 것이라야 한다.(C) There is a method of digging grooves on the surface of the fixed material forming the chip, that is, the surface of the chipboard. The difference between the well and the groove is that it is impossible to make the depth of the well as deep as desired so that the length of the horizontal cut is long to solve such a problem. Therefore, when a larger amount of cells need to be applied per sample, it is preferable to apply the digging method. Materials for installing the curtain include cement, such as rubber cement, a sticker, and the like. These materials should be able to be completely removed without difficulty after fabrication of the bio-cell chip, and should not have any effect on the in situ hybridization staining or the shape of the cells.
상기와 같이 방을 형성시키는 방법 이외에도 칩을 형성하는 고정물질 표면 즉 칩판 표면에 액체상태의 검체가 잘 퍼지지 않도록 표면 특성을 변경시킬 수 있는 특수처리를 한 후 검체를 분사하는 방법도 가능하다. 이때 특수처리는 예를 들면 폴리머와 같이 끈적끈적한 물질로 표면처리를 하는 것 등이다.In addition to the method of forming a room as described above, it is also possible to spray the sample after a special treatment that can change the surface characteristics so that the liquid sample does not spread well on the surface of the fixed material forming the chip, that is, the surface of the chip plate. At this time, the special treatment is, for example, surface treatment with a sticky material such as a polymer.
따라서 본 발명의 바이오-셀 칩은, 세포들이Therefore, the bio-cell chip of the present invention,
(1) 각각 독립적인 방에 위치하여 고정되거나(1) each located in a separate room and fixed
(2) 칩판 표면에 처리된 끈적끈적한 물질에 의해 각각 독립적으로 정렬하여 고정되어 있을 수 있다. 이때 상기의 방은 장막, 웰 또는 칩판의 표면에 파놓은 홈에 의해 형성된 것일 수 있으며, 상기의 장막은 시멘트 또는 스티커로 이루어진 것일 수 있다.(2) The sticky materials treated on the surface of the chipboard may be aligned and fixed independently of each other. In this case, the room may be formed by grooves dug on the surface of the curtain, the well or the chipboard, and the curtain may be made of cement or a sticker.
본 발명의 바이오-셀 칩 제조에서 검체 즉 세포의 분사방식으로서는, 자동화된 로보트 시스템을 운영한다면 500개 이상 또는 2000개 정도의 검체를 동시에 로딩 (loading) 하는 것이 가능하며 다음과 같은 두가지 방식으로 운영할 수 있다. 우선 슬라이드에 500개 이상 또는 2000개 정도의 각각 다른 검체를 로딩하고 동소교잡을 위한 시약은 한 종류로 도포할 수 있다. 또 다른 방법은 각각의 다른 검체위에 각각 다른 동소교잡 반응 시약을 극소량 자동화 시스템을 이용하여 원하는 검체위에 반응시키는 방법도 있다.In the bio-cell chip manufacturing of the present invention, as a method of spraying a sample, a cell, if an automated robotic system is operated, it is possible to simultaneously load 500 or 2000 samples and operate in the following two ways. can do. First, more than 500 or 2000 different samples may be loaded onto the slide, and one kind of reagent for in situ hybridization may be applied. Another method is to react different orthogonal reaction reagents on different samples onto a desired sample by using a micro-automated system.
각각의 검체는 고유의 번호를 가지며 판독 현미경과 연결된 프로그램을 이용하여 자동으로 다시 원하는 검체를 찾아갈 수 있도록 할 수 있다. 검체의 결과 판독은 결과 정보가 입력된 프로그램을 이용하여 슬라이드 전체 검체를 자동으로 판독할 수 있다.Each sample has a unique number and can be automatically retrieved again using the program associated with the reading microscope. In the result reading of the specimen, the entire slide specimen may be automatically read using a program in which the result information is input.
동소교잡법에서 가장 문제가 되는 슬라이드 간 편차 (slide to slide variation)을 해결할 수 있다. 실제 연구나 환자 검사에 사용되는 검사는 슬라이드와 슬라이드간의 조건이 동일하다고 판정할 수 없어 결과판정에 문제가 되는 경우가 많다. 이 경우 한 슬라이드에 약 100개정도의 검체를 도포한 바이오-셀 칩을 이용하면 표준화 (standardization) 문제가 해결된다. 본 발명의 바이오-셀 칩은 수천개의 세포에 대해 동시에 유전자 발현 (gene expression)을 분석할 수 있도록 할 수 있는 방법이다.The slide to slide variation, which is the most problematic in the in situ hybridization, can be solved. Tests used in actual research or patient testing often fail to determine that the conditions between the slides are the same, which is often a problem in determining the results. In this case, the standardization problem is solved by using a bio-cell chip coated with about 100 samples on one slide. The bio-cell chip of the present invention is a method capable of analyzing gene expression on thousands of cells at the same time.
본 발명을 하기의 실시예 및 실험예로 더욱 구체적으로 설명하나 이로써 본 발명이 한정되는 것은 아니다.The present invention will be described in more detail with reference to the following Examples and Experimental Examples, but the present invention is not limited thereto.
실시예 1.Example 1.
수동으로 혹은 소량의 액체가 분주될 수 있는 멀티-디스펜서 (multi-dispensor)를 사용하여 한 슬라이드에 100개의 검체를 도포하여 염색체 분석 및 유전자 분석을 실시하여 소량의 검체와 소량의 시약으로 염색에 성공했다.100 specimens are applied to one slide by hand or using a multi-dispensor that can dispense a small amount of liquid. Chromosomal analysis and gene analysis have been performed to successfully dye with a small amount of sample and a small amount of reagent. did.
고무 시멘트 (rubber cement)를 사용하여 유리슬라이드에 100개의 장막을 만들었으며, 약 10분간 건조시킨후 각 웰 (well)에 검체를 1 ㎕씩 분주하였다. 20분후, 분주한 검체가 완전히 마르면 고무 시멘트를 완전히 제거하고 동소교잡 (in situ hybridization) 염색을 시행하였다.Rubber membrane (rubber cement) was used to make 100 membranes on the glass slide, and after drying for about 10 minutes, 1 μl of the sample was dispensed into each well. After 20 minutes, the sample was completely dried and rubber cement was completely removed and in situ hybridization staining was performed.
통상 사용하는 시약량은 슬라이드 한개당 10 ㎕인데 이 방법으로는 같은 시약량으로 100개의 검체를 검사할 수 있었다.The amount of reagent usually used is 10 μl per slide. This method allowed 100 samples to be tested with the same amount of reagent.
실험예 1.Experimental Example 1.
대량의 환자들은 대상으로 세포를 이용한 암검진을 할 경우 바이오-셀 칩을 이용할 수 있다. 폐암환자의 검진이나 추적후 검사로 객담의 세포를 관찰한다. 대개는 PAP 염색 (PAP staining)을 하여 일일이 판독하는데 형태학적으로는 암세포나 이형성 세포 (dysplastic cell)를 구별하지 못하는 경우가 있다. 이 경우 한국인 폐암세포의 특이적인 유전자변화를 검사한다면 진단의 특이성을 높일 수 있다. 폐암 환자의 검진에 각각의 환자의 객담에서 분리한 세포로 바이오-셀 칩을 제작하여 특이적인 유전자변화를 동소교잡법으로 검색하면 인력을 이용하지 않고도 폐암세포를 검색할 수 있는 대량검진이 가능하다. 이렇게 특별한 조치없이 몸에서 쉽게 얻을 수 있는 세포로는 소변에 떨어져 나오는 방광암 세포를 들 수 있다. 소변 검체에서 분리한 세포로 cell chip을 제작하여 방광암 특이 유전자 변화를 동소교잡법으로 검색한다면 환자가 병원에 오지 않고도 특별한 조작없이 방광암의 대량검진및 방광암 치료후 추적이 가능하다.A large number of patients can use bio-cell chips for cancer screening. Examine sputum cells by screening or follow-up examination for lung cancer patients. Usually PAP staining (PAP staining) to read one by one morphologically can not distinguish between cancer cells or dysplastic cells (dysplastic cells). In this case, specificity of the genetic changes in lung cancer cells in Korea can increase the specificity of the diagnosis. For the screening of lung cancer patients, if bio-cell chips are produced from cells separated from the sputum of each patient and searched for specific genetic changes by in situ hybridization, large-scale examinations for lung cancer cells can be performed without using manpower. . The cells that can be easily obtained from the body without any special measures include bladder cancer cells that fall off the urine. If a cell chip is made from cells isolated from urine specimens and the bladder cancer specific genetic changes are detected by in situ hybridization, patients can be followed up after mass screening of bladder cancer and treatment of bladder cancer without special manipulation without coming to the hospital.
하기의 문헌들은 본 발명자가 본 발명의 바이오-셀 칩을 이용하여 여러가지 검사를 실시한 실험예들을 발표한 논문들이다.The following documents are papers in which the inventors have published experimental examples of various tests using the bio-cell chip of the present invention.
① Kyoung un park Cha Ja She Hee Young Shin Hyo Seop Ahn Chong Jai Kim Byung Kyu Cho Han Ik ChoDong soon Lee. The Low Incidences of TEL/AML1 fusion and TEL Deletion in Korean hildhood Acute Leukemia by Extra-signal Fluorescence In Situ Hybridization. Cancer Genetics and CytogeneticsThe Low Incidences of TEL/AML1 fusion and TEL Deletion in Korean hildhood Acute Leukemia by Extra-signal Fluorescence In Situ Hybridization. Cancer Genetics and Cytogenetics, 2001, 126;73-77. (IF 1.8, 교신저자)① Kyoung un park Cha Ja She Hee Young Shin Hyo Seop Ahn Chong Jai Kim Byung Kyu Cho Han Ik Cho Dong soon Lee . The Low Incidences of TEL / AML1 fusion and TEL Deletion in Korean hildhood Acute Leukemia by Extra-signal Fluorescence In Situ Hybridization. Cancer Genetics and CytogeneticsThe Low Incidences of TEL / AML1 fusion and TEL Deletion in Korean hildhood Acute Leukemia by Extra-signal Fluorescence In Situ Hybridization. Cancer Genetics and Cytogenetics, 2001, 126; 73-77. (IF 1.8, corresponding author)
②Dong Soon Lee, Sunny Kim, Eul Ju Seo, Chan Jung Park, Hyun Sook Chi, Byoung Hak Yoon, Wo Ho Kim, Han Ik Cho. Predominance of trisomy 1q in myelodysplastic syndromes in Korea; Is it a ethnic difference? - 3 year-experience of multi-center study - Genetics and Cytogenetics, 2001,129;73-77. (IF 1.8, 제일저자)② Dong Soon Lee , Sunny Kim, Eul Ju Seo, Chan Jung Park, Hyun Sook Chi, Byoung Hak Yoon, Wo Ho Kim, Han Ik Cho. Predominance of trisomy 1q in myelodysplastic syndromes in Korea; Is it a ethnic difference? 3 year-experience of multi-center study-Genetics and Cytogenetics, 2001, 129; 73-77. (IF 1.8, first author)
③Dong Soon Lee, Eu Chong Kim, Byoung Hak Yoon, Eun Kyung Ko, Sun Yang Park, Woo Hoo Kim, Jong Hyun Yoon, Han Ik Cho. Can Minor bcr/abl Translocation in Acute Leukemia be Discriminated from Major bcr/abl by Extra-Signal FISH Analysis Haematologica 2001 86;991-992 (IF 2.4, 제일저자)③ Dong Soon Lee , Eu Chong Kim, Byoung Hak Yoon, Eun Kyung Ko, Sun Yang Park, Woo Hoo Kim, Jong Hyun Yoon, Han Ik Cho. Can Minor bcr / abl Translocation in Acute Leukemia be Discriminated from Major bcr / abl by Extra-Signal FISH Analysis Haematologica 2001 86; 991-992 (IF 2.4, first author)
④ Kyoung Un Park,Dong Soon Lee, Hye Seung Lee, Chong Jai Kim, and Han Ik Cho. Granulocytic Sarcoma inMLL-positive Infant Acute Myelogenous Leukemia: Fluorescence In Situ Hybridization Study of Childhood Acute Myelogenous Leukemia for DetectingMLLRearrangement. American Journal of Pathology. 2001:159;2001-2016 (IF 6.0, 교신저자)④ Kyoung Un Park, Dong Soon Lee , Hye Seung Lee, Chong Jai Kim, and Han Ik Cho. Granulocytic Sarcoma in MLL -positive Infant Acute Myelogenous Leukemia: Fluorescence In Situ Hybridization Study of Childhood Acute Myelogenous Leukemia for Detecting MLL Rearrangement. American Journal of Pathology. 2001: 159; 2001-2016 (IF 6.0, corresponding author)
본 발명의 바이오-셀 칩은, 작은 검체량과 작은 시약량으로 동시에 대량의 검체를 대상으로 유전자 발현 분석을 가능케 함으로써 암이나 유전병 등의 질병 검사 및 약효 검사 등에서 비용, 시간 및 노력면에서 혁신적인 진보를 실현시켰을 뿐만아니라 그 검사결과에 대한 신뢰도 또한 탁월하게 향상시킬 수 있게 되었다.The bio-cell chip of the present invention enables the gene expression analysis of a large amount of samples at the same time with a small amount of sample and a small amount of reagent, thereby making innovative advances in terms of cost, time, and effort in disease and drug tests such as cancer and genetic diseases. Not only has it been realized, but the reliability of the test results has also been greatly improved.
또한 본 발명의 바이오-셀 칩을 이용함으로써 다음과 같은 응용이 가능하게 되었다:In addition, the bio-cell chip of the present invention enables the following applications:
① 인체에서 noninvasive method로 세포를 쉽게 채취할 수 있는 종양의 대량검진에 이용한다. 예를 들면 폐암이나 방광암의 경우 객담이나 소변을 채취하여 수집한 세포로 암세포 특이 유전자 변화나 특이 항원을 염색하여 자동분석 system으로 분석하여 높은 정확도로 검색할 수 있다.① It is used for mass screening of tumors that can easily collect cells by noninvasive method in human body. For example, in the case of lung cancer or bladder cancer, cells collected by collecting sputum or urine can be detected with high accuracy by analyzing cancer cell specific gene changes or specific antigens and analyzing them with an automatic analysis system.
② 각종 종양세포주를 한 곳에 깔아놓은 종양세포주 어레이 칩 (array chip)을 제작할 수 있으며, 이울러 실험의 표준화 (standardization)가 가능하게 되었다.② Tumor cell line array chip with various tumor cell lines laid in one place can be manufactured, and standardization of experiments is now possible.
③ 환자의 종양에서 직접 채취하여 분리한 종양세포 칩을 제작할 수 있으며 아울러 기초 생명과학 연구자들에게도 제공할 수 있게 되었다.③ The tumor cell chip can be manufactured directly from patient's tumor and separated and can be provided to basic life science researchers.
④ 단일세포-PCR (single cell-PCR)의 표준화 (standardization)가 가능하게 되었다.④ Standardization of single cell-PCR is now possible.
⑥ 미세 잔존백혈병 세포의 검색이 가능하게 되었다.⑥ It was possible to search for fine residual leukemia cells.
⑦ 항암제 감수성 예측 시스템의 개발이 가능하게 되었다.⑦ Development of anticancer susceptibility prediction system is now possible.
⑧ 약물감수성 판단에 이용할 수 있으며 아울러 마이크로 세포 배양 칩 (micro cell culture chip) 제작이 가능하게 되었다.⑧ It can be used for the determination of drug sensitivity, and it is also possible to manufacture micro cell culture chips.
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| KR1020020008394A KR20030068780A (en) | 2002-02-18 | 2002-02-18 | Bio-cell chip |
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| JP2003568093A JP2005517411A (en) | 2002-02-18 | 2003-02-14 | Bio-cell chip |
| PCT/KR2003/000322 WO2003068982A1 (en) | 2002-02-18 | 2003-02-14 | Bio-cell chip |
| US10/920,054 US20050112623A1 (en) | 2002-02-18 | 2004-08-17 | Bio-cell chip |
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| JP4853905B2 (en) | 2006-05-30 | 2012-01-11 | 国立大学法人北海道大学 | Cell chip |
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| US5545531A (en) * | 1995-06-07 | 1996-08-13 | Affymax Technologies N.V. | Methods for making a device for concurrently processing multiple biological chip assays |
| US6103479A (en) * | 1996-05-30 | 2000-08-15 | Cellomics, Inc. | Miniaturized cell array methods and apparatus for cell-based screening |
| US6352827B1 (en) * | 1996-08-28 | 2002-03-05 | President And Fellows Of Harvard College | Detection of multiple nucleic acid sequences in a fluid sample |
| US6210910B1 (en) * | 1998-03-02 | 2001-04-03 | Trustees Of Tufts College | Optical fiber biosensor array comprising cell populations confined to microcavities |
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