[특허]Method and apparatus for detection of rare cells

Method and apparatus for detection of rare cells 원문보기

IPC분류정보
국가/구분	United States(US) Patent 등록
국제특허분류(IPC7판)	C12P-001/00
출원번호	US-0498533 (2009-07-07)
등록번호	US-8450082 (2013-05-28)
발명자 / 주소	Weller, III, Albert E.
출원인 / 주소	Battelle Memorial Institute
대리인 / 주소	Klein, Richard M.
인용정보	피인용 횟수 : 0 인용 특허 : 59

초록 ▼

Fluorescent dye-tagged rare cells are disposed in a biological fluid layer contained in an annular gap (12) between an inside test tube wall (14) and a float wall (16). One or more analysis images are acquired at one or more focus depths using a microscope system (10, 10′, 10″, 10′″). Each acquired analysis image is image processed each to identify candidate rare cell images. The test tube (72) is selectively rotated. The test tube and the microscope system are selectively relatively translated along a test tube axis (75). The acquiring and image processing are repeated for a plurality of fields of view accessed by the selective rotating and selective relative translating and substantially spanning the annular gap.

대표청구항 ▼

1. A method for identifying fluorescent dye-tagged cells in a biological fluid layer using a microscope system having a field of view at a selectable focus depth, the microscope field of view being substantially smaller than an area of the biological fluid layer, the method comprising: (a) illuminating the biological fluid layer through an objective of the microscope system;(b) identifying the focus depth of the objective of the microscope system at which the illuminating produces a highest background fluorescence observed through the objective;(c) identifying a depth of the biological fluid layer based on said highest background fluorescence of the biological fluid layer caused by the fluorescent dye-tagging;(d) acquiring one or more analysis images using the microscope system at one or more focus depths at or close to the identified depth of the biological fluid layer;(e) image processing the acquired one or more analysis images to identify candidate features that may correspond to dye-tagged cells;(f) relatively moving the field of view and the biological fluid layer; and(g) repeating the identifying, acquiring, image processing, and relative moving steps to identify candidate features over the area of the biological fluid layer. 2. The method as set forth in claim 1, wherein the identifying (b) comprises: acquiring a plurality of depth-sampling images at different focus depths spanning a pre-selected encompassing depth range; andidentifying the focus depth of the objective of the microscope system at which the illuminating produces said highest background fluorescence as corresponding to the focus depth of the brightest acquired depth-sampling image. 3. The method as set forth in claim 2, wherein the one or more analysis images include at least the brightest acquired depth-sampling image. 4. The method as set forth in claim 2, wherein the one or more analysis images include at least the brightest depth-sampling image and two additional depth-sampling images having focus depths closest to the optical focus depth of the brightest depth-sampling image. 5. The method as set forth in claim 2, wherein the one or more analysis images are acquired at a higher resolution than the depth-sampling images and are acquired after the acquiring of the plurality of depth-sampling images. 6. The method as set forth in claim 2, wherein the number of depth-sampling images is greater than or equal to the pre-selected encompassing depth range divided by a depth-of-view of the microscope system. 7. The method as set forth in claim 1, wherein a thickness of the biological fluid layer is greater than a depth-of-view of the microscope system, and the acquiring of the one or more analysis images comprises: acquiring a number of analysis images greater than or equal to the thickness of the biological fluid sample divided by the depth-of-view of the fluorescent imager. 8. The method as set forth in claim 1, wherein the image processing of each analysis image comprises: matched-filtering the analysis image using a filter kernel sized to substantially match images of fluorescent dye-tagged cells in the analysis images. 9. The method as set forth in claim 8, wherein the image processing of each analysis image further comprises: thresholding the matched-filtered analysis image using a threshold computed from a signal-to-noise ratio of the analysis image to generate a corresponding binary image in which the candidate features are identified. 10. The method as set forth in claim 1, further comprising prior to step (a): disposing a biological fluid sample containing the fluorescent dye-tagged cells in a test tube;disposing a float in the test tube, the float defining an annular gap between an outer surface of the float and an inner surface of the test tube;centrifuging the test tube with the biological fluid sample and the float disposed therein, the centrifuging causing a component of the biological fluid sample including the fluorescent dye-tagged cells to segregate at the annular gap to define the biological fluid layer. 11. The method as set forth in claim 10, wherein the float includes at least two annular ridges engaging with the inner surface of the test tube to seal the annular gap, the annular ridges disengaging from the inner surface of the test tube during centrifuging to admit the segregating component of, the biological fluid sample into the annular gap. 12. The method as set forth in claim 10, wherein the biological fluid sample is a blood sample, and the segregated biological fluid layer is a buffy coat containing fluorescent dye-tagged epithelial cells.

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A	생활필수품
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Method and apparatus for detection of rare cells 원문보기

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Method and apparatus for detection of rare cells 원문보기

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