Method for detecting fluorescent signals in a biological sample
Inventors
Kim, Young Min • Zhu, Yanning • Agarwal, Yash • Wang, Xiuzhong • Armstrong, Aaron • Borgerding, Robert • MacGinitie, Andrew • Seppo, Antti • Ichetovkin, Ilia • Kilpatrick, Michael • Tsipouras, Petros • Tafas, Triantafyllos P.
Assignees
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Publication Number
US-8791429-B2
Publication Date
2014-07-29
Expiration Date
Abstract
A method for automated microscopic analysis wherein the test protocol is obtained from interrogatable data affixed to each microscope slide. The method further comprises the algorithms that implement the test protocol.
Core Innovation
The disclosed invention provides an interrogatory method and an automated microscopy system for high throughput microscopic analysis of cancerous cells using an automated microscope. The method motorizedly transports one or more microscope slides to a microscopic stage, where each slide comprises a biological sample and electronically-interrogatable data pertaining to detectably labelled biological structures. The electronically-interrogatable data are read from the slides, and the data identify how the biological sample is to be analyzed, so the sample is analyzed situated on the slide and the analyzed data are read and reported to identify and locate cell/tissue origin and state in situ.
A core aspect is determining an analytical protocol from the electronically-interrogatable data and using that information to analyze multiply and distinguishably fluorescently labelled biological structures in an x-y-z space. The protocol directs imaging and scanning of serial focal slices in x-y-z space, including sequential analysis at low and high magnification and identification using adjustable focal settings recorded in the form of instructions. The system is operated in a concatenated series of steps that regulates focusing, optimizes the slide region, and provides suitable sequential image stacks for analysis.
The invention also specifies cancer-cell identification by tagging biological structures with detectably distinguishable FISH signals and fluorescent labels directed to DAPI and chromosomal aneuploidy probes, together with additional chromosome-specific probes. The probe set includes chromosome 3, chromosome 7, chromosome 9, and chromosome 17 probes, with fluorescence differentiation based on different excitation filters or a plurality of laser sources.
Claims Coverage
The provided excerpt includes five independent claims. Across these claims, the coverage centers on reading electronically-interrogatable data on slides to determine an analysis protocol, performing protocol-driven analysis using x-y-z image stacks with low-to-high magnification or sequential focal scanning, automated operation by motorized stage/slide transport and controller-driven instructions, and cancerous cell identification using specified FISH signals and probe targets under multicolor fluorescence differentiation.
Protocol-driven slide interrogatory high throughput microscopy
Transporting one or more microscope slides motorizedly to a microscopic stage, reading electronically-interrogatable data from the slides, determining from the electronically-interrogatable data information on how the biological sample is to be analyzed, and analyzing and reading and reporting analyzed data of detectably labelled biological structures wherein cell or tissue origin and state is identified and located in situ.
Automated microscopy controlled analysis with x-y-z image stacks
Reading electronically-interrogatable data from slides comprising a sample selected from detectably labelled biological structures in isolated cells and tissue, determining from the electronically-interrogatable data information on how the slide sample is to be analyzed in image stacks of serial focal slices in an x-y-z space, analyzing the sample using the information determined from the electronically-interrogatable data, and reading and reporting analyzed data to identify or locate multiply fluorescently labelled biological structures using a plurality of laser sources at high magnification across focal top to bottom edges of the x-y-z space in a concatenated series of steps for regulating focusing, optimizing the slide region and providing suitable sequential image stacks at low and high magnification.
Instruction-driven automated high throughput image processing system
Providing a motorized microscope stage and a slide holding mechanism configured to transport slides to the stage, reading electronically-interrogatable data and identifying a sample image stack located in an x-y-z space scanned at high magnification sequentially from top to bottom edge focal levels, using a controller to receive and determine from the electronically-interrogatable data information on how the biological sample is to be analyzed and providing instructions, and using an analyzing module responsive to the controller instructions to analyze electronically-interrogatable data of the biological sample, wherein cancerous cell biological structures are identified by tagging with specified FISH signals directed to DAPI, chromosomal aneuploidy probes, and chromosome-3/7/9/17 probes.
Database protocol instruction automated microscopy focusing and digital imaging
Electronically reading slide-based electronically-interrogatable data, using database protocol instructions to determine information on how multiply and distinguishably fluorescently labelled biological structures in isolated nuclei is to be analyzed, focusing one or more objective lens components on a sample or on a focal plane comprised in an x-y-z space while analyzing according to instructions from electronically-interrogatable data, and capturing a digital image of the sample or a portion of the sample while analyzing according to the instructions, wherein cancerous cell biological structures are identified by tagging with different laser excitable FISH signals directed to DAPI, chromosomal aneuploidy probes, and chromosome-3/7/9/17 probes with differentiation based on different excitation filters or laser sources.
Non-transitory computer-readable medium program for high throughput automated microscopy with iterative cassettes and slides
Providing a non-transitory computer-readable storage medium embodying a program of instructions executable by a computer for a method of high throughput microscopic analysis that uses an automated microscope with slide stage, objective lens, slide cassette, programmable means for operating the microscope according to a protocol, and programmable means for providing an analytical outcome, wherein the program includes transporting a first cassette, transporting a first slide from the cassette to the stage, interrogating data on the slide to provide information related to a protocol for analysis of detectably probe labelled biological structures, positioning the slide on a slide stage, causing the microscope to analyze the sample in accordance with the analytical protocol encoded in the interrogatable data, providing an analytical outcome representing the sample with distinguishably labelled structures located at low magnification followed by verification of type of cancerous cells at high magnification, and repeating the instructions across remaining slides and then across remaining cassettes.
Across the independent claims, the inventive coverage is concentrated in automated, protocol-driven microscopy in which electronically-interrogatable slide data are read to determine analysis instructions, those instructions drive x-y-z scanning and image stack acquisition with sequential focal and magnification analysis, and cancerous cells are identified by tagging with specified DAPI, chromosomal aneuploidy, and chromosome 3/7/9/17 FISH probes under multicolor fluorescence differentiation.
Stated Advantages
Not explicitly described in patent.
Documented Applications
Not explicitly described in patent.
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