HIGH THROUGHPUT MULTICHANNEL READER AND USES THEREOF

• US 20120220022A1
• Filed: 09/01/2010
• Published: 08/30/2012
• Est. Priority Date: 09/01/2009
• Status: Active Grant

• Current Assignee: Trustees of Boston University
• Original Assignee: Trustees of Boston University
• Inventors: Ehrlich, Daniel J., McKenna, Brian
• Granted Patent: US 8,936,762 B2
• Time in Patent Office: Days
• US Class Current: 435/286.2
• CPC Class Codes: G01N 15/14 Optical investigation techniques, e.g. flow cytometry
• CPC Class Codes: Y10T 436/11 Automated chemical analysis
• CPC Class Codes: Y10T 436/25 including sample preparation
• CPC Class Codes: Y10T 436/2575 Volumetric liquid transfer

Abstract

The present invention relates generally to the field of high content screening of particles, e.g., cells in a flow cytometric system. In particular, the present invention relates to devices, methods and systems to obtain line-scan images of particles, e.g., cells, of a plurality of different samples simultaneously, where the line-scan images can be used to identify cells based on at least one of a variety of phenotypic characteristics such as shape, asymmetry, and intracellular information for cell sorting and selection. In some embodiments, the line-scan images are obtained as the particles, e.g., cells, in a plurality of different samples flow through a plurality of microchannels, reducing the need and time for focusing of the image detection system. In some embodiments, the laser spot size has a small spatial resolution for rapid capturing of images of cells. In some embodiments, the laser spot size has a larger spatial resolution for imaging of larger particles or cells, e.g., rare cells in a sample. In some embodiments, the devices, methods and systems are automated for a high-throughput, high content screening of particles such as cells in a plurality of samples.

Claims

1. A line-scan imaging device for rapid image analysis of particles in multiple samples of interest, comprising:

a. a microfluidic device comprising a plurality of microchannels, wherein each microchannel has an inlet and at least one outlet;

b. a scanning detector configured to record emitted fluorescence in a portion of each of the plurality of microfluidic channels of the microfluidic device; and

c. an output device configured to receive data input from the scanning detector.

2. The line-scan imaging device of claim 1, wherein the output device comprises a storage device and/or a data analysis system that permits cell phenotype determination from the emitted fluorescence detected in step (b).

4. The line-scan imaging device of claim 1, wherein the scanning detector is selected from the group consisting of:

a 1-D image detector, a sparse 2-D image detector, a laser-induced florescence (LIF) detector, an avalanche photodiode, or a plurality of photomultipliers (PMTs) and/or photodiode detectors.

9. The line-scan imaging device of claim 1, wherein each microchannel of the plurality of microchannels is at least 30 μ

m wide in diameter.

12. The line-scan imaging device of claim 1, wherein the scanning detector records emits fluorescence at 6 μ

m intervals or less across the diameter of each microchannel of the plurality of microchannels.

14. The line-scan imaging device of claim 1, wherein the detector has (a) a detection diameter of between 1-5 μ

m parallel to the microchannel, (b) a detection diameter of less than one microchannel diameter parallel to the microchannel, or (c) a detection diameter of about 3.5 μ

m parallel to the microchannel.

17. The line-scan imaging device of claim 1, wherein the inlet of each of the plurality of microchannels fluidly connects to a plurality of sample wells for sample input into each microchannel.

19. The line-scan imaging device of claim 1, wherein the each microchannel has at least one positive outlet and at least one negative outlet to permit sorting of cells between positive and negative states.

22. The line-scan imaging device of claim 17, wherein the sample well receives samples from the output of a plurality of microfluidic channels from a parallel microfluidic cytometer.

24. The line-scan imaging device of claim 1, wherein each microchannel of the plurality of microchannels on the microfluidic device is configured for vertical hydrodynamic focusing.

39. The line-scan image device of claim 1, wherein the reference point indicating a cell phenotype is a reference line scan image of cell phenotype.

40. The line-scan image device of claim 39, wherein the reference point indicating a cell phenotype is a value from an algorithm indicating a cell phenotype.

53. The line-scan imaging device of claim 1, wherein the device comprises at least 16 microchannels.

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