Blood and cell analysis using an imaging flow cytometer

• US 7,925,069 B2
• Filed: 09/29/2009
• Issued: 04/12/2011
• Est. Priority Date: 01/25/1999
• Status: Expired due to Fees

• Current Assignee: Cytek Biosciences Inc
• Original Assignee: Amnis Corporation (DiaSorin S.p.A.)
• Inventors: Hall, Brian, George, Thaddeus, Zimmerman, Cathleen, Morrissey, Philip, Basiji, David A., Ortyn, William E., Perry, David
• Primary Examiner(s): Johns; Andrew W
• Application Number:
  US12/569,612
• Publication Number:
  US 20100021039A1
• Time in Patent Office: 560 Days
• Field of Search: 382/128, 382/133, 382/134, 382/321, 356/39, 356/326, 356/328, 356/417, 356/418, 356/419, 359/633, 359/634
• US Class Current: 382/133
• CPC Class Codes: G01J 3/2889 Rapid scan spectrometers; Time resolved spectrometry
• CPC Class Codes: G01J 3/36 Investigating two or more bands of a spectrum by separate detectors
• CPC Class Codes: G01J 3/4406 Fluorescence spectrometry
• CPC Class Codes: G01N 15/1429 Signal processing
• CPC Class Codes: G01N 15/1433 using image recognition
• CPC Class Codes: G01N 15/147 the analysis being performed on a sample stream
• CPC Class Codes: G01N 2015/1006 for cytology
• CPC Class Codes: G01N 2015/1497 Particle shape
• CPC Class Codes: G01N 2021/6421 Measuring at two or more wavelengths
• CPC Class Codes: G01N 2021/6441 with two or more labels
• CPC Class Codes: G01N 21/53 within a flowing fluid, e.g. smoke
• CPC Class Codes: G01N 21/6428 Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" in vivo A61B5/00; immunoassay G01N33/53
• CPC Class Codes: G01N 21/6456 Spatial resolved fluorescence measurements; Imaging
• CPC Class Codes: G01N 21/6458 Fluorescence microscopy fluorescence microscopes per se G02B21/0076 and G02B21/16
• CPC Class Codes: G01N 33/5005 involving human or animal cells immunoassay G01N33/56966; immunoassays of protozoa G01N33/56905; protozoa in screening assays C12Q1/025
• CPC Class Codes: G01N 33/505 involving T-cells
• CPC Class Codes: G06V 20/69 Microscopic objects, e.g. biological cells or cellular parts

Abstract

Multimodal/multispectral images of a population of cells are simultaneously collected. Photometric and/or morphometric features identifiable in the images are used to separate the population of cells into a plurality of subpopulations. Where the population of cells includes diseased cells and healthy cells, the images can be separated into a healthy subpopulation, and a diseased subpopulation. Where the population of cells does not include diseased cells, one or more ratios of different cell types in patients not having a disease condition can be compared to the corresponding ratios in patients having the disease condition, enabling the disease condition to be detected. For example, blood cells can be separated into different types based on their images, and an increase in the number of lymphocytes, a phenomenon associated with chronic lymphocytic leukemia, can readily be detected.

Claims

1. An imaging system configured to acquire and analyze image data collected from a first and second population of cells, where the image data includes a plurality of images of individual cells that are acquired simultaneously, to enable differences between the first and second populations to be quantified, comprising:

(a) a collection lens disposed so that light traveling from an individual cell passes through the collection lens and travels along a collection path;

(b) a light dispersing element disposed in the collection path so as to disperse the light that has passed through the collection lens into a plurality of light beams, thereby producing dispersed light, such that each light beam has a different spectral content than the other light beams;

(c) an imaging lens disposed to focus the dispersed light, producing focused dispersed light;

(d) a detector disposed to receive the focused dispersed light, such that the focused dispersed light incident on the detector simultaneously comprises a plurality of images of the individual cell, the plurality of images comprising the image data; and

(e) a processor configured to analyze the image data for the plurality of images collected from individual cells in the first and second population of cells, to quantify at least one difference between the first population of cells and the second population of cells.

2. The imaging system of claim 1, further comprising:

(a) a laser light source that produces ultraviolet light; and

(b) optical elements configured to facilitate diffraction-limited imaging performance in the 400-460 nm spectral band.

3. The imaging system of claim 1, wherein the plurality of images comprise both of the following two types of images:

(a) multispectral images; and

(b) multimodal images.

4. The imaging system of claim 1, wherein the processor is further configured to quantify a difference between a distribution of cell types in the first and second population of cells.

5. The imaging system of claim 1, wherein the processor is further configured to quantify a difference between a first type of cell present in the first population of cells and the same type of cell present in the second population of cells.

6. The imaging system of claim 1, wherein the processor is further configured to identify at least one morphometric parameter that varies between the first and second population of cells.

7. The imaging system of claim 1, wherein the processor is further configured to identify at least one photometric parameter that varies between the first and second population of cells.

8. The imaging system of claim 1, wherein the processor is further configured to identify at least one cell type present in one of the first and second population of cells, but not in the other of the first and second population of cells.

9. A method for detecting at least one difference between a first and second population of cells using image data, where the image data include a plurality of images of individual cells that are acquired simultaneously, comprising the steps of:

(a) imaging the first population of cells and the second population of cells to collect image data, such that a plurality of images for each cell are simultaneously collected, the plurality of images comprising multispectral images; and

(b) analyzing the image data collected to identify at least one difference between the first and second population of cells.

10. The method of claim 9, further comprising the step of determining whether the difference between the first and second population of cells is indicative of a health of a person from which the first and second population of cells were obtained.

11. The method of claim 9, wherein the step of imaging the first and second population of cells to obtain the image data comprises the step of imaging the first and second population of cells while there is relative movement between each population of cells and an imaging system used to image the population of cells.

12. The method of claim 9, wherein the step of analyzing the image data collected to identify at least one difference between the first and second population of cells comprises the step of identifying at least one morphometric parameter that varies between the first and second population of cells.

13. The method of claim 9, wherein the step of analyzing the image data collected to identify at least one difference between the first and second population of cells comprises the step of identifying at least one photometric parameter that varies between the first and second population of cells.

14. The method of claim 9, wherein the step of analyzing the image data collected to identify at least one difference between the first and second population of cells comprises the step of identifying at least one cell type present in one of the first and second population of cells, but not in the other of the first and second population of cells.

15. The method of claim 9, wherein the step of analyzing the image data collected to identify at least one difference between the first and second population of cells comprises the step of quantifying a difference between a distribution of cell types in the first and second population of cells.

16. The method of claim 9, wherein the step of analyzing the image data collected to identify at least one difference between the first and second population of cells comprises the step of quantifying a difference between a first type of cell present in the first population of cells and the same type of cell present in the second population of cells.

17. The method of claim 9, further comprising the step of adding a reagent to at least one of the first and the second population of cells before imaging that population of cells, wherein the reagent comprises at least one reagent selected from the group consisting of:

(a) a label that facilitates identification of one of more cellular features;

(b) a label that facilitates identification of a diseased cell;

(c) a label that facilitates identification of an abnormal cell; and

(d) a reactive substrate having a unique optical signal, the reactive substrate being configured to selectively bind to a free bio-molecule indicative of the disease condition, such that detection of the unique optical signal in the image data indicates that the disease condition is present.

18. The method of claim 9, wherein the step of imaging the first and second population of cells to obtain the image data comprises the step of illuminating each population of cells using a light source that produces ultraviolet light.

19. The method of claim 9, wherein the step of imaging the first and second population of cells to collect the image data comprises the step of simultaneously collecting at least two types of images selected from a group consisting of the following types of images:

a brightfield image, a darkfield image, and a fluorescent image.