Blood and cell analysis using an imaging flow cytometer

• US 8,660,332 B2
• Filed: 03/13/2013
• Issued: 02/25/2014
• 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: Ortyn, William E., Basiji, David A., Morrissey, Philip J., George, Thaddeus C., Hall, Brian E., Zimmerman, Cathleen A., Perry, David J.
• Primary Examiner(s): Johns, Andrew W
• Application Number:
  US13/798,442
• Publication Number:
  US 20130201317A1
• Time in Patent Office: 349 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: A61B 1/00188 with focusing or zooming features
• 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 15/1475 using image analysis for extracting features of the particle
• CPC Class Codes: G01N 2015/1488 Methods for deciding
• CPC Class Codes: G06V 20/695 Preprocessing, e.g. image segmentation

Abstract

Multimodal or multispectral images of cells comprising a population of cells are simultaneously collected. Photometric and/or morphometric image features identifiable in the images are used to identify differences between first and second populations of cells. The differences can include changes in a relative percentage of different cell types in each population, or a change in a first type of cell present in the first population of cells and the same type of cell in the second population of cells. The changes may be indicative of a disease state, indicative of a relative effectiveness of a therapy, or indicative of a health of the person from whom the cells populations were obtained.

Claims

1. An imaging system configured to acquire and analyze image data separately collected from a first population of cells and a second population of cells, where the image data for each of the first and the second population of cells include a plurality of images of individual cells that are acquired simultaneously, to enable differences between the first population and the second population of cells 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;

(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 produces a plurality of images of the individual cell, the plurality of images comprising the image data, wherein the plurality of images includes at least one type of image selected from the group consisting of;

(i) darkfield images;

(ii) brightfield images; and

(iii) fluorescent images; 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, wherein the plurality of images comprises at least one of the following two types of images:

(a) multispectral images; and

(b) multimodal images.

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

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

5. The imaging system of claim 1, wherein the processor is further configured to identify at least one morphometric image feature that differs between the first and the second population of cells.

6. The imaging system of claim 1, wherein the processor is further configured to identify at least one photometric image feature that differs between the first and second population of cells.

7. The imaging system of claim 1, wherein the processor is further configured to quantify the difference between the first and second population of cells in regard to a distribution of a molecule within the cells of each 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 the second population of cells, but not in the other of the first and the second population of cells.

9. The imaging system of claim 1, wherein the light dispersing element comprises a plurality of filters that separate the light that has passed through the collection lens into the dispersed light, the dispersed light comprising a plurality of light beams of different wavelengths or wavebands.

10. The imaging system of claim 9, wherein the imaging lens focuses each light beam of a specific wavelength on a different portion of the detector so that the plurality of images comprise different spectral images of each of the individual cells, and the spectral images are disposed in different portions of the detector.

11. The imaging system of claim 10, wherein the processor is further configured to detect one or more markers included in the individual cells, by analyzing one or more of the spectral images.

12. The imaging system of claim 1, further comprising a light source disposed to illuminate each of the individual cells to facilitate forming at least one of a bright field image, a darkfield image, or fluorescent images, for each of the individual cells.

13. An imaging system for collecting, processing and analyzing image data simultaneously collected for individual cells in each of a plurality of different populations of cells to identify differences between individual cells of the plurality of different population of cells, comprising:

(a) a light source for illuminating individual cells of the plurality of different populations while there is relative motion between the individual cells and the imaging system, light from the individual cells traveling along a collection path and through at least one collection lens, producing collected light;

(b) an optical dispersing component disposed to receive the collected light and to disperse it as a plurality of dispersed light beams having different spectral characteristics;

(c) at least one imaging lens that is disposed to receive and focus the dispersed light beams along separate paths;

(d) a detector disposed so that the dispersed beam are simultaneously focused on different portions of the detector as a plurality of images of one or more of the individual cells currently being processed by the imaging system, the detector producing output signals corresponding to each of the plurality of images; and

(e) a processor for analyzing the output signals from the detector to determine any differences between the individual cells of the plurality of populations.

14. The imaging system of claim 13, wherein the processor analyzes the output signals to detect any difference between at least one of the following types of images of the individual cells that are included in the plurality of populations:

(a) darkfield images;

(b) brightfield images; and

(c) fluorescent images.

15. The imaging system of claim 13, wherein the processor determines differences between the individual cells in the plurality of populations based on detecting a relative abundance of one or more markers in the plurality of images of the individual cells, wherein the one or more markers are identified by the processor by analyzing the output signal from the detector in response to specific spectral images of the individual cells in which the markers are present.

16. The imaging system of claim 13, wherein the processor determines the difference between the plurality of different populations in regard to a number of cell types present in each of the plurality of different populations.

17. The imaging system of claim 13, wherein the processor identifies one or more differences in morphological characteristics of the individual cells present in the plurality of different populations.

18. The imaging system of claim 13, wherein the processor further determines a disease condition of the individual cells in at least one of the plurality of different populations based upon the differences between the individual cells of the plurality of different populations detected by the processor.

19. An imaging system configured to acquire and analyze image data separately collected from a first population of cells and a second population of cells, where the image data for each of the first and the second population of cells include a plurality of images of individual cells that are acquired simultaneously, to enable differences between the first population and the second population of cells 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;

(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 produces 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, wherein the processor is further configured to determine a disease condition in one of the first population of cells and the second population of cells based upon the at least one difference between the first population of cells and the second population of cells.