DEVICE FOR PERFORMING A BLOOD, CELL, AND/OR PATHOGEN COUNT AND METHODS FOR USE THEREOF

• US 20130273524A1
• Filed: 04/12/2013
• Published: 10/17/2013
• Est. Priority Date: 04/17/2012
• Status: Active Grant

• Current Assignee: I-Calq LLC
• Original Assignee: I-Calq LLC
• Inventors: Ehrenkranz, Joel R. L.
• Granted Patent: US 9,322,767 B2
• Time in Patent Office: Days
• US Class Current: 435/5
• CPC Class Codes: B01L 3/5027 by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip B01L3/5023 takes precedence; micromixers B01F33/30; microreactors for synthesis B01J19/0093; microcapillary devices in general B81B1/00
• CPC Class Codes: G01N 15/1433 using image recognition
• CPC Class Codes: G01N 15/1484 microstructural devices
• CPC Class Codes: G01N 2015/1006 for cytology
• CPC Class Codes: G01N 2015/1486 Counting the particles
• CPC Class Codes: G01N 2035/00881 network configurations
• CPC Class Codes: G01N 2035/00891 Displaying information to the operator
• CPC Class Codes: G01N 21/17 Systems in which incident light is modified in accordance with the properties of the material investigated where the material investigated is optically excited causing a change in wavelength of the incident light G01N21/63
• CPC Class Codes: G01N 21/8483 Investigating reagent band test-element handling not specific to a test method G01N33/4875; analytical elements specific to chemical analysis of biological material G01N33/52; autometer with reagent band G01N35/04

Abstract

Devices and methods for performing a point of care blood, cell, and/or pathogen count or a similar blood test. Disclosed herein are systems that can be used to provide rapid, accurate, affordable laboratory-quality testing at the point of care. The systems described herein are capable of imaging and counting individual cells in a prepared cell sample (e.g., a peripheral blood smear or a blood sample prepared in a microfluidic device) or another prepared cell-containing sample without the need for a microscope or other expensive and cumbersome optics. The systems described herein are designed to eliminate or replace expensive, centralized clinical testing equipment and technical personnel. Such systems may include automated data reporting and decision support.

Claims

1. A system for performing an automated cell count, comprising:

a testing device having data collection and data analysis capabilities and the ability to interface with a means for performing a blood, cell, and/or pathogen count;

a testing apparatus communicatively coupled to the testing device, wherein the testing device includes an image sensor, and a port configured such that the means for performing the blood, cell, and/or pathogen count can be inserted into the testing apparatus and positioned in proximity to an image sensor;

the testing apparatus further including a light source capable of transmitting at least one wavelength of light configured to allow the image sensor to capture at least one image of the means for performing a blood, cell, and/or pathogen count, wherein the image sensor is configured to collect an image of sufficient resolution for discerning individual cells in the image taken of the means for performing the blood, cell, and/or pathogen count; and

an automated counting system stored in a computer readable format and electronically coupled to the testing device, wherein the automated counting system is configured to count individual cells in at least a portion of the image and classify them according to cell type.

2. The system of claim 1, wherein the pixel size of the image sensor is in a range from about 1 μ

m to about 10 μ

m.

3. The system of claim 1, wherein the pixel size of the image sensor is in a range from about 4 μ

m to about 8 μ

m.

4. The system of claim 1, wherein the testing apparatus includes at least one indexing feature configured to align the testing apparatus with the testing device.

5. The system of claim 1, wherein the housing includes one or more seals to create a light-tight environment between the testing device and the housing and between the housing and the slide.

6. The system of claim 1, wherein the testing device is selected from the group consisting of a digital camera device, a cellular phone, a smart phone, and a tablet computer.

7. The testing system of claim 6, wherein the testing device is capable of communicating with a cellular telephone network.

8. The system of claim 1, wherein the light source is at least one of a camera flash, an autofocus illuminator, ambient light, sunlight, an LED light, an incandescent lamp, or a gas-discharge lamp.

9. The system of claim 1, wherein at least one of a lens, a wavelength filter, a light conducting fiber, or a light guide is interposed between the light source and the image sensor.

10. The system of claim 1, wherein the system is configured for performing a test selected from the group consisting of a complete blood count, a white blood cell count, a CD4⁺ T lymphocyte test, identifying abnormal blood cells, identifying infectious disease pathogens within blood cells, identifying different classes of lymphocytes, using other body fluids (e.g. cerebrospinal fluid, urine, sputum, joint fluid, mucus) for cellular analysis, using tissue aspirates suspended in a liquid for cellular analysis, and combinations thereof.

11. The system of claim 1, wherein the means for performing a blood, cell, and/or pathogen count is one of a peripheral blood smear or a microfluidic device configured for performing a cell count.

12. The system of claim 11, wherein the microfluidic device includes a body that includes a counting chamber configured for performing a cell count and at least one fluid channel in fluid communication with the counting chamber for introducing a sample into the counting chamber, wherein the counting chamber includes a fixative for immobilizing one or more cells present in the sample in the counting chamber.

13. The system of claim 12, wherein the at least one fluid channel further comprises (1) a sample port and a dilution port in fluid communication with a dilution chamber, the dilution chamber being in fluid communication with the counting chamber, (2) a stain port in fluid communication with the counting chamber and configured for introducing at least one cell contrast stain into the counting chamber, (3) at least one wash port in fluid communication with the counting chamber and configured for introducing at least one washing buffer into the counting chamber, and (4) a waste chamber in fluid communication with the counting chamber and configured for capturing waste fluid from the counting chamber.

14. The system of claim 12, wherein the at least one fluid channel further includes (1) a first reservoir containing a predetermined amount of a dilution buffer in fluid communication with a dilution chamber, the dilution chamber being in fluid communication with the counting chamber, (2) a second reservoir containing a predetermined amount of a stain in fluid communication with the counting chamber, and (3) at least a third reservoir containing a predetermined amount of a wash buffer in fluid communication with the counting chamber.

15. The system of claim 14, wherein one or more of the first reservoir, the second reservoir, and the at least third reservoir include blister packs configured to be punctured for successively introducing each of the dilution buffer, the stain, and the wash buffer into the at least one fluid channel.

16. The system of claim 15, wherein the dilution buffer includes at least one tracer substance configured for determining the amount of dilution buffer added to the sample.

17. The system of claim 12, wherein the counting chamber includes a multitude of intersecting grid lines defining a multitude of counting zones.

18. The system of claim 12, wherein the at least one fluid channel further comprises at least one of

one or more gate valves positioned and configured to direct movement of fluid through the least one fluid channel;

a fixative for fixing cells in the counting chamber, or

a means for drying the cells.

19. The system of claim 12, wherein the at least one fluid channel further comprises a sample reservoir configured for measuring a predetermined volume of the sample.

20. The system of claim 12, wherein the at least one fluid channel further comprises a hemoglobin determination cell in fluid communication with the at least one fluid channel and separate from the counting chamber.

21. A method for performing an automated cell count, comprising:

providing a means for performing a blood, cell, and/or pathogen count;

preparing a cell sample for counting using the means;

providing a testing device having data collection and data analysis capabilities and the ability to interface with the means for means for performing the blood, cell, and/or pathogen count;

inserting the means for means for performing the blood, cell, or pathogen count into a testing apparatus that is communicatively coupled to the testing device, wherein the testing apparatus includes;

an image sensor, and

a port configured such that the means for means for performing the blood, cell, and/or pathogen count can be inserted into the testing apparatus and positioned in proximity to the image sensor;

illuminating the cell sample prepared with the means with at least one wavelength of light configured to allow the image sensor to capture at least one image of the cell sample, wherein the image sensor is configured to collect an image of sufficient resolution for discerning individual cells in the image taken of the cell sample prepared on the means; and

querying an automated counting system stored in a computer readable format and electronically coupled to the testing device, wherein the automated counting system is configured to count individual cells in at least a portion of the image of the cell sample and classify them according to cell type.

23. The method of claim 21, further comprising staining cells in the cell sample, wherein the stain includes at least one detectable label and/or at least one cell-specific antibody selected from the group consisting of a CD4 antibody, lymphocyte typing antibodies, antibodies for detecting bacterial, viral, or parasitic pathogens, antibodies for detecting leukemic cells, antibodies for detecting erythrocyte precursors, antibodies for detecting platelets, antibodies for detecting circulating tumor cells, and combinations thereof.

24. The method of claim 21, further comprising combining at least one detectable label and/or at least one cell-specific antibody with a cell sample prior to preparing the cell sample with the means.

25. The method of claim 24, wherein the at least one cell-specific antibody is selected from the group consisting of a CD4 antibody, lymphocyte typing antibodies, antibodies for detecting bacterial, viral, or parasitic pathogens, antibodies for detecting leukemic cells, antibodies for detecting erythrocyte precursors, antibodies for detecting platelets, antibodies for detecting circulating tumor cells, and combinations thereof.

26. The method of claim 24, wherein the at least one cell-specific antibody is coupled to at least one detectable label selected from the group consisting of colored beads, colloidal gold, colloidal silver, dyes, fluorescent dyes, quantum dots, and combinations thereof.

27. A system for performing an automated blood, cell, and/or pathogen count, comprising:

a microfluidic device configured for performing a blood, cell, and/or pathogen count, wherein the microfluidic device includes a counting chamber configured for performing a complete blood, cell, and/or pathogen count and at least one fluid channel in fluid communication with the counting chamber for introducing a cell sample into the counting chamber, wherein the counting chamber includes a fixative for immobilizing blood cells the counting chamber;

a testing apparatus communicatively coupled to a computing device, wherein the testing apparatus includes a port configured such that the microfluidic device can be inserted into the testing apparatus, one or more optical elements selected and positioned for magnifying an image of one or more blood cells in the counting chamber onto an image sensor, and at least one conduit allowing data transfer back and forth between the image sensor and the computing device;

a light source capable of transmitting at least one wavelength of light configured to allow the image sensor to capture at least one image of at least a portion of the counting chamber of the microfluidic device, wherein the image sensor has a pixel size sufficient for discerning individual cells in the image taken of the at least a portion of the counting chamber of the microfluidic device; and

an automated counting system stored in a computer readable format and electronically coupled to the computing device, wherein the automated counting system is configured to count individual cells in the image taken of the at least a portion of the counting chamber of the microfluidic device and classify them according to cell type.