IP Library Granted Patent US 9,329,129
Granted Patent B2
US 9,329,129 · App. 14/440,864 · Granted May 3, 2016

Method, kit and system for imaging a blood sample

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Quick Facts
Patent No.
US 9,329,129
App. No.
14/440,864
Granted
May 3, 2016
Kind
B2
Abstract

Provided is a method for imaging a blood sample and a kit and system for executing the method. The method includes introducing a cell suspension including red blood cells onto a base surface of a carrier having a vertical height (H) being greater than or equal to a vertical depth (h) of the cell suspension when on the base carrier, the cell suspension including a cell concentration (C) being determined by a defined function; allowing the cells in the cell suspension to settle on the base surface of the carrier to form a monolayer of cells thereon; and acquiring at least one microscope image of at least a portion of the monolayer of cells; wherein the at least one microscope image is obtained by a microscope set to Depth Of Field that is not more than 20% of the vertical height of the cell suspension settled on the base surface.

Claims (56)

1. A method for imaging a blood sample, the method comprising:

introducing a cell suspension comprising red blood cells, onto a base surface of a carrier having a vertical height (H) being greater than or equal to a vertical depth (h) of said cell suspension when on said base carrier, the cell suspension comprising a cell concentration (C) being determined by the function:

C=F /( h *pi/4* d 2 )

(F) being a desired base surface coverage; and (d) being an average cell dimension of the cells in the cell suspension;

allowing the cells in the cell suspension to settle on said base surface of the carrier to form on the base surface of the carrier a monolayer of cells; and

acquiring at least one microscope image of at least a portion of the monolayer of cells,

wherein said at least one microscope image is obtained by a microscope set to Depth Of Field (DOF) that is not more than 20% of the vertical height of the cell suspension when settled on said base surface.

2. The method of claim 1 , wherein said vertical depth (h) is between 20μm and 1000μm.

3. The method of claim 2 , wherein said DOF is between 0.5 μm and 10μm.

4. The method of claim 1 , wherein said at least one microscope image is obtained by a microscope to a DOF that is not more than 15% of the vertical height of the cell suspension when settled on said base surface.

5. The method of claim 1 , further comprising, prior to introducing, diluting a blood sample comprising the cells by a dilution factor (D) to obtain the cell suspension, said factor D being a function of a concentration of cells (C 0 ) per one ml before dilution and said desired concentration C.

6. The method of claim 5 , wherein said factor D is determined by the function:

D

=

C

0

F

/

(

h

*

pi

/

4

*

d

2

)

7. The method of claim 5 , wherein sample is a blood sample and said factor D is between 50 and 300.

8. The method of claim 1 , comprising diluting a blood sample comprising red blood cells to obtain a cell concentration C in a cell suspension such that after setting on the base surface, the cells' density is between about 10,000 to about 30,000 cells per mm 2 .

9. The method of claim 8 , wherein the cell suspension forms a monolayer having an average base surface coverage of between 0.4 and 0.9.

10. The method of claim 1 , comprising selecting a microscope objective lens that provides said DOF, wherein said lens permits acquiring an image of at least one object being no more than 3μm long at any dimension thereof.

11. The method of claim 10 , wherein said lens permits acquiring an image of at least one object having a height with respect to said base surface of no more than 3μm.

12. The method of claim 1 , further comprising analyzing said at least one image to determine based thereon presence or absence of a pathogen in said cell suspension.

13. The method of claim 12 , wherein at least part of said cells are infected with a blood infecting protozoa.

14. The method of claim 13 , wherein said blood infecting protozoa is selected from the genus consisting of Trypanosoma, Plasmodium; Toxoplasma and Babesia.

15. The method of claim 1 , further comprising allowing said cells to settle on said base surface for a period of time is about 1 second per μm of vertical depth h whereby said monolayer is formed.

16. The method of claim 1 , comprising allowing a period of time of less than 5 minutes for said cells to settle on said base surface as a monolayer.

17. The method of claim 1 , wherein said vertical height H is between 20μm to 300μm.

18. The method of claim 1 , wherein:

said vertical depth h of said cell suspension is between 20μm and 300μm; and

said cells are human cells comprising at least 75% red blood cells and at least part of said cells are infected with plasmodium selected from the group consisting of Plasmodium falciparum ( P. falciparum ), Plasmodium vivax ( P. vivax ), Plasmodium ovale ( P. ovale ), Plasmodium malariae ( P. malariae ), and Plasmodium knowlesi ( P.knowlesi );

wherein the method comprises mixing the cells with one or more stains comprising at least one fluorescent stain either before introducing the cell suspension onto the carrier, before allowing the cells to settle, or before acquiring said at least one microscope image and wherein the at least one microscope image comprises a fluorescent image for detecting staining with said florescent stain.

19. The method of claim 1 , further comprising acquiring a plurality of microscope images, at least two of which are provided under different conditions, said conditions comprising imaging different portions of the base surface and different illumination conditions.

20. A system for imaging a blood sample, the system comprising:

one or more reservoir units for holding, respectively, one or more sample treatment agents comprising at least one blood cells diluting agent;

a blood sample preparing unit being in fluid communication with said one or more reservoir units and configured to receive a blood sample comprising red blood cells and amount of at least one blood cell diluting agent and to form therefrom a blood cells suspension, the amount of sad at least one cell diluting agent being determined so as to dilute said sample of cells by a dilution factor (D) so as to provide a cell concentration (C);

a microscope image acquisition unit for acquiring at least one image of the blood cells suspension when on a base surface of a carrier, the carrier having a vertical height (H) being greater or equal to a vertical depth (h) of said cell suspension when on said base surface;

a controller being configured to

provide dilution factor D of diluting sad sample, factor D being a function of the desired base surface coverage (F), the average cell dimension d of cell blood cells, and the vertical depth h of said suspension of cells that provides a monolayer of the cells when settled on said base surface of the carrier; and

acquire at least one microscope image of the cell suspension by a microscope set to a Depth Of Field {DOF) that is not more than 20% of the vertical height of the cells suspension when settled on said base surface.

21. The system of claim 20 , wherein said controller is configured to cause the microscope image acquisition unit to acquire a plurality of microscope images of the monolayer, at least two of which are provided under different conditions, said conditions comprising different portions of the monolayer and different illumination conditions.

Assignments (5)
RELEASE OF INTELLECTUAL PROPERTY SECURITY AGREEMENT Recorded Jan 23, 2025
From: KREOS CAPITAL VII AGGREGATOR SCSP
To: S.D. SIGHT DIAGNOSTICS LTD.
Reel/Frame 069993/0260 →
SECURITY INTEREST Recorded Dec 7, 2023
From: S.D. SIGHT DIAGNOSTICS LTD
To: KREOS CAPITAL VII AGGREGATOR SCSP
Reel/Frame 065788/0001 →
RELEASE OF SECURITY INTEREST Recorded Nov 28, 2023
From: BANK LEUMI LE-ISRAEL B.M.
To: S.D. SIGHT DIAGNOSTICS LTD.
Reel/Frame 065673/0880 →
SECURITY INTEREST Recorded Jul 3, 2022
From: S.D. SIGHT DIAGNOSTICS LTD.
To: BANK LEUMI LE-ISRAEL B.M.
Reel/Frame 060393/0580 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded May 5, 2015
From: POLLAK, JOSEPH JOEL; HOURI YAFIN, ARNON; SALPETER, SETH J.
To: S.D. SIGHT DIAGNOSTICS LTD.
Reel/Frame 035570/0203 →