Automated microscopic cell analysis
This disclosure describes single-use test cartridges, cell analyzer apparatus, and methods for automatically performing microscopic cell analysis tasks, such as counting and analyzing blood cells in biological samples. A small measured quantity of a biological sample, such as whole blood, is placed in a mixing bowl on the disposable test cartridge after being inserted into the cell analyzer. The analayzer also deposits a known amount of diluent/stain in the mixing bowl and mixes it with the blood. The analyzer takes a measured amount of the mixture and dispenses in a sample cup on the cartridge in fluid communication with an imaging chamber. The geometry of the imaging chamber is chosen to maintain the uniformity of the mixture, and to prevent cells from crowding or clumping as it is transferred into the imaging chamber by the analyzer. Images of all of the cellular components within the imaging chamber are counted and analyzed to obtain a complete blood count.
1. A method for analyzing and counting biological particles in a blood sample comprising:
a. separating a known amount of the blood sample;
b. diluting the known amount of blood sample with a known amount of diluent and/or stain;
c. mixing the known amount of blood sample and known amount of diluent and/or stain to obtain a substantially uniform mixture of sample and diluent and/or stain form of a liquid solution having a dilution ratio of between 10:1 and 250:1;
d. causing a known amount of the liquid mixture of sample and diluent and/or stain to flow into an imaging chamber of fixed dimensions defined inside of a test cartridge made with a transparent material, wherein the imaging chamber includes a bottom and sides and has a geometry that contains the mixture in such a way that the biological particles do not crowd or overlap as they settle to the bottom of the imaging chamber, and from which one or more digital images are captured that are at least statistically representative of a number and distribution of the biological particles in the blood sample; and
e. counting and analyzing at least one type of biological particle in the captured digital images with an automated microscope adapted to receive the test cartridge with the imaging chamber and utilizing bright field and florescent imaging of the liquid mixture in the imaging chamber, and with image processing and pattern recognition software.
2. A method of claim 1 further comprising displaying the one or more digital images of the particles.
3. A method of claim 1 wherein the counting and analyzing includes counting all of the particles in the imaging chamber.
4. A method of claim 1 wherein a rate of causing the mixture of diluent and/or stain and sample to flow into the imaging chamber is such that the mixture remains substantially uniform.
5. A method of claim 1 wherein a rate of causing the mixture of diluent and/or stain and sample to flow into the imaging chamber is about 2 uL per second.
6. A method of claim 1 wherein the mixing further includes mixing a cell sphering agent with the known amount of blood sample and known amount of diluent and/or stain to provide isovolumetric reshaping of red blood cells in the blood sample.
7. A method of claim 1 wherein the causing includes pulling the mixture into the imaging chamber by suction through a vacuum port in the test cartridge.
8. A method of claim 1 wherein the causing includes introducing the mixture into a sample cup which is in fluidic communication with the imaging chamber.
9. A method of claim 1 wherein the mixing mixes the sample and diluent and/or stain to obtain a mixture having a ratio of diluent and/or stain to sample of at least 10 to 1.
10. A method for analyzing and counting in claim 1 wherein the separating, the diluting, the mixing, and the causing are performed by a movable sampling probe mechanism.
11. A method of claim 1 wherein the automated microscope with image processing software counts all of white blood cells in the known amount of the mixture of sample and diluent and/or stain.
12. A method of claim 1 wherein the automated microscope with image processing software counts all of red blood cells in the known amount of the mixture of sample and diluent and/or stain.
13. A method of claim 1 wherein the automated microscope with image processing software performs a complete blood count.
14. A method of claim 1 wherein a width and depth of the imaging chamber is uniform and a length-to-width ratio of the imaging chamber is greater than 2 to 1.
15. A method of claim 1 wherein a width and depth of the imaging chamber are uniform and a length-to-width ratio of the imaging chamber is about 400 to 1.
16. A method of claim 1 wherein a width of the imaging chamber is uniform and between 0.5 mm and 2.5 mm.
17. A method of claim 1 wherein a depth and width of the imaging chamber are uniform and the width is 10 to 200 um.
18. A method of claim 1 wherein a depth of the imaging chamber is uniform and a shape of the imaging chamber in planar view is serpentine.
19. A method of claim 18 wherein an outside turning radius of the serpentine imaging chamber is about twice an inside turning radius of the serpentine imaging chamber.
20. A method of claim 1 wherein a depth of the imaging chamber is uniform and a shape of the imaging chamber in planar view is serpentine and having a width of 1.25 mm, an inside turning radius of 1.25 mm, an outside turning radius of 2.5 mm, and a depth of 0.125 mm.
21. A method of claim 1 wherein a shape of the imaging chamber in planar view is helical.
22. A method of claim 1 wherein a shape of the imaging chamber in planar view is castellated.