Method for determining a complete blood count on a white blood cell differential count
Systems and methods analyzing body fluids such as blood and bone marrow are disclosed. The systems and methods may utilize an improved technique for applying a monolayer of cells to a slide to generate a substantially uniform distribution of cells on the slide. Additionally aspects of the invention also relate to systems and methods for utilizing multi color microscopy for improving the quality of images captured by a light receiving device.
1. An automated process for analyzing a sample of fluid containing blood cells, the process comprising:
filling an applicator tip with a volume of the fluid containing blood cells;
positioning the applicator tip at a height above a slide, wherein the height is measured along a direction perpendicular to a surface of the slide;
dispensing a known volume of the fluid containing blood cells out of the applicator tip and while ejection of the fluid onto the slide is occurring, maintaining relative movement between the tip and the slide to lay down the entire known volume of fluid in two or more rows over a defined area of the slide, wherein the applicator tip height above the slide, a flow rate of the fluid out of the applicator tip, and a speed of the relative movement are selected such that the cells in the fluid settle onto the slide in a layer that is about one cell thick and such that morphology of the blood cells is sufficiently preserved to enable image-based cell analysis and calculation of a total cell count per microliter in the sample;
fixing and staining the blood cells on the slide;
illuminating the blood cells on the slide with a light source;
capturing at least one image of the blood cells corresponding to light in a first wavelength range of about 400 to 470 nm transmitted or reflected by the cells, and at least one image of the cells corresponding to light in a second wavelength range of about 470 to 750 nm transmitted or reflected by the cells;
automatically analyzing the images of the cells and calculating a total cell count per microliter in the sample of red blood cells, white blood cells, and platelets and determining a complete blood count (“CBC”) and a white blood cell (“WBC”) differential for the sample; and
displaying the images of the cells, the results of the CBC, and the results of the WBC differential in a graphical user interface (GUI) of a display, wherein the GUI is configured to allow an operator to interact with the images and sign off on the CBC results and the WBC differential results.
2. The process of claim 1 , wherein the two or more rows of cells form a gapless, contiguous monolayer of cells on a portion of the slide.
3. The process of claim 1 , wherein the fluid comprises a salt based or protein based diluent.
4. The process of claim 1 , wherein the entire known volume of fluid is dispensed onto the slide in four or more rows.
5. The process of claim 1 , wherein fixing and staining the cells on the slide comprises:
dispensing an amount of a fixative and an amount of a stain onto the slide with a dispenser; and
tilting the slide with a slide tilter to remove excess fixative and stain.
6. The process of claim 1 , wherein:
the height is less than about 110 microns;
the fluid is dispensed from the tip at a flow rate of about 0.04 to 0.1 microliters per second; and
the tip is translated relative to the slide at a velocity of about 10 to 100 mm per second.
7. The process of claim 1 , wherein illuminating the cells on the slide with a light source comprises:
generating illumination light from a light source for illuminating the cells on the slide;
applying a first light filter to the illumination light to produce filtered light corresponding to the first wavelength range of about 400 to 470 nm;
illuminating the cells with the filtered light corresponding to the first wavelength range;
applying a second light filter to the illumination light to produce filtered light corresponding to the second wavelength range of about 470 to 750 nm; and
illuminating the cells with the filtered light corresponding to the second wavelength range.
8. The process of claim 7 , further comprising:
applying a third light filter to the illumination light to produce filtered light corresponding to a third wavelength range; and
illuminating the cells with the filtered light corresponding to the third wavelength range, and capturing at least one image of the cells at the third wavelength range.
9. The process of claim 8 , wherein the first, second, and third wavelength ranges are selected from the range of 400 nm to 700 nm.
10. The process of claim 7 , wherein the first wavelength range is selected from a range of about 405 to 430 nm.
11. The process of claim 10 , further comprising determining a reticulocyte count from the at least one image of the cells at each of the first and second wavelength ranges.
12. The process of claim 10 , further comprising determining one or more of a red blood cell count and a platelet count from the at least one image of the cells at each of the first and second wavelength ranges.
13. The process of claim 7 , further comprising:
refining the at least one image of the cells at the first and second wavelength ranges by sharpening or compensating for spatial shifts or other distortions; and
combining two or more of the images to generate a multi-color image, wherein the images that are combined comprise at least one image at the first wavelength range and at least one image at the second wavelength range.
14. The process of claim 7 , further comprising determining spatial, densitometric, colorimetric and texture features of the cells for classification of a cell type.
15. The process of claim 7 , further comprising:
capturing a black and white image of the cells; and
correcting focus and image quality of the black and white image by adjusting a focal distance of a lens of a light receiving device while at least one filter is positioned between the slide and the light source.
16. The process of claim 1 , wherein illuminating the cells on the slide with a light source comprises:
illuminating the cells with light at the first wavelength range from a first light emitting diode (LED); and
illuminating the cells with light at the second wavelength range from a second LED.
17. The process of claim 16 , further comprising:
illuminating the cells with the light at a third wavelength range from a third LED, and capturing at least one image of the cells at the third wavelength range.
18. The process of claim 16 , wherein the first wavelength range is selected from a range of about 405 nm to 430 nm.
19. The process of claim 18 , further comprising determining a reticulocyte count from the at least one image of the cells at each of the first and second wavelength ranges.
20. The process of claim 18 , further comprising determining one or more of a red blood cell count and a platelet count from the at least one image of the cells at each of the first and second wavelength ranges.
21. The process of claim 1 , wherein the process does not include attaching a coverslip to the slide or using oil with an objective of a light receiving device.