Method for determining a complete blood count on a white blood cell differential count

Abstract

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.

It is claimed: 
     
         1 - 51 . (canceled) 
     
     
         52 . An automated system for analyzing a fluid containing blood cells, the system comprising:
 an applicator arranged to dispense a portion of the fluid onto a substrate;   an applicator controller arranged to regulate relative movement of the applicator with respect to the substrate or a substrate controller arranged to regulate relative movement of the substrate with respect to the applicator, or both an applicator controller and a substrate controller;   a light source arranged to emit light onto or through blood cells on the substrate;   a light receiving device arranged to capture light transmitted through or reflected by blood cells on the substrate;   a computer comprising one or more processors, wherein the computer is arranged to send or receive instructions and to send or receive information to or from the applicator controller, the substrate controller, the light source, and the light receiving device;   a display arranged to present information about the blood cells to an operator; and   software instructions stored on a storage device for controlling the system, wherein when the one or more processors execute the software instructions, the computer causes the system to:
 fill the applicator with a volume of the fluid containing blood cells; 
 position the applicator at a height above the substrate, wherein the height is measured along a direction perpendicular to a surface of the substrate; 
 dispense a known volume of the fluid containing blood cells out of the applicator and while ejection of the fluid onto the substrate is occurring, maintaining relative movement between the applicator and the substrate to lay down the entire known volume of fluid in two or more rows over a defined area of the substrate, wherein the applicator height above the substrate, a flow rate of the fluid out of the applicator, 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 fluid; 
 illuminate the blood cells on the slide with light from the light source; 
 capture at least one image of the blood cells corresponding to light in a first wavelength range of about 400 to 470 nm transmitted through or reflected by the cells, and at least one image of the blood cells corresponding to light in a second wavelength range of about 470 to 750 nm transmitted through or reflected by the cells; 
 analyze the images of the blood cells, calculate a total count of red blood cells, white blood cells, and platelets per microliter in the volume of fluid on the substrate, and determine a complete blood count (“CBC”) and a white blood cell (“WBC”) differential for the fluid; and 
 present the images of the blood cells, the results of the CBC, and the results of the WBC differential in a graphical user interface (GUI) on the display. 
   
     
     
         53 . The system of  claim 52 , further comprising
 a fixative dispenser arranged to apply a fixative to blood cells on the substrate; and   a stain dispenser arranged to apply a stain to the blood cells on the substrate;   wherein when the one or more processors execute the software instructions, the computer causes the system to fix and stain the blood cells on the substrate.   
     
     
         54 . The system of  claim 53 , further comprising a slide tilter arranged to remove excess fixative or stain or both from the substrate. 
     
     
         55 . The system of  claim 52 , 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. 
     
     
         56 . The system of  claim 52 , further comprising a gas moving device arranged to move air over the substrate to dry the fluid. 
     
     
         57 . The system of  claim 52 , wherein the applicator controller and the substrate controller are configured as one component of the system. 
     
     
         58 . The system of  claim 52 , wherein when the one or more processors execute the software instructions, the computer causes the system to further determine a hematocrit level or a hemoglobin level, or both. 
     
     
         59 . The system of  claim 52 , further comprising a first reservoir for storing the fluid, a second reservoir for storing a diluent, and a mixer for mixing the fluid with the diluent to form a diluted fluid to be dispensed onto the substrate. 
     
     
         60 . The system of  claim 52 , wherein when the one or more processors execute the software instructions, the computer causes the system to
 refine the images of the blood cells by compensating for spatial shifts or other distortions; and   combine two or more black and white images captured by the light receiving device to generate a multi-color image for presentation on the display.   
     
     
         61 . The system of  claim 52 , wherein when the one or more processors execute the software instructions, the computer causes the system to determine spatial, densitometric, colorimetric, and texture features of the blood cells for classification of cell type. 
     
     
         62 . The system of  claim 52 , wherein the two or more rows are formed by the applicator tip dispensing the fluid in a first row in a first direction and then quickly turning and moving in a second direction opposite the first direction and in parallel to the first row to lay down a second row adjacent the first row. 
     
     
         63 . The system of  claim 52 , wherein the applicator controller is instructed to
 adjust the height of the applicator above the substrate to be less than about 110 microns;   eject the fluid out of the applicator at a flow rate of less than about 0.1 microliters per second; and   regulate the speed of the relative movement between the applicator and substrate to be about 10 to 100 mm per second.   
     
     
         64 . The system of  claim 52 , wherein the light source comprises:
 a first light filter to produce filtered light corresponding to the first wavelength range of about 400 to 470 nm; and   a second light filter to produce filtered light corresponding to the second wavelength range of about 470 to 750 nm.   
     
     
         65 . The system of  claim 64 , wherein the light source further comprises a third light filter to produce filtered light corresponding to a third wavelength range. 
     
     
         66 . The system of  claim 52 , wherein the first wavelength range is selected from a range of about 400 to 430 nm. 
     
     
         67 . The system of  claim 52 , wherein the light source comprises:
 a first light emitting diode (LED) that emits light at the first wavelength range; and   a second LED that emits light at the second wavelength range.   
     
     
         68 . The system of  claim 67 , wherein the light source further comprises a third LED that emits light corresponding to a third wavelength range. 
     
     
         69 . The system of  claim 52 , wherein when the one or more processors execute the software instructions, the computer causes the system to further determine one or more of a red blood cell count and a platelet count from the at least one image of the blood cells at each of the first and second wavelength ranges. 
     
     
         70 . The system of  claim 52 , wherein when the one or more processors execute the software instructions, the computer causes the system to further determine a reticulocyte count from the at least one image of the blood cells at each of the first and second wavelength ranges.