Basophil Analysis System and Method

Abstract

Provided herein are systems and methods for analyzing blood samples, and more specifically for performing a basophil analysis. In one embodiment, the systems and methods include: (a) staining a blood sample with an exclusive cell membrane permeable fluorescent dye; and then (b) using measurements of light scatter and fluorescence emission to distinguish basophils from other WBC sub-populations. In one embodiment, the systems and methods include performing a basophil cluster analysis of the blood sample, based on the combination of light scatter and fluorescence measurements.

1 - 19 . (canceled) 
     
     
         20 . A method of performing a basophil analysis with an automated hematology analyzer, the method comprising:
 (a) staining a sample of whole blood with a cell membrane permeable, nucleic acid binding fluorescent dye;   (b) exciting the sample from step (a) with an excitation source as the sample traverses a flow cell in the hematology analyzer;   (c) collecting a plurality of light scatter signals and a fluorescence emission signal from the excited sample;   (d) prior to performing a basophil cluster analysis, excluding nuclei-free particles and retaining nuclei-containing particles using only a fluorescence trigger configured in the hematology analyzer and that is limited to fluorescence emission signals and is set to a fluorescence magnitude that is greater than fluorescence emission signals from RBCs, including RBC fragments, and is less than fluorescence emission signals from white blood cells (WBCs); and   (e) performing the basophil cluster analysis on the nuclei-containing particles collected in step (d).   
     
     
         21 . The method of  claim 20 , wherein the basophil cluster analysis includes a coarse clustering of the nuclei-containing particles. 
     
     
         22 . The method of  claim 21 , wherein the basophil cluster analysis further includes a fine clustering of the nuclei-containing particles, utilizing a multi-dimensional, probability distance metric as a determinant to combine similar clusters. 
     
     
         23 . (canceled) 
     
     
         24 . The method of  claim 20 , wherein the excitation source has a wavelength of from about 350 nm to about 700 nm. 
     
     
         25 . The method of  claim 20 , wherein the excitation source has a wavelength of about 488 nm. 
     
     
         26 . The method of  claim 20 , wherein the plurality of light scatter signals include: (1) axial light loss, (2) intermediate angle scatter, and (3) 90° side scatter. 
     
     
         27 . The method of  claim 20 , wherein the plurality of light scatter signals include: (1) axial light loss, (2) intermediate angle scatter, (3) 90° polarized side scatter, and (4) 90° depolarized side scatter. 
     
     
         28 . (canceled) 
     
     
         29 . The method of  claim 20 , wherein performing the basophil cluster analysis comprises distinguishing basophils within the blood sample from neutrophils, monocytes, and eosinophils within the blood sample based on the measurements of (1) axial light loss, (2) intermediate angle scatter, (3) 90° side scatter, and (4) fluorescence. 
     
     
         30 . The method according to  claim 20 , wherein the excitation source is a laser. 
     
     
         31 . The method according to  claim 20 , further comprising incubating the blood sample of step (a) for an incubation period of time. 
     
     
         32 . The method according to  claim 31 , wherein the incubation period of time is less than 25 seconds. 
     
     
         33 . The method according to  claim 31 , wherein the incubation period of time is less than 17 seconds. 
     
     
         34 . The method according to  claim 31 , wherein the incubation period of time is less than 9 seconds. 
     
     
         35 . The method according to  claim 31 , wherein the blood sample is incubated at a temperature ranging from 30° C. to 50° C. 
     
     
         36 . The method according to  claim 31 , wherein the blood sample is incubated at a temperature of about 40° C. 
     
     
         37 . The method according to  claim 20 , wherein a single fluorescent dye is used to identify, quantify, and analyze nRBCs and a plurality of WBC subpopulations at once.