Reagent, reagent kit and analyzing method

Abstract

A method for analyzing platelet is described. In the method, a measurement sample is prepared by mixing a sample and a dye for staining platelet. The dye is selected from the group consisting of Capri blue, Nile blue and brilliant cresyl blue. By irradiating cells in the measurement sample with light, scattered light and fluorescence emitted from the cells is measured. The platelet is detected on the basis of the scattered light and the fluorescence. A reagent kite and a reagent are also described.

1 . A method for analyzing platelet, comprising the steps of: 
 preparing a measurement sample by mixing a sample and a dye for staining platelet, the dye being selected from the group consisting of Capri blue, Nile blue and brilliant cresyl blue;    measuring scattered light and fluorescence emitted from cells in the measurement sample by irradiating the cells with light; and    detecting platelet on the basis of the scattered light and the fluorescence.    
     
     
         2 . The method according to  claim 1 , wherein the preparing step comprises mixing the first dye for staining platelet, the sample and a second dye for staining reticulocyte to prepare the measurement sample, and the detecting step comprises detecting platelet and reticulocyte on the basis of the measured scattered light and the measured fluorescence.  
     
     
         3 . The method according to  claim 1 , wherein the measurement sample is introduced into a flow cell of a flow cytometer, and cells in the measurement sample flowing in the flow cell are irradiated with light.  
     
     
         4 . The method according to  claim 1 , further comprising a step of counting the detected platelet.  
     
     
         5 . The method according to  claim 2 , further comprising a step of counting the detected platelet and a step of counting the detected reticulocyte.  
     
     
         6 . The reagent according to  claim 2 , wherein the second dye is a cyanine dye capable of staining a nucleic acid.  
     
     
         7 . A reagent kit for analyzing platelet, comprising: 
 a first reagent containing a buffer; and    a second reagent containing a dye for staining platelet, wherein the dye is selected from the group consisting of Capri blue, Nile blue and brilliant cresyl blue.    
     
     
         8 . The reagent kit according to  claim 7 , wherein the second reagent comprises a second dye for staining reticulocyte, and the reagent kit is capable of analyzing platelet and reticulocyte.  
     
     
         9 . The reagent kit according to  claim 7 , further comprising a third reagent comprising a second dye for staining reticulocyte, wherein the reagent kit is capable of analyzing platelet and reticulocyte.  
     
     
         10 . The reagent kit according to  claim 8 , wherein the second dye is a cyanine dye capable of staining a nucleic acid.  
     
     
         11 . The reagent kit according to  claim 10 , wherein the second dye is represented by the following formula (I):  
       
         
           
           
               
               
           
         
       
       wherein: 
 R 1  represents a hydrogen atom, a C1 to C6 alkyl group, or —CH 2 (CHR 5 ) x OR 6 ;  
 R 2  and R 3  independently represent a hydrogen atom, a halogen atom, a cyano group, a C1 to C6 alkyl group, a C1 to C6 alkoxy group, an aryl group, or an aralkyl group;  
 R 4  represents a C1 to C6 alkyl group, —CH 2 (CHR 7 ) y OR 8 , an aryl group, or an aralkyl group;  
 R 5  and R 7  independently represent a hydrogen atom or a C1 to C3 hydroxyalkyl group;  
 R 6  and R 8  independently represent a hydrogen atom, an acyl group, or a C1 to C3 alkyl group;  
 Z represents a sulfur atom, an oxygen atom, a selenium atom, or CR 9 R 10 ;  
 R 9  and R 10  independently represent a C1 to C3 alkyl group;  
 n is an integer of 1 or 2;  
 x and y independently represent an integer of 0 to 3; and  
 X −  is an anion.  
 
     
     
         12 . The reagent kit according to  claim 8 , wherein at least one of the first reagent or the second reagent comprises a multivalent anion for suppressing nonspecific staining of red blood cell.  
     
     
         13 . The reagent kit according to  claim 7 , wherein the second reagent has a pH of 6.0 to 11.0.  
     
     
         14 . The reagent kit according to  claim 7 , wherein the second reagent has an osmotic pressure of 150 to 600 mOsm/kg.  
     
     
         15 . The reagent kit according to  claim 7 , wherein at least one of the first reagent or the second reagent comprises a staining promoter for promoting penetration of the dye.  
     
     
         16 . The reagent kit according to  claim 15 , wherein the staining promoter is a cationic surfactant.  
     
     
         17 . The reagent kit according to  claim 7 , wherein the dye for staining platelet stains platelets so as to be distinguishable from lipid particles.  
     
     
         18 . The reagent kit according to  claim 7 , wherein the dye for staining platelet stains platelet so as to be distinguishable from fragmented red blood cells.  
     
     
         19 . A reagent for analyzing platelet by flow cytometer, comprising a dye for staining platelet, wherein the dye is selected from the group consisting of Capri blue, Nile blue and brilliant cresyl blue.  
     
     
         20 . The reagent according to  claim 19 , further comprising a second dye for staining reticulocyte, wherein the reagent is capable of analyzing platelet and reticulocyte.