White blood cell differentiation reagent and method

Abstract

A reagent for four-part differentiation of white blood cells is provided. In one embodiment the reagent has an osmolarity below 50 mOsm/kg H 2 O. A method for differentiating white blood cells using the reagent is also provided. The disclosure provides for a rapid lysis of red blood cells and four-part differentiation of white blood cells. The reagent may be simple in components and a surfactant is not necessary, but optional. A wide range of pH values may be suitable for the reagent.

1 . A reagent for differentiation of white blood cells, the reagent comprising a composition configured for four-part differentiation of white blood cells into four groups, including three groups corresponding to lymphocytes, monocytes, and eosinophil granulocytes and one group corresponding to neutrophil and basophil granulocytes, wherein the composition has an osmolarity below 50 mOsm/kg H 2 O. 
   
   
       2 . The reagent according to  claim 1 , wherein the composition has a pH value between 4-12. 
   
   
       3 . The reagent according to  claim 2 , wherein the composition comprises water as a solvent and at least one substance selected from the following: i) an inorganic salt, and ii) a buffer. 
   
   
       4 . The reagent according to  claim 3 , wherein the composition comprises an inorganic salt and the inorganic salt is selected from at least one of the following: NaCl, KCl, LiCl, Na 2 SO 4  and K 2 SO 4 . 
   
   
       5 . The reagent according to  claim 2 , wherein the composition comprises water as a solvent and at least one substance selected from of the following: i) an inorganic salt, ii) a surfactant, iii) an organic compound having a hydrophobic group and an anionic group, and iv) a buffer. 
   
   
       6 . The reagent according to  claim 5 , wherein the composition comprises an inorganic salt and the inorganic salt is selected from at least one of the following: NaCl, KCl, LiCl, Na 2 SO 4  and K 2 SO 4 . 
   
   
       7 . The reagent according to  claim 5 , wherein the composition comprises a surfactant and the surfactant is configured to lyse red blood cells and partly damage membranes of white blood cells, and the surfactant is chosen from at least one of: a cationic surfactant, an anionic surfactant or a zwitterionic surfactant. 
   
   
       8 . The reagent according to  claim 5 , wherein the composition comprises an organic compound and the organic compound is configured to bind with cations in white blood cells to engender a morphological difference among white blood cells; the hydrophobic group of the organic compound is selected from at least one: of aromatic groups, hydrocarbonyl groups having more than six carbon atoms or heterocyclyl groups having more than six carbon atoms. 
   
   
       9 . The reagent according to  claim 5 , wherein the composition comprises an organic compound and the organic compound is chosen from at least one of: an acidic pigment, terephthalic acid, phthalic acid, chromotropic acid or naphthalenesulfonic acid, or a derivative or a salt of terephthalic acid, phthalic acid, chromotropic acid or naphthalenesulfonic acid. 
   
   
       10 . The reagent according to  claim 5 , wherein the composition comprises a surfactant and an organic compound, the surfactant is decyltrimethylammonium chloride and the organic compound is Bromocresol Green. 
   
   
       11 . A method for differentiation of white blood cells, wherein the method comprises:
 obtaining a reagent for differentiation of white blood cells according to  claim 1 ;   mixing the reagent with a whole blood sample; and   detecting information regarding size and morphology of the white blood cells from the whole blood sample after mixing with the reagent.   
   
   
       12 . The method according to  claim 11 , wherein mixing the reagent comprises mixing the reagent with a whole blood sample in a ratio of 50:1 to 200:1 under a temperature between ambient temperature and 40° C. for between 10-20 seconds; and detecting information comprises detecting information regarding size and morphology of the white blood cells through low-angle scattered light and high-angle scattered light respectively, wherein a detection angle for the low-angle scattered light is between 2-5 degrees and a detection angle for the high-angle scattered light is between 8-20 degrees. 
   
   
       13 . The method according to  claim 11 , wherein obtaining the reagent comprises obtaining a reagent comprising a composition with pH value between 4-12. 
   
   
       14 . The method according to  claim 13 , wherein obtaining the reagent comprises obtaining a reagent comprising a composition with water as a solvent and at least one substance selected from the following: i) an inorganic salt, and ii) a buffer. 
   
   
       15 . The method according to  claim 14 , wherein the composition comprises an inorganic salt and the inorganic salt is selected from at least one of the following: NaCl, KCl, LiCl, Na 2 SO 4  and K 2 SO 4 . 
   
   
       16 . The method according to  claim 13 , wherein obtaining the reagent comprises obtaining a reagent comprising a composition with water as a solvent and at least one substance selected from of the following: i) an inorganic salt, ii) a surfactant, iii) an organic compound having a hydrophobic group and an anionic group, and iv) a buffer. 
   
   
       17 . The method according to  claim 16 , wherein the composition comprises an inorganic salt and the inorganic salt is selected from at least one of the following: NaCl, KCl, LiCl, Na 2 SO 4  and K 2 SO 4 . 
   
   
       18 . The method according to  claim 16 , wherein the composition comprises a surfactant and the surfactant is configured to lyse red blood cells and partly damage membranes of white blood cells, and the surfactant is chosen from at least one of: a cationic surfactant, an anionic surfactant or a zwitterionic surfactant. 
   
   
       19 . The method according to  claim 16 , wherein the composition comprises an organic compound and the organic compound is configured to bind with cations in white blood cells to engender a morphological difference among white blood cells; the hydrophobic group of the organic compound is selected from at least one: of aromatic groups, hydrocarbonyl groups having more than six carbon atoms or heterocyclyl groups having more than six carbon atoms. 
   
   
       20 . The method according to  claim 16 , wherein the composition comprises an organic compound and the organic compound is chosen from at least one of: an acidic pigment, terephthalic acid, phthalic acid, chromotropic acid or naphthalenesulfonic acid, or a derivative or a salt of terephthalic acid, phthalic acid, chromotropic acid or naphthalenesulfonic acid. 
   
   
       21 . The method according to  claim 16 , wherein the composition comprises a surfactant and an organic compound, the surfactant is decyltrimethylammonium chloride and the organic compound is Bromocresol Green.