Composition for high stringency cell treatment and antigen retrieval

Abstract

The present invention relates to a cell treatment composition for the permeabilization of fixed blood cells, to the use of said composition, to a method for the treatment of a biological sample comprising fixation of said sample and subsequently contacting said biological sample with said cell treatment composition. The invention further relates and to a kit comprising said cell treatment composition.

What is claimed is: 
     
         1 . A method for treating an isolated biological sample, said sample comprising at least white blood cells, comprising the steps of:
 (a) a fixation step comprising contacting said biological sample with a fixative, wherein said fixative is added in a sufficient amount to achieve at least partial cross-linking of proteins, lipoproteins and nucleic acid molecules; and   (b) a permeabilization step comprising contacting said fixed biological sample with the cell treatment composition according to  claim 1 .   
     
     
         2 . The method according to  claim 1 , further comprising
 (c) a labeling step comprising contacting said permeabilized biological sample with at least one detectably labeled binding agent specific for an intracellular and/or an extracellular epitope.   
     
     
         3 . The method according to  claim 1 , said method comprising incubating said biological sample with said fixative for 5 to 15 minutes at a temperature of between 15° C. and 30° C. 
     
     
         4 . The method according to  claim 1 , said method comprising incubating said fixed biological sample with said cell treatment composition for 2 to 10 minutes at a temperature of between 20° C. and 50° C. 
     
     
         5 . The method according to  claim 1 , said method comprising washing the permeabilized biological sample. 
     
     
         6 . The method according to  claim 1 , wherein said cell treatment composition permeabilizes said white blood cells and unmasks intracellular epitopes of said white blood cells. 
     
     
         7 . The method according to  claim 6 , wherein said intracellular epitopes comprise phospho-epitopes. 
     
     
         8 . The method according to  claim 1 , said biological sample comprising whole blood, bone marrow, or an isolated subpopulation of leukocytes. 
     
     
         9 . The method according to  claim 1 , said white blood cells comprising lymphocytes or monocytes.