Accelerated Wright-Giemsa and May-Grunwald Staining Methods

Abstract

The present disclosure provides methods for carrying out Romanowsky-type stains, specifically Wright-Giemsa and May-Grünwald stains, quickly and efficiently. The methods greatly reduce the overall amount of time required to complete a Wright-Giemsa stain or a May-Grünwald stain of sufficient quality on a biological sample. The subject methods can be applied to both manual and automated staining procedures.

1 .- 58 . (canceled) 
     
     
         59 . A computer-implemented method for executing instructions stored on a non-transitory computer-readable medium for performing a Wright-Giemsa or May-Grunwald stain on one or more biological samples in an automated staining method, comprising the following steps performed by one or more computer processors:
 applying the biological sample to a substrate via a sample application subsystem;   contacting the sample with a fixation reagent contained in a fixation reagent bath by transferring the sample from the sample application subsystem to the fixation reagent bath via a sample transfer subsystem;   contacting the sample with a Wright-Giemsa staining reagent or the May-Grunwald staining reagent contained in a Giemsa staining reagent bath or a May-Grunwald staining reagent bath by transferring the sample from the fixation reagent bath to the Giemsa staining reagent bath or the May-Grunwald staining reagent bath via the sample transfer subsystem; and   contacting the sample with one or more rinse reagents contained in one or more rinse reagent baths by transferring the sample from the Giemsa staining reagent bath or the May-Grunwald staining reagent bath to the one or more rinse reagent baths via the sample transfer subsystem.   
     
     
         60 . The method of  claim 59 , comprising contacting the sample with the fixation reagent for about 20 seconds up to about 40 seconds. 
     
     
         61 . The method of  claim 60 , wherein the fixation reagent is methanol. 
     
     
         62 . The method of  claim 59 , comprising contacting the sample with the Wright-Giemsa staining reagent for about 20 seconds up to about 50 seconds. 
     
     
         63 . The method of  claim 59 , wherein the Wright-Giemsa staining reagent comprises methanol. 
     
     
         64 . The method of  claim 59 , comprising contacting the sample with the May-Grunwald staining reagent for about 165 seconds up to about 195 seconds. 
     
     
         65 . The method of  claim 59 , wherein the May-Grunwald staining reagent comprises methanol. 
     
     
         66 . The method of  claim 59 , comprising contacting the sample with a first rinse reagent from the one or more rinse reagents for about 105 seconds up to about 135 seconds. 
     
     
         67 . The method of  claim 66 , wherein the first rinse reagent from the one or more rinse reagents is a phosphate buffer. 
     
     
         68 . The method of  claim 67 , wherein the phosphate buffer is a high salt phosphate buffer. 
     
     
         69 . The method of  claim 67 , wherein the phosphate buffer is a low salt phosphate buffer. 
     
     
         70 . The method of  claim 67 , wherein the phosphate buffer has a pH of about 5.0 up to about 9.0. 
     
     
         71 . The method of  claim 59 , comprising contacting the sample with a second rinse reagent from the one or more rinse reagents for about 5 seconds up to about 30 seconds. 
     
     
         72 . The method of  claim 71 , wherein the second rinse reagent is a de-ionized water. 
     
     
         73 . The method of  claim 59 , further comprising placing the sample in a drying chamber. 
     
     
         74 . The method of  claim 59 , wherein the substrate is a glass microscope slide. 
     
     
         75 . The method of  claim 59 , wherein the sample is a biological fluid. 
     
     
         76 . The method of  claim 75 , wherein the sample is a blood sample. 
     
     
         77 . The method of  claim 59 , wherein the sample is a bone marrow sample. 
     
     
         78 . The method of  claim 59 , wherein the concentration of each reagent remains constant during the period of time that the sample is contacted with the reagent. 
     
     
         79 . The method of  claim 59 , wherein the substrate remains motionless for the period of time during which the sample is contacted with any reagent.