Cytological staining compositions and uses thereof

Abstract

The present subject matter is directed to a composition for staining cytological material comprising a cationic dye component, a hygroscopic polyol and optionally a water-soluble solvent, water or water-miscible solvent, methods of use of the compositions. Advantageously, the present compositions do not require the use of a cover slide as is required in known staining fixatives. The compositions are able to retain cell morphology for a period of time such that a cover slide is not required. Further, the compositions do not contain hazardous levels of organic components. Preferably, the compositions consist essentially of Azure C, glycerol and optionally water. In another aspect, the present subject matter is directed to a method of characterizing a cell sample comprising contacting the cell sample with a composition comprising a dye component and a hygroscopic polyol and subjecting the sample to analysis to determine the presence or absence of abnormal cells.

1 .- 28 . (canceled) 
     
     
         29 . A method of preparing a cytological preparation comprising: contacting a cell sample with a cytological staining composition, thereby staining cells in the cell sample, wherein the cytological staining composition comprises:
 i. a dye component comprising one or more water-soluble, pH neutral, cationic dyes; and   ii. one or more hygroscopic polyols present at less than about 10% v/v of the total composition.   
     
     
         30 . The method of  claim 29 , wherein cytological staining composition comprises water, a water-soluble solvent, a water-miscible solvent, or mixtures thereof. 
     
     
         31 . The method of  claim 29 , wherein the cell sample is associated with a substrate. 
     
     
         32 . The method of  claim 31 , wherein said substrate is selected from the group consisting of a microscope slide, a cuvette, a microtiter tray, a cell culture tray and a coverslip. 
     
     
         33 . The method of  claim 31 , the method does not comprise associating a solid cover slide with said sample associated with said substrate. 
     
     
         34 . The method of  claim 29 , wherein the cell sample comprises cervical cells, abnormal cells, tumor cells, cells obtained in tissue aspiration, cells obtained in lavage collection, cells obtained in biopsy, or mixtures thereof. 
     
     
         35 . A method of characterizing a cytological sample comprising:
 a. contacting a cell sample with a cytological staining composition, wherein said sample is associated with a substrate, and   b. subjecting said sample to an assay to determine the presence or absence of abnormal cells,   wherein the cytological staining composition comprises:
 i. a dye component comprising one or more water-soluble, pH neutral, cationic dyes; and 
 ii. one or more hygroscopic polyols present at less than about 10% v/v of the total composition. 
   
     
     
         36 . The method of  claim 35 , wherein the cytological staining composition comprises water, a water-soluble solvent, a water-miscible solvent, or mixtures thereof. 
     
     
         37 . The method of  claim 35 , wherein the abnormal cells comprise a cancer cell. 
     
     
         38 . The method of  claim 37 , wherein the cancer is selected from the group consisting of cervical, cancers of the reproductive system, brain, lung, liver, spleen, kidney, renal cell and renal pelvis cancers, lymph node, small intestine, pancreas, blood cells, bone, colon/colorectal, stomach, breast, endometrium, prostate, testicle, ovary, central nervous system, skin, head and neck, thyroid, para-thyroid, esophagus, bone marrow, hematological cancers, leukemia, acute promyelocytic leukemia, lymphoma, multiple myeloma, myelodysplasia, and myeloproliferative disease. 
     
     
         39 . The method of  claim 37 , wherein the cancer cell is a cervical cancer cell. 
     
     
         40 . A composition for cytology staining, comprising:
 i. Azure C;   ii. one or more hygroscopic polyol(s), said polyol(s) present at less than about 10% v/v of the total composition; and optionally   iii. water, a water-soluble solvent or water-miscible solvent, or mixtures thereof.   
     
     
         41 . The composition of  claim 40 , comprising water, a water-soluble solvent, a water-miscible solvent, or mixtures thereof. 
     
     
         42 . The composition of  claim 40 , wherein Azure C is present in an amount of about 0.01 to about 0.0001% w/v. 
     
     
         43 . The composition of  claim 40 , wherein said hygroscopic polyol is selected from the group consisting of glycerol, ethylene glycol, diethylene glycol, triethylene glycol, propylene glycol, dipropylene glycol, tripropylene glycol, 1,3-butanediol, 2,3-butanediol, 1,4-butanediol, 3-methyl-1,3-butanediol, 1,5-pentanediol, tetraethylene glycol, 1,6-hexanediol, 2-methyl-2,4-pentanediol, polyethylene glycol, 1,2,4-butanetriol, 1,2,6-hexanetriol, 2-ethyl-2-methyl-1,3-propanediol, 3,3-dimethyl-1,2-butanediol, 2,2-diethyl-1,3-propanediol, 2-methyl-2-propyl-1,3-propanediol, 2,4-dimethyl-2,4-pentanediol, 2,5-dimethyl-2,5-hexanediol, 5-hexene-1,2-diol, 2-ethyl-1,3-hexanediol, 2,2,4-trimethyl-1,3-pentanediol, butylene glycol, pentaerthyritol, mannitol, sorbitol, di-glycerol, tri-glycerol, tetra-glycerol, erythritol, 2-hydroxymethyl-2-methyl-1,3-propanediol (trimethylolethane), 2-ethyl-2-(hydroxymethyl)-1,3-propanediol (trimethylolpropane), and 1,2,4-hexanetriol and mixtures thereof. 
     
     
         44 . The composition of  claim 40 , wherein the hygroscopic polyol is present in an amount of less than 5% v/v. 
     
     
         45 . The composition of  claim 40 , wherein the solvent is a non-flammable, biocompatible solvent. 
     
     
         46 . The composition of  claim 40 , wherein said solvent is water. 
     
     
         47 . A composition for cytology staining, comprising:
 i. dye component consisting of Azure C;   ii. glycerol, wherein said glycerol is present in an amount less than about 10% v/v of said composition; and optionally   iii. water.   
     
     
         48 . The composition of  claim 47 , wherein
 i. Azure C is present in an amount of about 0.004% w/v; and   ii. glycerol is present in an amount of about 3% v/v of said composition.