Summary
Crystal violet from Gram stained S. aureus can be extracted completely by 95% ethanol if the stained bacteria is pre-treated with dilute sodium thiosulphate solution. Thiosulphate removes iodine form the cell component-dye-iodine complex instantaneously and renders the dye extractable by the differentiating medium. 1∶1 alcoholic solutions of aniline, dimethyl aniline, nitro-benzene, benzene, toluene or xylol can also extract the color from the stained S. aureus; the extraction with the first three solvents is almost exhaustive while with the latter solvents extraction is appreciable but incomplete. These solvents can form charge-transfer complexes with iodine. The findings indicate that the stability of the cell component-dye-iodine complex determines the Gram-character of the cell. A model hasbeen presented for the Gram cell component-dye-iodine complex.
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Basu, P.S., Biswas, B.B. & Pal, M.K. Extraction of crystal violet-iodine complex from gram positive bacteria by different solvents and its implication on gram differentiation. Histochemie 14, 221–229 (1968). https://doi.org/10.1007/BF00306318
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DOI: https://doi.org/10.1007/BF00306318