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The addition of buffer mixtures to toluidine blue O, thionin and new methylene blue N improves their use in the staining of mucus after formaldehyde fixation. Overstaining is minimi The buffer mixtures used consisted of variable proportions of M/10 citric acid and M/5 anhydrous Na2HPO4 in 25% methanol. Connective tissue mucus stained satisfactorily with these dyes at a buffer pH range of 3.4 and 3.95 and epithelial mucus at a range of 2.2 to 3.95. The corresponding ratios of M/10 citric acid: M/5 Na2HPO4 are 16:4 to 14:6 and 20 A to 14:6 respectively.
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Benjamin Highman (1945) studied this question.
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