A one-solution triacid general stain which differentiates oxygenated from non-oxygenated red blood corpuscles

H.F. Steedman

Research output: Contribution to journalArticle

Abstract

Tissues from representative mammals, amphibia and invertebrates were fixed for 5-24 hr in either an aqueous solution of 8% p-toluene sulfonic acid (PTSA) or in 10% formalin to which 5 gm PTSA/100 ml had been added, and processed through embedding in polyethylene glycol 400 distearate in the usual manner. Sections cut at 4-6 μ were floated on 0.2% gelatin containing 1.25% formalin, and spread and dried on slides at a temperature not exceeding 25 C. Wax was removed with xylene, and the sections brought to water through ethanol as usual. The working staining solution was made from three stock solutions: A. Chlorantine fast blue 2RLL, 0.5%; B. Cibacron turquoise blue G-E, 0.5%; C. Procion red M-P, 0.5%-each of which was dissolved in 98.5 ml of distilled water to which 0.5 ml of glacial acetic acid and 0.5 ml of propylene glycol monophenyl ether (a fungicide) had been added. For use, the three solutions were mixed in the proportions: A, 3; B, 4; and C, 3 volumes. Staining time was uncritical, 10-30 min usually sufficing for 6 μ sections. The chief feature of the staining is the differentiation of oxygenated and nonoxygenated red blood corpuscles, in reds and blues respectively. Connective tissue stained blue or blue-green and mucin, green. Nuclei and cytoplasm stain according to their condition at the time of fixation. The mixed stain keeps well, remaining active after 2 yr of storage.
LanguageEnglish
Pages247-253
Number of pages7
JournalBiotechnic and Histochemistry
Volume45
Issue number6
DOIs
StatusPublished - 1970

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Coloring Agents
Erythrocytes
Staining and Labeling
Formaldehyde
Xylenes
Propylene Glycol
Water
Waxes
Amphibians
Mucins
Invertebrates
Gelatin
Acetic Acid
Ether
Connective Tissue
Mammals
Cytoplasm
Ethanol
Temperature
4-toluenesulfonic acid

Cite this

A one-solution triacid general stain which differentiates oxygenated from non-oxygenated red blood corpuscles. / Steedman, H.F.

In: Biotechnic and Histochemistry, Vol. 45, No. 6, 1970, p. 247-253.

Research output: Contribution to journalArticle

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