A Simple Toluidine Blue—Basic Fuchsin Stain for Spermatozoa in Epoxy Sections

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Abstract
Differential staining of cell components of spermatozoa is readily accomplished in Epon or Araldite sections 0.5-1 u thick from rat and hamster testis and epididymis, and stained as follows; 1% aqueous toluidine blue buffered at pH 6, 0.5-3 min. at 90 C; washed in distilled water; 1% basic fuchsin in 50% alcohol, 3-5 min. at 20-25 C; differentiated with 70% alcohol; allowed to dry; and mounted in a resin of high refraction (DPX was used). The staining results were acrosome, bright magenta; nucleus, deep blue; mitochondrial sheath of the middle-piece, pinkish purple; and tail, pale red. This procedure combined with staining of collagen by applying 2% aqueous phospho-tungstic acid 1-2 min. as a mordant, followed by 1% light green in 50% alcohol containing 1% acetic acid, 1-2 min. at 20-25 C, gives polychromatic staining and is useful as a general stain for other epoxy-embedded tissues.