The Fixation of Tissues Vitally Stained with Trypan Blue |
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Authors: | Duncan C Hetherington Edna H Tompkins |
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Institution: |
a Department of Anatomy, Vanderbilt University School of Medicine, Nashville, Tennessee |
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Abstract: | The following fixative is recommended for tissues vitally stained with trypan blue: Chloroform, 2 parts; absolute ethyl alcohol, 2 parts; glacial acetic acid, 1 part; mercuric chloride to the point of saturation.
The tissue should be fixed 1 to 2 hours; transferred to 95% ethyl alcohol for 12 hours; to absolute alcohol for 12 to 24 hours; to a mixture of absolute alcohol and xylol for 1/2 hour, and finally to xylol, before embedding in paraffin. Cedar oil may be used for clearing in the place of xylol; in that case the tissues should be transferred from absolute alcohol to a mixture of absolute alcohol and cedar oil for 24 hours before placing in cedar oil alone.
Various counterstains can be used; Mayer's carmalum is excellent. |
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