Serial Sectioning of Insects with Hard Exoskeleton by Dissolution of the Exocuticle
作者:
HaasFabian,
期刊:
Biotechnic&Histochemistry
(Taylor Available online 1992)
卷期:
Volume 67,
issue 1
页码: 50-54
ISSN:1052-0295
年代: 1992
DOI:10.3109/10520299209110006
出版商: Taylor&Francis
关键词: serial sectioning;insects;cuticle dissolution;paraffin embedding
数据来源: Taylor
摘要:
The method reported here was designed to produce paraffin serial sections as thin as 5 Mm of insects or other arthropods with a hard cuticle. Heads and abdomens ofApis mellifera, Eristalomyia tenaxandTenebrio molitorwere fixed with Schaffer's liquid, dehydrated with 80% ethanol, 90% ethanol, two changes of 100% isopropanol (2 hr each) and 12 hr in a 1:1 mixture of paraffin (58 C melting point) at 60 C. They were molded in paraffin after 12 hr of infiltration under a partial vacuum at 60 C. Large body openings of objects were sealed with paraffin to prevent infiltration of solvents.Thereafter, the outer paraffin was removed manually and with xylene (15 min); the cuticle was rehydrated with 100% isopropanol and 100% ethanol (15 min each). The objects were then treated with Sputofluol (Merck; a mixture of NaOH and NaCIO) until they became white or their colorless endocuticle was stainable with aniline blue WS (C.I. 42755) after rinsing in a 50% acetic acid solution (v/v). They were then dehydrated with 100% ethanol and 100% isopropanol (15 min each) and subsequently re-embedded in paraffin. They were molded, sectioned, stained and mounted as usual.
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