Novel applications of acrylamide for cryosectioning of isolated cells, tissues, and arthropods.
Cross-linked and uncross-linked acrylamide polymers were used to alleviate technical difficulties in cryosectioning and autoradiographic processing of marine arthropods (Pycnogonum litorale), larval insect tissues (Sarcophaga bullata), and amphibian (Xenopus laevis) oocytes. Rapidly polymerized cross-linked acrylamide was used to prepare sections from an animal with a hard thick cuticle, P. litorale. Fragmentation and compression artifacts caused by tissue density differences were avoided and internal tissues were well preserved. Our results indicate that closely applied external support eliminates difficulties typically associated with arthropod cryosectioning. Pre-polymerized, uncross-linked acrylamide provided similar protection when used for sectioning larval tissue from S. bullata. Soft tissues under the cuticle were undamaged in tracheal and epidermal preparations. This polymer was also an excellent embedding material for soft tissues such as salivary glands, quickly penetrating tissue cavities and eliminating air bubbles. Uncross-linked acrylamide was used as an embedding and culturing medium for oocytes from X. laevis. The polymers were non-toxic and allowed the preparation of thin frozen sections containing as many as 50 large oocytes per section.
Bronson
D D
DD
Department of Cell Biology and Anatomy, University of North Carolina, Chapel Hill.
Bidmon
H J
HJ
Tomaschko
K H
KH
Stumpf
W E
WE
D020106:Acrylamide; D000178:Acrylamides; D000818:Animals; D001345:Autoradiography; D016616:Cryoultramicrotomy / Q000379:methods*; D004175:Diptera / Q000033:anatomy & histology; D007814:Larva / Q000033:anatomy & histology; D009865:Oocytes / Q000166:cytology; D016610:Tissue Embedding; D014982:Xenopus laevis
D000178:Acrylamides; D020106:Acrylamide