Microwave stabilization versus chemical fixation. A morphometric study in glycolmethacrylate- and paraffin-embedded tissue.
A morphological and morphometrical study was performed on testicular cells after microwave stabilization of the tissue while immersed in phosphate buffered saline (PBS), 0.9 NaCl or Tris-HCl. Fixation in Carnoy's fluid without irradiation was chosen as a control chemical fixation method. After microwave stabilization or chemical fixation, the testes were embedded in paraffin or in plastic (glycolmethacrylate). An excellent morphology, comparable to that after chemical fixation in Carnoy's fluid, was observed in the plastic sections of tissue irradiated in PBS or NaCl, even when the sections were subsequently treated with an aggressive reagent at high temperature, required for the Feulgen reaction. The nuclear area of the microwave-stabilized Sertoli cells was 37-46% smaller in haematoxylin-eosin stained, paraffin sections in comparison with that in the glycolmethacrylate sections. The microwave-stabilized, paraffin-embedded tissue was much more vulnerable to the hot HCl treatment of the Feulgen staining than the chemically fixed tissue, resulting in an additional 10-20% decrease in nuclear size. The latter finding is particularly important for quantitative microscopy, where the Feulgen staining method is often employed.
Department of Cell Biology, State University of Utrecht, Medical School, The Netherlands.
H J
HJ
Van de Kant
D G
DG
De Rooij
M E
ME
Boon
D000818:Animals; D002467:Cell Nucleus / Q000648:ultrastructure; D004396:Coloring Agents; D004801:Eosine Yellowish-(YS); D006416:Hematoxylin; D006651:Histocytochemistry / Q000379:methods*; D008297:Male; D008689:Methacrylates; D051379:Mice; D008815:Mice, Inbred Strains; D008872:Microwaves; D010232:Paraffin; D010503:Periodic Acid-Schiff Reaction; D012394:Rosaniline Dyes; D012708:Sertoli Cells / Q000187:drug effects / Q000648:ultrastructure; D013194:Staining and Labeling / Q000379:methods*; D013737:Testis / Q000033:anatomy & histology*
D004396:Coloring Agents; C007722:Feulgen stain; D008689:Methacrylates; D012394:Rosaniline Dyes; C005044:hydroxyethyl methacrylate; D010232:Paraffin; D004801:Eosine Yellowish-(YS); D006416:Hematoxylin