Ultrastructural preservation of rat embryonic dental tissues after rapid fixation and dehydration under microwave irradiation.
Adequate preservation of the cells and matrix of mineralising tissues remains difficult, as organic components and initial mineral deposits may be lost during conventional processing for electron microscopy. In this study, we have reduced significantly the processing time using microwave irradiation. Rat molar tooth germs were fixed in 4% glutaraldehyde + 4% formaldehyde with 0.1 M sodium cacodylate in a laboratory microwave oven for two periods of 20 s with a maximal temperature of 37 degrees C. After conventional washing and post-fixation, specimens were dehydrated in graded ethanols under microwave irradiation for a total of 7 min 20 s. For comparison, some specimens were processed by conventional methods. After embedding, ultrathin sections were examined by electron microscopy. In differentiating ameloblasts and odontoblasts, plasma membranes, mitochondria, rough endoplasmic reticulum, the Golgi complex, together with all other cytoplasmic organelles exhibited excellent preservation. Microtubules, microfilaments and coated vesicles were particularly evident. Crystal-like mineral deposits were conspicuously present in relation to dentine matrix vesicles and collagen fibrils as well as in enamel matrix. The matrix of forming enamel had a globular electron-lucent appearance. It is concluded that this is a rapid method which provides a preserved or even improved morphology.
Massa
L F
LF
Department of Histology and Embryology, Institute of Biomedical Sciences, University of São Paulo, SP, Brazil.
Arana-Chavez
V E
VE
D000565:Ameloblasts / Q000648:ultrastructure; D000818:Animals; D003593:Cytoplasm / Q000648:ultrastructure; D008872:Microwaves; D009804:Odontoblasts / Q000648:ultrastructure; D051381:Rats; D016707:Tissue Fixation / Q000295:instrumentation / Q000379:methods*; D014083:Tooth Germ / Q000196:embryology* / Q000648:ultrastructure*