A simple method for processing individual oocytes and embryos for electron microscopy

Abstract

Summary A simple method for handling individual specimens that must be processed either for scanning or transmission electron microscopy studies is described. For scanning microscope processing, dehydration is carried out with samples enclosed in small cages made from TAAB capsules in which top and bottom are substituted by plankton nets, and for transmission electron microscopy, samples are pre‐embedded in agarose. This procedure significantly reduces mouth pipetting, dissecting microscope observations, is less labour intensive and, most importantly, reduces sample loss.

Characterization of an oil-degrading Microcoleus consortium by means of confocal scanning microscopy, scanning electron microscopy and transmission electron microscopy

“…They were then centrifuged and dehydrated in successively increasing concentrations of acetone (30, 50, 70, and 100%). Subsequently, the samples were processed according to the method described by Nogués et al (1994) (this method describes the use of specific capsules in which the sample is placed and then dehydrated without loss or alteration) and dried by critical-point drying. Finally, all samples were mounted on metal stubs, coated (thus avoiding charging the samples and ensuring good image quality) with a 96 nm layer of gold (this ensures better secondary-electron emission and, therefore, better image quality).…”

New method for the treatment of sperm samples for ultrastructural study based on the use of animal tissues as biological containers

“…To study the ultrastructure of free or in-suspension cells by means of TEM it is necessary either to centrifuge the samples previously to the fixation, in order to obtain a cellular pellet (Gonzalez Santander, 1970;Krogenaes et al, 1994;Quinn et al, 1969) or to immerse the samples in viscous gels. Among the latter, the most used methods are those in which the samples are preembedded in agarose (Nogues et al, 1994;Watanabe et al, 1988) or those that use the agar encapsulating technique (Pihakaski et al, 1990).…”