Reflection contrast microscopy for high resolution detection of (3)H-estradiol in ultrathin sections of human stratum corneum |
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Authors: | Neelissen J A Schrijvers A H Junginger H E Boddé H E |
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Affiliation: | AstraZeneca R&D S?dert?lje, Pharmacokinetcs and Biopharmaceutics, 151 85 S?dert?lje, Sweden. jan.neelissen@astrazeneak.com |
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Abstract: | A single autoradiographical method for light and electron microscopy (LM and EM) is presented. Human skin, containing (3)H-estradiol ((3)H-E2) after an in vitro permeation experiment, was processed via a non-extractive tissue preparation protocol, comprising cryo-fixation, freeze-drying, osmium tetroxide vapor fixation, and Spurr resin embedding. Semithin sections were processed for LM autoradiography, while ultrathin sections were processed both for high-resolution LM and EM autoradiography. The autoradiographs were visualized by bright-field microscopy (BFM), reflection contrast microscopy (RCM), and transmission electron microscopy to evaluate the potentials of RCM visualization in high-resolution LM autoradiography. RCM visualization of ultrathin vs. semithin resin sections showed an improved stratum corneum morphology. Histological staining was superfluous. The localization of (3)H-E2 in human stratum corneum using high-resolution LM autoradiography and RCM was as accurate as with high-resolution EM autoradiography. |
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