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A SYNCHROTRON X-RAY STUDY OF THE CHANGES OCCURRING IN THE CORNEAL STROMA DURING PROCESSING FOR ELECTRON-MICROSCOPY

FULLWOOD, N J and MEEK, K M (1993) A SYNCHROTRON X-RAY STUDY OF THE CHANGES OCCURRING IN THE CORNEAL STROMA DURING PROCESSING FOR ELECTRON-MICROSCOPY. Journal of Microscopy, 169. pp. 53-60. ISSN 0022-2720

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Abstract

Using a high-intensity synchrotron X-ray source, the structural changes occurring in the corneal stroma were monitored during each stage of several different processing runs for the transmission electron microscope (TEM) and scanning electron microscope (SEM). The parameters studied were interfibrillar spacing, intermolecular spacing, D-periodicity and fibril diameter. The processing schedule that produced the least changes in spacings for TEM specimens involved extended fixation in glutaraldehyde followed by low-temperature embedding in Lowicryl K4M resin. However, interfibrillar material was better preserved after embedding in LR White resin or Nanoplast. Almost every processing stage for electron microscopy produced significant changes in one or more structural parameters in the cornea. Glutaraldehyde fixation significantly increased the intermolecular spacings, while resin infiltration and resin polymerization each resulted in shrinkage of all the spacings monitored. Critical-point drying for SEM specimens resulted in considerable shrinkage in all three spacings, but was still preferable to air drying, which caused reduction in the order of the fibril packing, resulting in loss of the interfibrillar X-ray pattern. Perhaps the most drastic effect was caused by post-fixation in osmium tetroxide, which resulted in loss of the intermolecular pattern, and also increased the amount of shrinkage in the interfibrillar spacings and the D-periodicity which occurred during later stages of processing.

Item Type: Article
Journal or Publication Title: Journal of Microscopy
Uncontrolled Keywords: CORNEA ; X-RAY SYNCHROTRON ; ELECTRON MICROSCOPY ; PREPARATION ; RESINS ; COLLAGEN FIBRILS
Subjects: UNSPECIFIED
Departments: Faculty of Health and Medicine > Biomedical & Life Sciences
ID Code: 49175
Deposited By: ep_importer_pure
Deposited On: 28 Jul 2011 15:11
Refereed?: Yes
Published?: Published
Last Modified: 26 Jul 2012 19:25
Identification Number:
URI: http://eprints.lancs.ac.uk/id/eprint/49175

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