Wendt K D, Jensen C A, Tindall R, Katz M L
University of Missouri School of Medicine, Mason Eye Institute, Columbia, MO 65212, USA.
J Microsc. 2004 Apr;214(Pt 1):80-8. doi: 10.1111/j.0022-2720.2004.01310.x.
Conventional fixation and processing of mammalian retinal tissues for transmission electron microscopic (TEM) examination is slow and produces ultrastructural artefacts in the photoreceptor cell layer. Among these artefacts are gaps between photoreceptor outer segment disc membranes and between photoreceptor cells in the region of the retina where the cell nuclei are located. A study was undertaken to determine whether a much more rapid microwave-assisted fixation and processing protocol would have an effect on the quality of ultrastructural preservation of the retina, particularly on the photoreceptor cell artefacts. The overall ultrastructural preservation of the retina was similar for the conventional and microwave-assisted techniques. However, the magnitudes of the photoreceptor artefacts were significantly reduced when microwave irradiation was used during primary fixation and processing. It is clear that, at least for the retina, employing microwave irradiation during specimen preparation for TEM results in superior ultrastructural preservation with a substantial reduction in the time required for sample preparation.
用于透射电子显微镜(TEM)检查的哺乳动物视网膜组织的传统固定和处理方法耗时较长,并且在光感受器细胞层会产生超微结构假象。这些假象包括光感受器外段盘膜之间以及视网膜细胞核所在区域的光感受器细胞之间的间隙。开展了一项研究,以确定一种更为快速的微波辅助固定和处理方案是否会对视网膜超微结构保存的质量产生影响,尤其是对光感受器细胞假象的影响。传统技术和微波辅助技术对视网膜的整体超微结构保存效果相似。然而,在初次固定和处理过程中使用微波辐射时,光感受器假象的程度显著降低。显然,至少对于视网膜而言,在TEM标本制备过程中采用微波辐射可实现更好的超微结构保存,同时大幅减少样品制备所需的时间。