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Updated: Jul 23, 2026
Serial Block-Face Scanning Electron Microscopy (SBF-SEM) of Biological Tissue Samples
Published on: March 26, 2021
Comparison of conventional and microwave-assisted processing of mouse retinas for transmission electron microscopy
K D Wendt1, C A Jensen, R Tindall
1University of Missouri School of Medicine, Mason Eye Institute, Columbia, MO 65212, USA.
Abstract:
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.
Insights
Microwave-assisted fixation significantly reduces artifacts in mammalian retinal tissue for transmission electron microscopy (TEM). This faster method improves ultrastructural preservation, especially in photoreceptor cells, saving valuable preparation time.
Area of Science:
- Ophthalmology
- Microscopy
- Cell Biology
Background:
- Conventional transmission electron microscopy (TEM) preparation of mammalian retinal tissue is time-consuming.
- Standard methods often introduce ultrastructural artifacts, particularly in the photoreceptor cell layer, such as gaps between disc membranes and cells.
Purpose of the Study:
- To evaluate the efficacy of a rapid microwave-assisted fixation and processing protocol.
- To determine if microwave irradiation improves ultrastructural preservation in retinal tissues, specifically reducing photoreceptor artifacts.
Main Methods:
- Mammalian retinal tissues were prepared using both conventional fixation/processing and a rapid microwave-assisted protocol.
- Ultrastructural preservation quality was assessed, with a focus on photoreceptor cell layer artifacts.
Main Results:
- Overall retinal ultrastructural preservation was comparable between conventional and microwave-assisted methods.
- Microwave irradiation significantly reduced the magnitude of photoreceptor artifacts compared to conventional techniques.
Conclusions:
- Microwave-assisted specimen preparation for TEM offers superior ultrastructural preservation of retinal tissues.
- This method substantially decreases preparation time while improving the quality of electron microscopic images.
