NetNotes
literature or histology methods texts. I’ve always used and trained others to mount sections on the slide. If processing through a clearing agent don’t let the slide dry out, which traps bubbles within voids in the section. Apply the mounting medium along one edge of the coverslip (usually the edge furthest from me and parallel to the slide). Hold coverslip at an angle of 30–60o
using forefingers to the rear and thumbs on edge closest
to me. Bring the back edge of the coverslip with medium down to the slide, then slowly lower the coverslip down onto the slide, controlling with your thumbs. Allow the wave front of mounting medium to displace any air. Ten I flip the slide over onto a Kimwipe to blot excess mounting medium for a few minutes. Te weight of the slide presses out excess. Ten flip the slide back upright and allow to harden. Oſten a bubble can be “massaged” out by gently pressing on the coverslip towards the nearest edge. Glen MacDonald
glenmac@uw.edu
We advise our users to always put their sections on the coverslip,
not on the slide. Te aberrations start showing only a few microns from the coverslips with reasonably high NA objectives. Te bubbles come from how you apply the mounting medium, not from the thickness of the glass you use. I agree with the advice that others gave in this thread. Not using a squeeze bottle to apply the mounting medium helps. Use a glass rod dipped in the medium to transfer it. If this is not possible, one needs to carefully flip the squeeze bottle, wipe the first drop of medium out and apply the following drop of medium on the slide/coverslip without allowing air back in the bottle. Cleaning the coverslips with 1M HCl greatly helps sections to attach. See this video:
https://www.youtube.com/watch?v=j5_JsPPQTxo. To help collect sections (or to cytospin) on the coverslip, simply tape the coverslip on a slide with masking tape. Tis has the added advantages
You will still get bubbles if you put the sections on the coverslip. I put
the coverslip flat on a Kimwipe, then a dot of mountant on the coverslip (you really need to play with the right amount for your application), then wet the mountant with a drop of liquid (either xylene based or PBS, again depending on application), then flip the slide, catch the drop on the slide and let capillary action do the rest. You can push out small bubbles, but use a tool, not your fingers or mountant smeared gloves, I use a metal probe. I then wick up the excess with a Kimwipe on the edge of the slide. Important! Don’t keep smearing the moutant, just start again if it looks like a mess. Remember smears are the enemy of good imaging! Caroline Miller
caroline.miller@
unitybiotechnology.com
You are very welcome to use our videos. My experience is that it is
easy to convince our users to put their sample on the coverslip. When we train them, they must prepare samples for the training. We simply tell them to make 2 samples in parallel, one the way they normally do it and one on the coverslip. It takes no extra effort, so they do it. Ten we show them the difference with a high NA objective and let them decide what they want. One only sees the effect of not preparing the sample in an optimal way when one looks at the images side-by-side Sylvie Le Guyader
sylvie.le.guyader@
ki.se
that you can store the coverslips in a slide box as you would do with slides and that you can write notes on the slide. Tis video shows how one can collect paraffin sections using a coverslip: https://m.
youtube.com/watch?v=cDQFG-Pdm0o&feature=
youtu.be. Sylvie Le Guyader
sylvie.le.guyader@
ki.se
Scanning Electron Microscopy for the
Life Sciences Heide Schatten
University of Missouri, Columbia US$120.00: Hb: 978-0-521-19599-7: 312 pp
Recent developments in scanning electron microscopy (SEM) have resulted in a wealth of new applications for cell and molecular biology, as well as related biological disciplines. It is now possible to analyze macro molecular complexes within their three-dimensional cellular microenvironment in near native states at high resolution, and to identify specifi c molecu les and their structural and molecular interactions. New approach es include cryo-SEM applications and environmental SEM (ESEM), staining techniques and processing ap plications combining embedding and resin-extraction for imaging with high resolution SEM, and advances in immuno-labeling. With chapters written by experts, this guide gives an overview of SEM and sample processing for SEM, and highlights several advances in cell and molecular biology that greatly benefi ted from using conventional, cryo, immuno, and high-resolution SEM.
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The Press currently publishes the Microscopy and Microanalysis (MAM) journal in conjunction with the MSA, which reaches 4,000 microscopists and is affi liated with 12 international microscopy societies. The series would be a natural development from this journal, and will take a broad view of the discipline, covering topics from instrumentation to imaging, methodology and analysis across physical science, materials science, biology and medicine. Books commissioned for the series will range from advanced undergraduate textbooks through to research and practitioner oriented monographs for researchers. The series aims to produce a coherent source of material, encouraging the communication and exchange of ideas across these divergent fi elds, ensuring that the series appeals to a broad community in the physical and life sciences.
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www.cambridge.org/us 800.872.7423
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www.microscopy-today.com • 2020 September
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