Is it cryo method or normal conventional method ? Please give a standard protocol for the analysis with Citation.
Nagababu Konagala
I do not have any expertise in this area. But I am very much familiar with TEM. So I've given my point of procedure and provided some sources which may help you. Chemical Fixation Dehydration Embedding Sectioning Ultra-thin sectioning (60-100 nm thickness) using diamond knives Staining Double staining with: Uranyl acetate (2 minutes) Lead citrate (1 minute) Enhances electron contrast. Imaging Analysis at 80-120 kV accelerating voltage in TEM
Conventional TEM Sample Preparation Protocol
- Michielsen K, Stavenga DG. Gyroid cuticular structures in butterfly wing scales: biological photonic crystals. J R Soc Interface. 2008 Jan 6;5(18):85-94. doi: 10.1098/rsif.2007.1065. PMID: 17567555; PMCID: PMC2709202.
- https://pure.rug.nl/ws/portalfiles/portal/14499057/c2.pdf
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Irene Wacker
No, cryo would make no sense, because butterfly wings do not contain water. They can be viewed as a material (consisting mostly of chitin) because they are totally dry. Therefore you do not need a chemical fixation either. You can use a very simple embedding protocol (Siddique et al, Scientific Reports | 6:36204 | DOI: 10.1038/srep36204) with Epoxide resin before cutting ultra thin sections for TEM. In our paper mentioned above we imaged the sections in an SEM, but of course a TEM can also be used when the sections are thin enough, meaning about 70nm. Hope that helps!
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