Imaging nervous system tissue with TEM. I've come across lots of examples of various artifacts related to fixation and contamination, but I would love to see some examples that have been over-stained with the various traditional counterstains: osmium, uranyl-acetate and lead citrate. Is anyone aware of a good publication with such images or perhaps would care to share their own?
I just want to remind you that staining today is not the same as it was 10-15 years ago. Nowadays almost everybody uses digital cameras, which greatly improve contrast of images. So, much less (if any) post staining is needed. For example, when working with immunogold, I rarely use any other stain but OsO4. In this way gold particles are much better visible. For me overstaining when working with immunogold is when some parts of image are as dark as gold particles. Each task, each method of image acquisition has its own definition of overstaining.
Precipitation is a different story, it is still here. May be this site will be of some use to you:
https://www.leica-microsystems.com/science-lab/brief-introduction-to-contrasting-for-em-sample-preparation/
Hi Jonathan,
If you do not restrict the images to nervous system tissue, a good source would be Biomedical Electron Microscopy by Maunsbach & Afzelius ( https://doi.org/10.1016/B978-0-12-480610-8.X5000-8 )
especially chapters 2 (fixation) and 9 (section staining).
The original printed version of the book has very nice images, the PDF version is also adequate. I wouldn't recommend buying the re-print of this book (it was apparently made by scanning an original copy of the book and then printing it again, the image quality is horrid)
best,
Rob
I just want to remind you that staining today is not the same as it was 10-15 years ago. Nowadays almost everybody uses digital cameras, which greatly improve contrast of images. So, much less (if any) post staining is needed. For example, when working with immunogold, I rarely use any other stain but OsO4. In this way gold particles are much better visible. For me overstaining when working with immunogold is when some parts of image are as dark as gold particles. Each task, each method of image acquisition has its own definition of overstaining.
Precipitation is a different story, it is still here. May be this site will be of some use to you:
https://www.leica-microsystems.com/science-lab/brief-introduction-to-contrasting-for-em-sample-preparation/
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