I have done one time TEM experiment to observe the peanut protein(Ara h1).The process was simply with 3 microlitre protein sample diluted into 1 ml PBS(Buffer) and then a drop of diluted sample was applied to the grid and observed under TEM machine. Unfortunate nothing was seen.
what was the exact problem for TEM in this case?
Once any researchers have methodology about sample preparation for TEM, especially protein, requested to provide.
Abul, honestly:
have you tried to do any literature seaech for your problem? TEM specimen preparation technique (esp. "negative staining methods") do[es] not / did not start today.....
("was applied to the grid"....in utmost consequence you failed to find your protein due to the holes / grid mesh)
But I have tried to find negative staining method which is related to the protein but I did not find anything related to the protein. If you have any
thing regarding this, Kindly give me the link or my email.
Abul,
here you have same classics:
M. A. Hayat, Sara E. Miller: Negative Staining McGraw-Hill Publishing Company, 1990
https://books.google.cz/books/about/Negative_Staining.html?id=0h5rAAAAMAAJ&redir_esc=y
Harris, J. R. Transmission electron microscopy in molecular structural biology: A historical survey. Archives of biochemistry and biophysics, Elsevier BV, 2015, 581, 3-18
http://dx.doi.org/10.1016/j.abb.2014.11.011
Article (Ohi2004) Ohi, M.; Li, Y.; Cheng, Y. & Walz, T. Negative Staining and Image Classification - Powerful Tools in Modern Electron Microscopy. Biological procedures online, Springer Nature, 2004, 6, 23-34
http://dx.doi.org/10.1251/bpo70
and many more
Abul,
I can send you a mail with the protocol to negatuve stainning of protein.
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