Usually to visualise proteins or biological polymers in Transmission Electron Microscopy, negative staining with uranyl acetate or phosphotungstic acid is done but i simply drop casted my protein-polymer (alginate) complex on TEM grid and my proteins are seen as dark dots even without negative staining, the buffers used were simple sodium acetate and sodium phosphate.
Dear Mayur,
That seems like a curious case. I was wondering how your procedure is. Which steps have you done exactly?
Did you also perform a negative control somehow? Like only buffers w/o protein complex?
Dear Mayur,
You did not reveal much detail about your protein of interest - could it be bound with some metals? If anything appears dark in TEM, it has something which prevents electron transmission, usually heavy metals nuclei. Some cells/structures, such as erythrocytes, contain enough metals to be visible in TEM, even without additional stains.
@michal kulus actually my proteins belong to lysozyme family which tend to crystallise and i got the answer in some papers that crystallites are formed which appear dark, thank you for taking time🙂
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