Hi everyone, l have a question.
I made cell cultivation on a biomaterial containing chitosan and gelatin, fixed my cell culture samples on the 14th day with 4% paraformaldehyde and performed H&E staining. However, while the scaffold has cross-sectional images, the cell is not in the environment. why could this be caused?
Note: there is a picture attached.
Thanks.
I also tried several times and tried each time by changing one of the factors such as fixative, dye (H&E), xylene, but again I could not see any cells.
Ayfer Koyuncu Have you tried visualizing the samples before fixation? If the cells are visible then, change the harshness of fixation or washing procedure.
Nabodita Sinha thanks for your opinion. Since I detected the presence of cells on the biomaterial in MTT assay and SEM analysis, it was very interesting that the cells were not visible in the H&E staining of the same sample. When I made a literature review, it was suggested that it should be kept in the fixation solution for at least 4 hours, but I kept it for 1 hour. Therefore, I think that the cells do not hold in this period and leave the environment in washing with dPBS. I will try the fixation time again for a minimum of 4 hours and a maximum of 1 day.
I hope this is the problem.
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