I subcutaneously performed implantation in rats and harvested the sample after 1, 2, 3, and 6 months. The implant is decellularized human skin. The procedure of how my lab did it was drawn in the attached picture.
Then I carried out IF staining with alpha-SMA, and Vimentin for the sample to monitor fibroblast migration into the implant. The magnification of the picture was 20X, therefore, the region that appeared in the picture is the implant area. I observed cell morphology and also something that looks like a "thread"(?), a very thin structure and don't look like cells at all. I'm not sure what that is and I tried to find references but I could not find anyone who did the same staining for human skin implant.
Could someone please help me with this problem? Please let me know if you have any opinion on this. Thank you so much
Hi Xin Rui Zhang - it is difficult to guess. It looks rather like fibrous elements or residual of blood vessels. My main concern is that the DAPI staining does not look correct - the shape of the nuclei (as DAPI stains DNA mostly) is abnormal. Did you use confocal microscope? I suspect that the channels for different stainings were not well separated or there also could be an overexposure in the DAPI channel. Possibly, the concentration of the residual DNA in the decellularized skin implants was quite high, and you are seeing the diffuse DAPI staining of the fibers of the decellulrized tissue.
Anna Guller Hi, thank you so much for your response. Actually, in the sample I showed here, there were supposed to have no cells. We tried to evaluate the fibroblast cell invasion into the ECM implant after 1, 2, 3, and 6 months and the picture was after 1 month. I checked another month's slide, they did have cell infiltration into our implant and the nuclei exhibited the common shape as usual. Therefore, I question what are those fibrous elements in the 1 month sample... If you may come up with any ideas, please let me know...
Anna Guller oh sorry i forgot to mention that we used confocal microscope and it's vimentin staining!
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