Our laboratory has recently added a vascular stain (STL - a conjugated lectin) in our immunohistochemistry protocols. We are not sure of the best way to quantify the stain. Does anyone have experience with this?
Note, the study is comparing age groups, thus the goal is to assess any age-associated differences in vasculature within the regions analyzed.
Thanks in advance!
There is a way to calculate fluorescence intensity in FIJI. This is the reference that I have used and had success with. https://theolb.readthedocs.io/en/latest/imaging/measuring-cell-fluorescence-using-imagej.html I hope it helps.
Hi Daniel,
I quantify vessel area after IHC stain without counter stain. Assessment of the CD31‐stained vessel area was performed by computer‐assisted image analysis. The areas of representative vascularization were chosen and five high‐power fields (HPFs) for each sample (n = 5) were captured with a digital camera. The images were imported into the automated image analysis package (NIS‐element software, Nikon) to calculate the stained vessel area per HPF.
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