I am aware that immunohistochemistry is a qualitative or semi-quantitative technique, but I performed several anti-CD163 immunohistochemistries in order to study the presence/absence of M2 macrophages in patients with several conditions. However, I have never quantified the result of immunohistochemistry before, so in the optical microscopy images that I took I see several brown marks (I used DAB for detection) but I am unable to determine if those marks are unspecific or if they are actually staining macrophages. Is there a way of correctly determining which of those DAB stains actually determine macrophages?
Thank you in advance.
First of all you have to optimize your immunohistochemistry protocol using proper controls, e.g. tonsil. When you know the expected reaction pattern, then do your experiment with on-slide controls so you can monitor the reaction from slide to slide. You can see some examples and suggestions to protocols here:
https://www.nordiqc.org/downloads/assessments/31_38.pdf
- Badges
- Science topic
- Similar topics
- Biological Science
- Molecular Cell Biology
- Culture Cells
- Cell Line