Measuring Protein Expression and Cellular Fluorescence [Mean Gray Value vs Integrated Density]

[@richterdavidoliver7782]

Your Videos are excellent, I have done a bit of quantitative Image analysis, and I wish I had them back in the day. Many thanks for the effort you put into them.

[@johanna.m.dela-cruz]

It’s heartwarming to receive your positive feedback. Thanks for your support.

[@oliviapietrobon5225]

Hello from Argentina! I've been following you for a long time, and your videos are always great! Do you have any on immunohistochemistry analysis? I know you sometimes organize courses, and I’d love to learn more about them. Thanks!

[@johanna.m.dela-cruz]

Thank you Olivia (@oliviapietrobon5225) for your continued support. What type of measurements do you want to do on your IHC images?

[@oliviapietrobon5225]

@@johanna.m.dela-cruz Hello, I am analyzing images where I used immunohistochemistry to mark androgen receptors. Although it's not the same, when I saw your analysis in immunofluorescence, I noticed that the analysis is done for each ROI (Region of Interest) that you create. ¿Can I do the same in the case of immunohistochemistry? Another question, when you analyze each ROI, do you get a value for each one,To create the graph, ¿did you take an average of all those values? I hope my questions are clear. Thanks, Johanna!

[@johanna.m.dela-cruz]

@oliviapietrobon5225 As long as you arr able to segment the regions you want to measure, then, you can get measurements from each ROI. I am not sure what stain you used, but if you have DAB, quantifying intensity won’t be a sensible way to characterize antigen expression.

[@oliviapietrobon5225]

@@johanna.m.dela-cruz Hello, I use de tunel technique and caspasa 3 (for this i use DAB), now another question, I see that you're making a graph of the measurements; I'd like to know if you're calculating an average of the values from each ROI. My concern is that sometimes the values can be very different between each ROI, wouldn't that result in a high deviation?

[@johanna.m.dela-cruz]

@@oliviapietrobon5225 yes, it's an avaerage.