Hi everyone, I stained some human tissue sections and it turns out that the cells tend to auto-fluoresce. I've used up all my antibodies, so I don't have a way to redo the experiments with a blocker for autofluorescence. Does any of you know a way to eliminate autofluorescence through image processing?
Hi,
There is a technique known as non-negative matrix factorization, which can be extensively used for auto-fluorescence removal. It is a complete image processing technique.
You may refer to Autofluorescence Removal by Non-Negative
Matrix Factorization
by Franco Woolfe, Michael Gerdes, Musodiq Bello, Xiaodong Tao, and Ali Can
-Cheers!
Hi Anuradha,
I found this thread discussing removal of autofluorescence: http://imagej.1557.x6.nabble.com/background-removal-in-fluorescence-td3704817.html
Some contributors share information that might be useful for addressing your problem.
Cheers, Christian
Anuradha,
If you still have the specimens and they still have their specific fluorescence, one thing you might try would be to float off the coverslips (very gently, and whether this will work depends on your mounting medium) and then apply Sudan Black to the sections to mask the non-specific autofluorescence, and then try taking some new images. A Sudan Black protocol is attached in case you want to take a hack at this.
Good luck,
Dave Sherry
- Badges
- Science topic
- Similar topics
- Engineering
- Electrical Engineering
- Signal Processing
- Image Processing