I'm trying to quantify lipid fluorescence of microalgal cell stained with Nile Red. The problem is I don't have spectrofluorometry to count the intensity of lipid fluorescence. Is it possible & valid if I use image analysis software (ex: imageJ, CellProfiler, etc) to quantify fluorescence using pixel intensity?
Hello, I think you can use a software for image analysis for quantification of neutral lipids using Nile Red. But first you have to calibrate the number of pixels/lipids concentration using a standar method (GC analysis for example). Not forget to take good photograph with the correct exitation/emission wavelenght in the microscope. Also you have to change the color of the photo to a black/white photo in order to obtain reliable results.
Just adding to the answer of Claudio, don't forget to also check unstained cells to see if you have background from chlorophyl fluorescence.
Probebly doable, but make sure you don't saturate your image! Alternatively visit the labs next door...
- Badges
- Science topic
- Similar topics
- Geoscience
- Asia
- India