I currently work with fluorescence microscope to observe generative and vegetative cells within hibiscus rosa sinensis pollen. I used DAPI to stain pollens but there is no light that indicate the existence of both generative and vegetative cells. I simply stained the pollens with 50 ppb DAPI without other procedure. I have referred this steps from Sexual Plant Reproduction textbook from Cresti and Tiezzi, here is the link : le.ac.uk/bl/phh4/openpubs/SPR_HH_pollen_stigma_crestiLR.pdf. From the book, it simply states "For DAPI staining, fresh or fixed pollen is stained on a slide in a drop of 0.005% DAPI (diamidinophenylindole), and the nuclei observed under a fluorescent microscope after 5-30 min (UV excitation, blue DAPI fluorescence).
The result is make me confused because there was no fluorescent light but only reflected light by pollen itself.
I will be very glad if anyone can suggests me what should i do or highlights an error in my steps.