What is the reason behind fluorescence in 4% PFA fixed tissue?

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Alaa Frak Hussain

The most common fixatives are aldehyde cross-linkers such as 10% formalin or 4% paraformaldehyde (PFA). ... Heating and dehydrating samples (commonly done in tissue IF) can also increase autofluorescence, causing greater emission in the red spectrum

Rajsekhar Roy

Alaa Frak Hussain Yes, aldehyde cross linking is the culprit (I would say so) and we are trying to reduce the autofluorescenece by treating the tissue with NaBH4. Could you please put an insight whether there is any problem we could face further if we use NaBH4 in rat skin tissue cryosection before performing IHC?

also to mention,

we will be working with brain tissue sections next... in that case could you please suggest whether to stick to the borohydride or, 10 mM CuSO4 in 50mM NH4Cl is better? As borohydride just work fine only in case of aldehyde fixation and and sudan back works good for endogenous fluoroscence of the tissue (lipofuscin).