How to detect flavonoids in brain tissues via fluorescence microscopy? It is reported that 2-Aminoethyl diphenylborinate (ADPB) can interact with flavonoids and enhance the fluorescence intensity of flavonoids in a cell which can be imaged via fluorescence microscopy (see the ref. BioTechniques 66: 65-71, 2019. DOI: 10.2144/btn-2018-0084). Can this technique be followed during tissue imaging (in vivo test) in rat? Thank you in anticipation.
There is a huge difference in uptake of molecules and detection between a simple organism like Dictyostelium and to that of rats.
You have to answer the following for starters:
- You are mentioning in vivo - do you have the type of setup (microscope and imaging setup like multiphoton microscope, etc.)?
- Does the molecule pass across the BBB?
- What is the half life of the molecule in vivo?
- Toxicity?
- How it is metabolized?
- Mode of administration
- Dosage
There are so many questions that has to be asked even before you think of in vivo imaging. Good luck.
I am assuming you are planning to use primary cell line with a fluorescence microscopy imaging. Simple flavonoids most often excitable with 405 nm laser line in a fluorescence microscope. Typically emits in the blue-green region. However, based on the typical absorption coefficients and fluorescence quantum yields for flavonoid based compounds, your detection limit must not be below typically 200 nM or so. Otherwise, they ,may be challenging to visualize.
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