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This is a more generalized question of my other question here.

I want to know if it is possible to determine a chemical compound in a non-homogeneous sample. I am asking this because we are aiming to create a device that will detect histamine level in fish without homogenizing the samples. In all the journals we've read (which are a lot), fish samples are homogenized (blended + chemically-treated) before histamine is determined.

I am a computer engineering student with basic knowledge on biochemistry. I'm researching on this because this is our thesis topic (which our adviser has forced recommended to us).

Any help/recommendation regarding this is appreciated. Thank you!

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You should not actually homogenize samples because histamine works locally. If you know which region is releasing histamine then you can just study those regions. You can actually create a transgenic fish with GFP that is a target of histamine receptor signaling.

Using mathematical models or statistical correlations you can obtain the level of histamine based on fluorescence.

Activated histamine receptor H1 causes production of inositol triphosphate (IP3) which in turn causes release of calcium form ER. You can use Cameleon-GFP to sense the change in calcium levels. However, there are many events that may result in calcium release; you should carefully design your experiment.

This is another possibility (just a guess)- a FRET based study using GFP-YFP FRET Pairs (or other variants). Since histamine receptors are G-protein coupled receptors you may fuse the GFP to alpha subunit and YFP to beta subunit. When upon signaling, the subunits dissociate, you would see a decrease in FRET.

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It is correct that imaging-based studies are super-awesome, but question seems to be about chemical. I don't know if there are antibodies available for histamine, if yes, then you can fix and label all sites of histamine.

But imaging through fish tissues (say, more than 1 mm) is close to impossible with molecular resolution and sensitivity.

My best guess would be following. If you are OK with blood level measurements, then try spectroscopic approach. That is measure Raman NIR scattering or fluorescence/absorption in UV-visible regions of light spectrum.

Fun option is to develop chemical-reaction based assay. Say, engineer reaction that will modify histamine using radioactive-labeled compound and then measure how fast/slow amount of the compound decreases

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