FluoroGold has a similar structure to the very common lipophilic antero/retrograde fluorescent tracer DiI-dyes (if you disregard the alkene-chains). They come is different colours and tracing speeds.
We tested some fixable variants, SP-DiI and CM-DiI, that are not washed out during CLARITY.
CM-DiI in short incorporates a mildly thiol–reactive chloromethylbenzamido substituent that confers aldehyde fixability via conjugation to thiol-containing proteins. SP-DiI has two sulfophenyl groups that confers fixability. FM 1–43FX, which is less similar to DiI, is a lipophilic styryl dye with an aliphatic amine side chain for aldehyde fixability.
They are all retained in the tissue and strongly fluorescent after clearing with CLARITY.
FluoroGold has four amine ends, which makes it very aldehyde fixable, and CLARITY-compatible. I do not expect you need to change you your FluoroGold staining protocol.
https://www.nature.com/articles/srep32674
Hello,
The only issue with using FG for whole-mount brain imaging is that the UV excitation necessary to image FG has a relatively small penetration in tissue, even after clearing.
If it is a possibility, I would recommend using a anti-FG antibody for immunostaining. This way you can use a secondary antibody conjugated to a far-red dye (such as Cy5 or Alexa-647), which will greatly improve your imaging. I have tested the FG antibody from Protos Biotech in the past and it worked quite well: http://www.protosantibody.com/
You may also be interested in looking into the iDISCO clearing protocol, which is optimized for immunostainings and is much faster than CLARITY: https://idisco.info/
I hope this helps!
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