I am planning to make a buffer solution to make cy5 dye photoswitchable. The buffer will contain:

-TN buffer (50 mM Tris (pH 8.0) and 10 mM NaCl)

-0.5 mg/ml glucose oxidase

-40 µg/ml catalase

-10% (w/v) glucose

-143 mM βME

as done in Dempsey, G. T. et al., (2011). Evaluation of fluorophores for optimal performance in localization-based super-resolution imaging, 8(12).

I also want to bind the cy5 chemical to my glass and gold substrate, so I planned on using the βME as a thiol linker to gold (also unsure if this is appropriate) and the 1,5-Diamino-2-methylpentane as an amine linker to cy5 nhs ester. Finally I was going to use SMCC to link the βME to the 1,5-Diamino-2-methylpentane. (the cy5 that I hope to link with these will already be in the buffer solution with βME described above).

I am a physicist and not a chemist, so I am not confident about the safety of mixing all of these chemicals or how well this method will work - any insight is helpful.