We are upgrading our SMLM system and I’m interested in getting a filterset to allow us to do SMLM with NIR dyes, e.g. AF700, Cy5.5 etc. Obviously don’t want any bleedthrough from AF647 - which is our goto SMLM dye, since the aim would be to multiplex. Does anyone have any experience with good single or multiple bandpass filters for this?
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I have used the following from Chroma:
Emitter:
Supplier Item: zet405/488/561/647-656/75 Custom multi-band laser emission filter, 25mm diameter.
Dichroic:
zt405/488/561/647/752rpc Polychroic laser mirror, 26x38x2mm thick.
These are 5 band filters (405, 488, 561, 649, 750) and so are really expensive. You might talk to Chroma as they are usually willing to make you whatever you want for a similar price to what they typically stock.
Looks like you want something that is a little shorter in NIR wavelength. I’d guess this won’t be easy as A647 still has substantial emission past 700. You might have better luck with 647 excitation and ratiometric detection.
Hi Hazen sorry for the slow reply - we are currently upgrading all the super-res equipment in our facility and I got sidetracked. The emitter and Dichroic look lilke really good option for us thanks for the suggestion. I think our end users will not be too fussy. However they are very ambitious and are already discussing 3 colour STORM. I realistically think this will be ‘orange’ - perhaps Janeila Fluor 549, I can get AF555 to blink a bit if not, AF647 as our favourite and then something 750 ish. We already have people trying to get 3 colour DNA PAINT going who will be delighted to add a 4th fluor as well. Much appreciated.