which LED and filter and mirror of the miniscope to works with RCaMp

[Chenglin]

To works with RCaMp and the excitation and emission of this protein are at 587 nm and 610 nm. could you help to tell me which LED and filter and mirror of the miniscope should I select?
To work with the GCaMP6, I have ordered the ET470/40x, T495lpxr and ET525/50m from the CHROMA company. But not sure about the pieces work with RCaMp. Hope you could share some ideas. Thank you very much!

[Daniel Aharoni]

Hi Chenglin,
Great question. You can use Chroma's website, https://www.chroma.com/products/fluorochrome, to find filter sets specifically designed for whichever fluorophore you want to image. You want to pick a filter set that uses 1mm thick glass and you can ask Chroma to dice the filters to the size required by the Miniscope. We have had good success imaging jRCaMP1a using ET545/25x, T565lpxr, ET605/70m (Chroma set #49004).

As for the LED, the easiest option is to pick a different color variant in the LUXEON REBEL product line, http://www.digikey.com/products/en/optoelectronics/led-lighting-color/125?k=&pkeyword=&s=38296&s=40968&FV=ffe0007d%2Cfffc0588&mnonly=0&newproducts=0&ColumnSort=0&page=1&quantity=0&ptm=0&fid=0&pageSize=25. You just need to pick an LED with a peak wavelength near the middle of the excitation filter band.

[Chenglin]

Thank you very much!

I will try with the filters and mirrors you suggested.

[Chenglin]

Hi,

I think I will try with the 2 LED below, do you think if they are OK?

LED Lighting Color LUXEON Rebel Color Green 530nm (Typ) 2-SMD, No Lead, Exposed Pad   digikey   1416-1036-1-ND   25   72 USD   http://www.digikey.com/product-detail/en/lumileds/LXML-PM01-0100/1416-1036-1-ND/3961141

LED Lighting Color LUXEON Rebel Color Amber 590nm (Typ) 2-SMD, No Lead, Exposed Pad   digikey   1416-1033-1-ND   25   52 USD   http://www.digikey.com/product-detail/en/lumileds/LXML-PL01-0050/1416-1033-1-ND/3961138

[Daniel Aharoni]

Yes. The first one will be a good match.

[Chenglin]

Thank you for your great help!

[jwghim]

Hi Daniel,
I'm trying to image jRGECO1a, but haven't succeed to get any images of red indicators.
I have using filter set ET560/40X, T585lpxr, ET620/60m and Luxeon green led LXML-PM01-0100.
Actually that is the filter set for mCherry. I haven't get images of mCherry with it, either.
Could you take a look at my setup and give me some suggestions?
Thank you in advance!

[Daniel Aharoni]

Hi jwghim,
Have you been able to image jRGECO1a or mCherry with a different microscope? Are you sure it is expressing and that your lens implant is at the proper depth. It sounds like your LED and filter set should be fine to image these fluorophores.