Fluorescence Filters: Choosing a Filter Set

Can you confidently choose an optimal filter set to image your fluorophore? This video walks you through the why and how of picking filters that will allow you to collect the most light from your fluorophore, while reducing background and increasing contrast. Enjoy!
Visit FPbase.org/microscopes to generate a custom spectraviewer for your microscope.
French subtitles were provided by BioImaging North America.

[@t.b.gandek4257]

This would have been so useful when I started my PhD. This was the explanation I was looking for.

[@alexandrevaliquette3883]

You did not know that when you started your PhD? Where did you study before?

[@dmarting6014]

Wow. Thank you so much for this. I work for a company that is just getting into work in epifluorescence. I am certainly a bit over my head and this was a massive help.

[@autismunion]

you are great example to ''how to teach''!!

[@lizs7827]

This was wonderful, very clearly explained. Thank you so much!!!

[@Walaa918]

Great explanation. Thank you very much.

[@mariociencia12]

Amazing! Fantastic! Astounding! Incredible! Mighty! Much better than Marvel's movies!

[@RadhikaSharma-oi4ne]

Wow. I am a phd student learning fluorescence microscopy at the moment for some of my experiments. This totally blew my mind away and cleared most of my misconcepts. Thank you!

[@Microcourses]

Yay!

[@alexandrevaliquette3883]

Whaaaat? In 2024, how a PhD student in microscopy not know this very basic stuff?

[@yahelabraham9821]

excellent video

[@flyingpinguu]

Super helpful!

[@niceday2015]

Thank you very much. Great teacher

[@Microcourses]

You are welcome!

[@nandubattula2371]

so much helpful.......

[@ladushky1]

Thank you so much for the video! Could we choose the specific excitation and emission filters in widefield microscopy as well? I guess this optimization is only possible in confocal microscopy since in widefield the filters are all in one unit, is that correct?

[@Microcourses]

Yes, you can! Widefield filters are mounted into a single holder (often called the filter cube), but they use the same set of filters described here.

[@bilz0r]

At 4:24, I think you got the color of your labels mixed up.

[@Microcourses]

I did! Thanks for pointing that out. Hopefully it won't be too confusing for anyone.

[@emirselman3984]

Is it the beam splitter or the emission filter that blocks out all the millions of extra exication photons that overdrown the emitted photons?
In the beginning you said the emission filter was to remove all this overdrowning blue light, but in the end you make it seem like the emission filter is only for the stray light, i.e. the few photons that transmit the beam splitter from the excitation filter

[@Microcourses]

The beam splitter blocks some, the emission filter blocks the rest. How much each contributes depends on the transmission properties of each filter in the set.