A question for fluorescence microscopists

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jmc
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A question for fluorescence microscopists

Post by jmc »

I was wondering, do those of you doing fluorescence microscopy use quartz or fused silica microscope slides and coverslips, or do you just use normal glass ones?
Jonathan Crowther

Pau
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Re: A question for fluorecence microscopists

Post by Pau »

You only need quartz slides and covers if you use short UV radiation.
Fluorescence excitation microscopy is mostly done with blue, violet and long UV (360nm to 400nm) light and most glass slides and covers are adequate (some ones can show excessive autofluorescence, particularly oldest and bad quality ones).
Pau

jmc
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Re: A question for fluorescence microscopists

Post by jmc »

Ok thanks Pau. Yes, I was wondering whether autofluorescence would be a problem even with longer wavelength UV. Are then any hints for buying glass slides with low fluorescence?
Jonathan Crowther

Ichthyophthirius
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Re: A question for fluorescence microscopists

Post by Ichthyophthirius »

Hi,

In slides, autofluorescence is associated with iron content. The best slides have low iron content and are marginally more expensive because they rely on low iron content raw materials.

I would recommend just going for quality slides - from Menzel or Marienfeld. They also make cover glasses from low autofluorescence Schott D 263 M. It's a shame they are a bit expensive for end users, because when bought in bulk they are very affordable.

Cover glasses are probably more important than the slides as they are closer to the object and the high intensity illumination cone.

jmc
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Re: A question for fluorescence microscopists

Post by jmc »

That's good to know Ichthy. My glass ones are basic cheap ones, and that probably explains why they light up under UV.

I see Thorlabs do the D263M coverslips, and in bulk quantities if needed.
Jonathan Crowther

jmc
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Re: A question for fluorescence microscopists

Post by jmc »

Posting this in case it is of interest to anyone. I've done a little bit of work on glass and quartz / UV fused silica slides and coverslips, looking at UV transmission and fluorescence with different UV wavelengths.

https://jmcscientificconsulting.com/uv- ... overslips/

Mods - I hope this is Ok to post, please let me know if not.
Jonathan Crowther

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Re: A question for fluorescence microscopists

Post by rjlittlefield »

jmc wrote:
Sun Aug 23, 2020 4:30 am
Mods - I hope this is Ok to post, please let me know if not.
That's a great article -- thanks for the link, and the work behind it!

--Rik

jmc
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Re: A question for fluorescence microscopists

Post by jmc »

No problem Rik, glad you liked it. The idea with those updates is to share as much as I can of my experiences while I learn about making a UV microscope.
Jonathan Crowther

Lou Jost
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Re: A question for fluorescence microscopists

Post by Lou Jost »

Excellent, this is great information, thanks! I had seen blanket recommendations that a normal thin glass coverslip does not block UV light, but your thoroughness reveals that this (and fluorescence too) changes dramatically with just a small change in excitation wavelength. Very nice thougthtful demonstration.

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Re: A question for fluorescence microscopists

Post by Pau »

Very interesting work, thanks for sharing it!
Pau

jmc
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Re: A question for fluorescence microscopists

Post by jmc »

You're welcome, happy to share results like this as and when I have them.
Jonathan Crowther

Ichthyophthirius
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Re: A question for fluorescence microscopists

Post by Ichthyophthirius »

Nice post! The transmission results line up well with the published data https://www.schott.com/en-gb/products/d ... al-details

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