Autofluorescence images

Pau · Post by **Pau** » Mon Dec 12, 2016 11:04 am

Still testing my setup for autofluorescence

Polypodium cambricum immature sorus, stacked images

1. 4X Pl Apo, 6.4X on APSC sensor. Blue 445nm LED, LP emission filter

2. UV 365nm LED 10X Pl Fluotar, LP emission filter (2.5s exposure at ISO 100)

Charles Krebs · Post by **Charles Krebs** » Mon Dec 12, 2016 11:11 am

Looking good!
Excellent images!

discomorphella · Post by **discomorphella** » Mon Dec 12, 2016 12:20 pm

Looking good indeed Pau. Now just get yourself some Congo Red, Calcofluor White and Acridine Orange (and GLOVES) and you're set. Soon you'll be making your own fluorescent-conjugated antibodies and doing IFA.
Great shots.

David

Jacek · Post by **Jacek** » Mon Dec 12, 2016 2:25 pm

Bravo. SUPER !

zzffnn · Post by **zzffnn** » Mon Dec 12, 2016 3:46 pm

Very beautiful! Thankbyou for sharing!

carlos.uruguay · Post by **carlos.uruguay** » Mon Dec 12, 2016 7:07 pm

Very nice!!!

Sumguy01 · Post by **Sumguy01** » Mon Dec 12, 2016 9:03 pm

Nice photos.
Very interesting also.
Thanks for sharing.

harisA · Post by **harisA** » Tue Dec 13, 2016 4:59 am

very nice images.Is this with epi fluorescence illumination?

Pau · Post by **Pau** » Tue Dec 13, 2016 7:20 am

Thanks all for your kind comments!

discomorphella wrote:...Soon you'll be making your own fluorescent-conjugated antibodies and doing IFA.

Not likely to happen!, I reserve those techniques for researchers at well founded labs. Maybe I could borrow some Arabidopsis samples expressing GFP, not produced at home of course

Haris, yes, it's standard epifluorescence.

discomorphella · Post by **discomorphella** » Tue Dec 13, 2016 10:09 pm

I don't know Pau, you're a fearless optical experimenter, I have no doubt you could raise some immunoglobulins in rabbits for example, and FITC is pretty easy to get...I am sure you could do it. You are correct that getting a lot of different colors for IFA will get expensive, since all the nicer conjugating dyes are not cheap, and one gets tired of seeing only FITC...but I have no doubt you could do it. On the other hand, as Igor has shown, Congo Red, Calcofluor White and Acridine Orange can make some spectacularly nice photomicrographs...

Best,

David

hkv · Post by **hkv** » Thu Dec 15, 2016 2:22 pm

Very nice! Thank for sharing. Seems like your new rig is delivering!

RogelioMoreno · Post by **RogelioMoreno** » Thu Dec 15, 2016 2:43 pm

Very nice set!

Rogelio

WalterD · Post by **WalterD** » Tue Dec 20, 2016 11:41 am

Very nice set of photographs Pau.(':smt041')
The 2nd one has got that "blacklight" effect. Tthanks for the detailed description of your new fluorescent illumination setup, impressive indeed. Would be interesting to have a shortlist of objects that are autofluorescent by nature, as far as I understood it's a bit trial and error to find out which are.

fpelectronica · Post by **fpelectronica** » Thu Dec 22, 2016 11:53 am

Very nice!