Some autofluorescence

Images made through a microscope. All subject types.

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sushidelic
Posts: 233
Joined: Fri Jun 10, 2016 5:46 am

Some autofluorescence

Post by sushidelic »

Hello!

Optolong was so kind to send me a sample of their Longpass DAPI Single Band Filter Set for testing purposes, and I am really happy with the results. It yields brighter fluorescence than my U-MWU2 from Olympus, which is similar in specs, but might be a bit aged already as excuse.
I went through my garden to try it on some autofluorescence targets and did some stacks of the stellate leaf hairs on the epidermis of a Deutzia scabra, farn sporangia and a moss sporangium.
Done on a Olympus BX52 with UMPlanFL objectives, Camera was a Vieworks VA47MC, custom tube lens.

Image

Image

Image

Hope you like it!
Best regards,
Michael

iconoclastica
Posts: 393
Joined: Sat Jun 25, 2016 12:34 pm
Location: Wageningen, Gelderland

Re: Some autofluorescence

Post by iconoclastica »

I have been looking at fern indusia for the better part of the week, but these I didn't even recognize as such - which is great!
=D>
--- felix filicis ---

sushidelic
Posts: 233
Joined: Fri Jun 10, 2016 5:46 am

Re: Some autofluorescence

Post by sushidelic »

Thank you!

The indusiae of the farn were still completely closed. It was surprising, that the fluorescence of the interior was so clearly visible, refracted by the transparent hull.

Edit - Sorry, confused it with the moss!
The Moss was very old and dry when I took the pictures, so maybe quite some decay involved ;)

micro_pix
Posts: 469
Joined: Fri May 11, 2012 12:05 pm
Location: Southampton, Hampshire, UK

Re: Some autofluorescence

Post by micro_pix »

Very nice!

Dave

Adalbert
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Joined: Mon Nov 30, 2015 1:09 pm

Re: Some autofluorescence

Post by Adalbert »

very nice :smt041

iconoclastica
Posts: 393
Joined: Sat Jun 25, 2016 12:34 pm
Location: Wageningen, Gelderland

Re: Some autofluorescence

Post by iconoclastica »

With this photo I am sure youwould win next years photo contest of the British Pteridological Society :P
Is it the DAPI filter set specifically that makes it work so well?
--- felix filicis ---

Luis Carlos
Posts: 75
Joined: Wed Oct 30, 2019 3:42 am

Re: Some autofluorescence

Post by Luis Carlos »

Beautiful pictures!

Lou Jost
Posts: 5091
Joined: Fri Sep 04, 2015 7:03 am
Location: Ecuador
Contact:

Re: Some autofluorescence

Post by Lou Jost »

Really nice work, especially the moss image. Like Iconoclastica, I would love to learn more technical details. I am curious about the "custom tube lens". WHat was its focal length? Also, what an amazing camera. I imagine it is very expensive.

sushidelic
Posts: 233
Joined: Fri Jun 10, 2016 5:46 am

Re: Some autofluorescence

Post by sushidelic »

Thank you!

Now I learned that there is a British Pteridological Society - at first glimpse I thought this might be something Monty Python related ;)!

The Filter set itself is not really "special", it is basically a UV exitation / long pass filter. Here are the specs: https://www.optolongfilter.com/en/produ ... ilter-sets
But I was really happy with the transmission, the fluorescence was very bright and vivid.

Due to the laaaarge sensor of the camera, I removed the trinocular head of the BX52 and placed a 300mm Apo Gerogon as tube lens instead. This nearly covers the full sensor now, but Image quality towards the edges decreases and certainly vignetting is visible. I mainly used the cam, because I can let it run continuously and record while turning the fine focus, no shutter shock or anything. Quality wise probably not the best solution due to the continuous motion, but the results look ok, so I'm good.

Those industrial Camera Link CCDs used to be madly expensive, but with some patience and luck they show up for reasonable prices on the second hand market. Getting them to run is quirky, as Camera Link is not comparable wit USB in terms of ease of use.

Best regards,
Michael

Sumguy01
Posts: 1647
Joined: Mon Jan 28, 2013 11:05 pm
Location: Ketchikan Alaska USA

Re: Some autofluorescence

Post by Sumguy01 »

=D> Very nice.
Thanks for sharing.

Pau
Site Admin
Posts: 5470
Joined: Wed Jan 20, 2010 8:57 am
Location: Valencia, Spain

Re: Some autofluorescence

Post by Pau »

Very good images!

What's the sensor size? (googling Vieworks VA47MC only shows your images at facebook)
Is the vignette due to the fluorescence illuminator or to the microscope image path?
Pau

sushidelic
Posts: 233
Joined: Fri Jun 10, 2016 5:46 am

Re: Some autofluorescence

Post by sushidelic »

Hi Pau,

thank you, here are the specs of the chip, an On Semi KAI-47051

Number of Active Pixels: 8856 (H) ×5280 (V)
Pixel Size: 5.5µm (H) ×5.5µm (V)
Active Image Size: 48.7 mm (H) × 29.0 mm (V), 56.7 mm (diagonal)

Tube lens of a common Oly Infinity system is 180mm, official image circle 26.5mm.
So with a 300mm tube lens, my supported field should be 44mm. Vignetting in the corners is mostly due to Epi Fluorescence, fiddling with the lamp position and collimator settings helps a bit. With transmitted illumination, vignetting is far less.

Best regards,
Michael

EDIT: here are some links regarding the camera
https://www.stemmer-imaging.com/media/u ... -M-C-7.pdf
https://www.onsemi.com/pdf/datasheet/kai-47051-d.pdf

ModelZ
Posts: 49
Joined: Sun Nov 01, 2020 4:34 am
Location: Northern Europe

Re: Some autofluorescence

Post by ModelZ »

Very nice work, beautiful palette & detail.
Vigneting is easy to fix in the post. You have just a problem of too good a camera ;-)

seta666
Posts: 1062
Joined: Fri Mar 19, 2010 8:50 am
Location: Castellon, Spain

Re: Some autofluorescence

Post by seta666 »

This is nice!!!, I would be interested in the set up needed to get this auto fluorescence and add it to my BHT

OptolongSherry
Posts: 2
Joined: Mon Jul 05, 2021 8:10 pm
Location: China

Re: Some autofluorescence

Post by OptolongSherry »

Dear Mr.Michael

Thank you so much for post tested images here,really apprecaited for your help to shinning our brand here!
We are still looking for someone who could be testing our filter set and sharing testing images on social media or make videos:
We can currently provide:
1) Single band pass: DAPI, FITC, TRITC, TexasRed, Cy5
2) Double Band Pass: FITC /TRITC,Green/Orange,Aqua/Green/Orange
3) FISH app: FISHAqua, FISHGreen, FISHOrange, FISHRed.
4) Long Wave pass series: UV-DAPI, B,G,V
5) And customized fluorescent filters

If there are someone who intetrested to testing,email to lens@optolong.com
Sherry Yan | Optical Coating Filter (Manufacturer)
Optolong Optics Co.,ltd | Room 1523-1526 Kepu Road | Kunming City | Yunnan Province | China 650102
WhatsApp:18388158376 | Skype:Sherry Yanfilter
| lens@optolong.com | www.optolongfilter.com | www.optolong.com

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