Some autofluorescence
Moderators: rjlittlefield, ChrisR, Chris S., Pau
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- Joined: Fri Jun 10, 2016 5:46 am
Some autofluorescence
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.
Hope you like it!
Best regards,
Michael
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.
Hope you like it!
Best regards,
Michael
- iconoclastica
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- Location: Wageningen, Gelderland
Re: Some autofluorescence
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!
--- felix filicis ---
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- Joined: Fri Jun 10, 2016 5:46 am
Re: Some autofluorescence
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
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
Re: Some autofluorescence
Very nice!
Dave
Dave
Re: Some autofluorescence
very nice
- iconoclastica
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- Location: Wageningen, Gelderland
Re: Some autofluorescence
With this photo I am sure youwould win next years photo contest of the British Pteridological Society
Is it the DAPI filter set specifically that makes it work so well?
Is it the DAPI filter set specifically that makes it work so well?
--- felix filicis ---
- Luis Carlos
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- Joined: Wed Oct 30, 2019 3:42 am
Re: Some autofluorescence
Beautiful pictures!
Re: Some autofluorescence
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.
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Re: Some autofluorescence
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
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
Re: Some autofluorescence
Very nice.
Thanks for sharing.
Thanks for sharing.
Re: Some autofluorescence
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?
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
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- Joined: Fri Jun 10, 2016 5:46 am
Re: Some autofluorescence
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
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
Re: Some autofluorescence
Very nice work, beautiful palette & detail.
Vigneting is easy to fix in the post. You have just a problem of too good a camera ;-)
Vigneting is easy to fix in the post. You have just a problem of too good a camera ;-)
Re: Some autofluorescence
This is nice!!!, I would be interested in the set up needed to get this auto fluorescence and add it to my BHT
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- Location: China
Re: Some autofluorescence
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
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