Polypodium sporangia fluorescence

Images made through a microscope. All subject types.

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micro_pix
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Polypodium sporangia fluorescence

Post by micro_pix »

These are the sporangia of the fern Polypodium vulgare auto-fluorescing under UV. I immersed the leaf in water and used an Olympus 10x water dipping objective. I liked the way that the spores are visible inside the sporangia.

Dave
Polypodium vulgare sporangia
Polypodium vulgare sporangia
Polypodium vulgare sporangia
Polypodium vulgare sporangia
Last edited by micro_pix on Thu Nov 12, 2020 1:55 pm, edited 2 times in total.

Pau
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Re: Polypoium sporangia fluorescence.

Post by Pau »

Excellent pictures of one of my favorite subjects :D
What UV source and filters did you used?
Pau

micro_pix
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Re: Polypoium sporangia fluorescence.

Post by micro_pix »

Pau wrote:
Thu Nov 12, 2020 12:41 pm
Excellent pictures of one of my favorite subjects :D
What UV source and filters did you used?
Thanks Pau. I used my infinite Olympus BH2 with the standard mercury lamp and UV dichroic mirror - no extra barrier filter. The objective was an Olympus UMPlanFL 10x W

Dave

Lou Jost
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Re: Polypodium sporangia fluorescence

Post by Lou Jost »

Great use of the water objective. How did you avoid air bubbles? I tried this once but the bubbles ruined it.

micro_pix
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Re: Polypodium sporangia fluorescence

Post by micro_pix »

Lou Jost wrote:
Thu Nov 12, 2020 2:10 pm
Great use of the water objective. How did you avoid air bubbles? I tried this once but the bubbles ruined it.
It’s the first time I’ve used a water objective so I didn’t have any experience. I must have been lucky; I just put it in a petri dish, weighted the ends of the leaf with a couple of steel washers and filled the dish with de-ionised water. I’m not sure if the de-ionised water makes a difference with bubbles but I always use it for microscopy.

Dave

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Re: Polypodium sporangia fluorescence

Post by Lou Jost »

Thanks, maybe that's the key.

Sym P. le
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Re: Polypodium sporangia fluorescence

Post by Sym P. le »

Thanks for posting. It adds a whole new level of understanding for these sporangia I keep finding laying around. :wink:
Attachments
Mite with Sporangia.jpg

Sumguy01
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Re: Polypodium sporangia fluorescence

Post by Sumguy01 »

=D> Very nice.
Thanks for sharing.

micro_pix
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Re: Polypodium sporangia fluorescence

Post by micro_pix »

Lou Jost wrote:
Thu Nov 12, 2020 4:37 pm
Thanks, maybe that's the key.
Possibly the de-ionised water has a lower surface tension than tap water because of a lack of impurities, it certainly seems to flow more freely on a surface, which would help reduce bubbles.

Dave

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Re: Polypodium sporangia fluorescence

Post by Pau »

micro_pix wrote:
Fri Nov 13, 2020 2:12 am
Lou Jost wrote:
Thu Nov 12, 2020 4:37 pm
Thanks, maybe that's the key.
Possibly the de-ionised water has a lower surface tension than tap water because of a lack of impurities, it certainly seems to flow more freely on a surface, which would help reduce bubbles.

Dave
It could be, although I lean to think more about the composition and structure of the plant surface.
Most plants have hydrophobic substances like wax in their surfaces to impermeabilize them in order to avoid excessive water evaporation, this effect can be enhanced with small hydrophobic hairs.
Plants living in humid environments like ferns often have less hydrophobic surfaces.

Maybe adding an humectant substance to the water could aid to avoid bubbles, I'm thinking in a product used to improve uniform drying of photographic paper (old times dark room memories :) ) or dish washer brightener. Liquid soap also would do but likely it will be more aggressive
Pau

Lou Jost
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Re: Polypodium sporangia fluorescence

Post by Lou Jost »

Maybe adding an humectant substance to the water could aid to avoid bubbles
This is definitely true, and when I am shooting through water with a normal objective, I always use it. This also reduces meniscus effects that can distort the image, and I suspect it also reduces wave activity due to vibrations. But I am afraid to use a surfactant with a water-dipping objective, because it might affect the seals or leave a residue.

micro_pix
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Re: Polypodium sporangia fluorescence

Post by micro_pix »

I have occasionally used a tiny amount Decon90 (a strong surfactant) when photographing fungal structures in water under a coverslip and that helped avoid bubbles but I haven’t used it lately. A very unscientific observation I know but a drop of de-ionised water does seem to spread out and wet the surface of a fern leaf much better than a drop of tap water and I was surprised at how well it displaced the air around the nooks and crannies of the sporangia.

Edit: I have sought out some reliable sources and it appears that de-ionised water has a higher surface tension than tap water! Well I did say my observation wasn’t scientific.

Dave

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Re: Polypodium sporangia fluorescence

Post by Macro_Cosmos »

These certainly look alien and spooky to me, I love it! One day, I'll get that expensive bx3-ura or DIY a cage system to do fluorescence.

Lou Jost
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Re: Polypodium sporangia fluorescence

Post by Lou Jost »

I do fluorescence with no filter cube, very cheap.

micro_pix
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Re: Polypodium sporangia fluorescence

Post by micro_pix »

Did you show this in a previous post Lou? If you did could you post a link, I’d like to see how you have set it up.

Dave

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