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Lighting for macro photography of fern gametophytes
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jsp



Joined: 28 Mar 2015
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PostPosted: Fri Mar 16, 2018 11:53 pm    Post subject: Reply with quote

That's lovely :-) Thanks for showing me. :-) My setup only has three parameters that can be changed though, so it's probably safer just to stick with the arduino program and avoid introducing any bugs.

This is my setup:

https://github.com/BioMakers/23_Focus-stacking-system-for-gametophyte-ferns/blob/master/README.md
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jsp



Joined: 28 Mar 2015
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PostPosted: Fri Jun 08, 2018 3:46 am    Post subject: Reply with quote

Hello :-)

I have been to visit a collaborator at the Sainsbury Laboratory here in Cambridge and I photographed some of his Arabidopsis leaf hairs (trichomes). The trichomes are very glass-like and the compilation in Helicon focus hasn't worked very well, I think because of diffraction. I wondered if anyone might have an idea of where I am going wrong? I also did not get my black background right, so I think I need to work on that.

Thanks!



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ChrisR
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PostPosted: Fri Jun 08, 2018 4:06 am    Post subject: Reply with quote

Try the other Helicon modes. Can' t remember which - in Zerene it would be Pmax. You would expect to have to retouch between two stacks.
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PostPosted: Fri Jun 08, 2018 4:31 am    Post subject: Reply with quote

Thanks!
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jsp



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PostPosted: Fri Jun 08, 2018 4:39 am    Post subject: Reply with quote

This is it redone with weighted average. The other one was depth map. I have one more to go but have to dash out. Thanks! This looks better already. Phew!
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PostPosted: Fri Jun 08, 2018 6:27 am    Post subject: Reply with quote

This is the pyramid method.

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PostPosted: Fri Jun 08, 2018 6:29 am    Post subject: Reply with quote

If I am just trying to capture the shape of the trichome, do you think it would be better to go to higher magnification and just take the one trichome? That was 10x, but I now have the option of 20x and 50x.
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PostPosted: Fri Jun 08, 2018 6:34 am    Post subject: Reply with quote

This is better if I just take enough slices to get the one trichome in focus. I think if I put a black background on that it might be nice.

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PostPosted: Fri Jun 08, 2018 8:34 am    Post subject: Reply with quote

It looks a lot better cropped. Maybe this is the composition I should be aiming for and maybe I should use a higher magnification lens. I will ask the plant's owner if that would work for him.
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PostPosted: Tue Jun 12, 2018 6:42 am    Post subject: Reply with quote

I went back today for another go and got these better images. It turns out that my collaborator didn't want close-ups of the trichomes, but rather wanted the whole leaf, so these worked out nicely with the 5x objective. That was the first time I'd used it. I used a 120 micron step size.









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PostPosted: Tue Jun 12, 2018 7:10 am    Post subject: Reply with quote

Nice, but some of those single leaf stacks should have been deeper. The trichome spikes closest to the camera are not in focus, but could have been if you had started the stack higher up. When you are determining the limits of the stack, it is very easy to miss those kinds of details. It helps to look the output on a larger monitor.
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jsp



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PostPosted: Tue Jun 12, 2018 9:44 am    Post subject: Reply with quote

Hi Lou,

Thanks yes we noticed that. My collaborator was happy to get the work done fast and not worry about losing the tips, so I went with his requirements rather than take up more of his time.

I was pleased with the focused bits though. :-)
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PostPosted: Tue Jul 31, 2018 2:59 pm    Post subject: Reply with quote

Hi,

I just got an email from someone who wants to make a proper film about fern gametophytes. I'm interested to help and have written back, but I just wondered if I could mention here in case anyone else would like to contribute?

This is the email:

(Please PM if you would like the email address to reply to. )

I have been recommended by Andrew Leonard to get in contact with you regarding a short documentary I am currently producing. I am a student at the National Film and Television School and am currently working on a film about the alternation of generations in ferns, including its evolutionary implications and placement in the timeline of land plants. I am looking for some help with fact consolidation but also for access to archive footage or a microscope and a fern gametophyte to demonstrate the complexity and scale of the different life stages. I understand these may be unrealistic or difficult demands to meet, and that you may not have the time for such a project, but the films from my school often get a high degree of attention and I am hoping the same may go for my film – if not just to educate people in an entertaining way.
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