Auto Fluorescence + 3D Deconvolution - The Beauties (part 1)

RogelioMoreno · Post by **RogelioMoreno** » Tue Sep 24, 2013 4:07 am

Here are the first group of the beauties.

All images were done with the Plan Apo 20x/0.75 and the UV-2B cube with auto fluorescence. You have to do the stack quicly because the specimen auto-fluorescence get dim with the pass of the time.

Closterium by Rogelio Moreno G., on Flickr

Micrasterias laticeps by Rogelio Moreno G., on Flickr

Micrasterias furcata by Rogelio Moreno G., on Flickr

Micrasterias radiosa by Rogelio Moreno G., on Flickr

Rogelio

Fredlab · Post by **Fredlab** » Tue Sep 24, 2013 4:13 am

arf...

speechless

Cactusdave · Post by **Cactusdave** » Tue Sep 24, 2013 4:41 am

Fascinating and beautiful results. Alas this could be yet another new way to drain the wallets of long suffering photomicrographers!

Marek Mis · Post by **Marek Mis** » Tue Sep 24, 2013 5:28 am

Rogelio,

Really spectacular images ! All of them are beautiful but my favourite is the 4 th one.
Thanks for sharing.

Marek

Jacek · Post by **Jacek** » Tue Sep 24, 2013 5:30 am

beautiful

Ernst Hippe · Post by **Ernst Hippe** » Tue Sep 24, 2013 6:10 am

Rogelio,
phantastic pictures of my beloved desmids!
The M.laticeps is not common over here - I got it only once from a supplier of tropical water plants, and only the variation with single tips at the side lobes. Where from did you get it and which variation(s)?
Regards Ernst

Ecki · Post by **Ecki** » Tue Sep 24, 2013 6:58 am

Rogelio,

this is amazing. I think I need deconvolution software as well

What package did you use?

Best regards
Ecki

discomorphella · Post by **discomorphella** » Tue Sep 24, 2013 8:10 am

Beautiful results. Now you need to figure out which antifade reagents are the least damaging to algae. You might try some dilute p-Phenylenediamine or DABCO (start with perhaps <= 20 micromolar in the water you use to mount the specimens and work up to larger concentrations if required) to try and reduce the photobleaching. Deconvolution results look great, can you please include some details as to which algorithm, how many slices, which model parameters etc that you are using?

David

Pau · Post by **Pau** » Tue Sep 24, 2013 8:26 am

Extraordinary quality, both this ones and your Spirogyra.

Litonotus · Post by **Litonotus** » Tue Sep 24, 2013 8:29 am

I'm sure we will see one of them in top 10 of small world or bioscapes (: great!

pwnell · Post by **pwnell** » Tue Sep 24, 2013 11:57 am

4th one is in a class of its own.

arturoag75 · Post by **arturoag75** » Tue Sep 24, 2013 12:55 pm

Really stunning and fascinating imsges.........
I' d like to know more on this tecnic

sebba28 · Post by **sebba28** » Tue Sep 24, 2013 2:54 pm

Stunning pictures,

RogelioMoreno · Post by **RogelioMoreno** » Wed Sep 25, 2013 3:48 am

Thank you for your comments.

Ernst, I am getting my samples from lake Miraflores (this lake has some degree of salt because it is a reservoir for the Panama canal). If the specimen on picture #4 has double tips on the side lobes then I am finding single and double.

Ecki, I am using the 3o days trial of cellSens and AutoQuant X3.

David thank you for the tips about the p-Phenylenediamine, do you know where I can get it?

I am using the default settings on both cellSens and AutoQuant 3D deconvolution, no SA detection.

#1: 2322 x 1179 x 24 slides
#2: 2498 x 1339 x 14 slides
#3: 1851 x 1750 x 20 slides
#4: 1684 x 1688 x 23 slides
#5: 2061 x 2010 x 21 slides
#6: 2161 x 2073 x 16 slides

All with the Plan Apo 20x/0.75 and 1 um step.

Rogelio

Ecki · Post by **Ecki** » Wed Sep 25, 2013 4:11 am

Hi Rogelio,

I looked at AutoQuant X3 and was very impressed. Do you know how much it costs?

Regards
Ecki