Jump to content
ReefCleaners.org

Mr. M's Microscopy of the Reef, FlowerMama's Dinos! #303


Mr. Microscope

Recommended Posts

  • 3 weeks later...
  • Replies 337
  • Created
  • Last Reply
Mr. Microscope

Hello All,

As some of you know, I lost my sea urchin during a transition back at the end of August. I was finally able to put forth a fitting memorial to it in the form of electron microscopy. Here it was in all it's reefy glory.

Echinometra Viridis
Urchin0120130225_zps07ff1938.jpg

In the above pic, you can see little lines in the spines. Here they are up close.
Spine02_zpsacbb5837.jpg

Spine01_zps40feb370.jpg

I also looked at the spine in cross section.
Spine04_zpsbefe55fc.jpg

Check out how the bone is built up with a face-centered hexagonal pattern.
spinefchex_zps1d82d417.jpg

Additionally, I took a look at the body shell. It is composed of many nodules where the spines connected to the body and could rotate freely like a knee or elbow joint.
Shell01_zpse19a3085.jpg

Holes lined the shell where podia would protrude.
Shell02a_zps821097af.jpg
The resolution here really doesn't do the pic justice. Here is a link to download an ultra-high resolution .tif of this image if you're interested. It should look pretty awesome on one of those new screens. Resolution is 5120 x 3560. The download is about 20MB.
https://dl.dropboxusercontent.com/u/6760404/Shell%2002%20UHD.tif

Finally, here is a video I made a while back featuring this urchin.

Link to comment
  • 1 month later...
Mr. Microscope

Dude, this thread is so freaking cool.

Thank you Dryland! I don't get to update it nearly as frequently as I'd like.

 

 

That is pretty sweet!!!!!

Thank you Chris!
Link to comment
  • 1 month later...
AllOutAfrican

wow this thread is so interesting its amazing to see a coral so close up thanks for taking the time to do this.. i havent had time to read through it all but will defiantly read through..

Link to comment
Mr. Microscope

wow this thread is so interesting its amazing to see a coral so close up thanks for taking the time to do this.. i havent had time to read through it all but will defiantly read through..

Thank you AllOut! I'm actually about to update it again. Stay tuned. :)

Link to comment
Mr. Microscope

Hello All!

Many of you may have seen Charlie Mazel's MACNA speech. If not, here it is:

https://youtu.be/3rC4G84OWIg

Charlie is the guy behind the NightSea fluorescence viewing system. He's pretty much responsible for the new passion for so-called, fluorescence night diving, and also markets systems to adapt pretty much any stereo microscope for fluorescence. Yesterday, I had the good fortune to have Charlie come to my lab and do a demo of his fluorescence systems on one of my microscopes.

Charlie%20Mazel_zpscxzsuheo.jpg
(No, we did not plan to coordinate outfits. lol :P )

Here's our microscope retrofitted with the NightSea system.
Leica%20M80%20with%20NightSea_zpscx1wyrc

 

Here's Dr. Mazel explaining the system:

https://youtu.be/ucPX4qsQQww


One of my bubble tip anemones recently split, but it did so somewhat unevenly. I was left behind with one of the smallest clones I've ever seen. It's no larger than a quarter. So, I figured it would be a great candidate to image with this system. I carefully took it out of the basket in my tank and brought it to my facility for the day.

 

Here's a picture of the mother before it split. This was taken under an estimated 12-14K.
nem07_zpssce9g62o.jpg

 

There are several lampsets/filters available for the system and Charlie brought the royal blue lamp to stimulate green fluorescence, a cyan lamp for yellow-orange fluorescence, and a green lamp for red fluorescence. I managed to image the BTA under all three conditions. Please forgive any blurriness as I was only able to get pictures by holding my Canon point-and-shoot camera up to one of the oculars of the microscope.

From left to right: RB, Cyan, and Green source.
Three%20Emissions%2002_zpswifltxo9.jpg

 

Here's the lamp heads and corresponding filters starting with UV, RB, Cyan, and Green.
sfa10.jpg
The filters (Yellow, orange, and red) cut out signal from the LED that would obscure the image. Unlike taking pictures of our corals under blue LED's and having the camera blown out by blue, the filter removes the blue so that we are only left with the true fluorescent signal.

The figure below shows the effect of filtering with the cyan source. The image on the left is unfiltered. You can clearly see the the effects of fluorescence as the cyan source excites the proteins of the anemone, but the picture is obscured by the cyan background and effects of cyan light on the fluorescent signal. The image on the left has been filtered. Notice, we still see the fluorescent signal from the BTA, but the background is completely devoid of cyan.

unfiltered%20and%20filtered%20YFP_zpsph1

 

It was amazing how the green fluorescent proteins (GFP) lit up. This BTA was of the speckled variety. Here, we get to see the intense emissions from these speckles made up of cells saturated with GFP.

GFP%20Speckle_zpshkcbldef.jpg

 

This image was taken utilizing the green LED source to excite red fluorescent proteins (RFP).

RFP%2002_zps7pg1nwjr.jpg

 

Looking closer with the royal blue emission, we were able to see how the orange and yellow appearance to the tentacles is actually composed not of orange or yellow pigments, but of a coalescing of GFP and RFP in a graded series of concentrations. Using Photoshop, I isolated the green and red signals so you can observe their overlap to create the rainbow effect we see in our "Rainbow Bubble Tips Anemones."

GFP%20and%20RFP_zpsqsd8srit.jpg

 

Here, closer to the base of the tentacle, you see an individual cell lit up with GFP.

Individual%20GFP_zpsh3vhpkoi.jpg

 

To exit, just one more shot of the BTA's under all three conditions. These pics were not taken under the microscope, but with viewing shield filter I placed in front of my camera.

Three%20Emissions%2001_zpsg0iaa7sh.jpg

Click to view larger.

 

I had a great time with Charlie and I hope that our facility will be purchasing one or two of these systems in the near future. This tool will definitely be useful for projects from biology and cancer research, to hard and soft materials science and engineering. I was truly inspired by this experience. I'm ready to go find a huge yellow screen to put in front of my tank for night viewing. lol

Link to comment
Mr. Microscope

Very cool write up. Thanks for sharing!

Thank you for checking it out Slow!

 

Love this thread and all the work you have done. Awesome

Thank you Maniu! It's a lot of fun!

 

Yep. This made my day. Awesome blend of photography and science!

Awesome! Thank you neuwave!
Link to comment
Mr. Microscope

I'm going to try some micro CT X-ray, I think I can get down to .35 microns. It's good to be a geek.

Cool cool! Pretty stellar resolution on that. What are you imaging?
Link to comment
Mr. Microscope

Last week it was mice femur's for bone density. Not my main gig but entertaining.

Neat!

What is your main project?

Link to comment

I'm an engineer mostly installing Nuclear Magnetic Resonance Spectrometers. Occasionally I get to play with something interesting at the universities I visit.

And lest I forget I'd like to compliment you on your imagery. I believe I caught some of Charlie's speech on youtube and plan on trying some filters to get the blue out of my pictures.

Link to comment
Mr. Microscope

I'm an engineer mostly installing Nuclear Magnetic Resonance Spectrometers. Occasionally I get to play with something interesting at the universities I visit.

And lest I forget I'd like to compliment you on your imagery. I believe I caught some of Charlie's speech on youtube and plan on trying some filters to get the blue out of my pictures.

Cool! We have a couple of those here at Northwestern. I can actually see them from my office.

Yeah, I usually just use a white balance, but this has me rethinking things.

 

:eek: :eek: :eek: :eek:

 

:bowdown: :bowdown: :bowdown: :bowdown:

 

That is all

:blush: Thank you PC! :D

 

Haha! I was just trying to reply here and look what popped up! I guess 10 is the max.

Screen%20Shot%202015-03-12%20at%209.47.3

 

 

 

Google confirmed, these are real words

LOL!
Link to comment
Polarcollision

Cool! We have a couple of those here at Northwestern. I can actually see them from my office.

Yeah, I usually just use a white balance, but this has me rethinking things.

 

:blush: Thank you PC! :D

 

Haha! I was just trying to reply here and look what popped up! I guess 10 is the max.

Screen%20Shot%202015-03-12%20at%209.47.3

 

 

 

LOL!

 

Yes it it, I tried to put a whole row of each! Phtttbt. limiting our praise, ;-)

Link to comment

Archived

This topic is now archived and is closed to further replies.

  • Recommended Discussions


×
×
  • Create New...