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


Mr. Microscope

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Don't mean to sidetrack the thread but I have another Micro CT to install next month I wonder what a Zooanthid polyp looks like in 3D. If I can can work the logistics and figure out how to keep it alive and possibly open that would be great. Never would have thought about adding more science to reefkeeping without this thread. As I said, it good to be the Geek.

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Mr. Microscope

Don't mean to sidetrack the thread but I have another Micro CT to install next month I wonder what a Zooanthid polyp looks like in 3D. If I can can work the logistics and figure out how to keep it alive and possibly open that would be great. Never would have thought about adding more science to reefkeeping without this thread. As I said, it good to be the Geek.

I don't know about alive, but some glutaraldehyde may fix it in an open state (might even open it up further). I'd try processing it with a graded series of saltwater to fixative, eventually ending up with 4% Paraformaldehyde and 2.5% Glutaraldehyde. Or you could just try adding the fixative straight to the saltwater and make your end concentration equal to the aforementioned formula. Would you be able to image it while still submerged?
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No problem I could image it in a small vial of salt water centered in the scanner. I'm sure I can acquire the sample prep materials at the university if the customer agrees it would be a good test of the system.

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Mr. Microscope

No problem I could image it in a small vial of salt water centered in the scanner. I'm sure I can acquire the sample prep materials at the university if the customer agrees it would be a good test of the system.

Cool! I'd mount it in the vial and let it heal open for a while until it opens up regularly in your tank. Then, take it to the lab and empty out half of the vial. Expose it to intense enough lighting for it to open up again. Once it's fully open, slowly add a 2x concentration of fixative to the vial so that you end up with the correct concentration in the end. You probably need to make the fixative with salt water so you don't suffer any osmotic effects.
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Coral Scaper

Awesome thread!

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

image_12994.jpg

;)

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Mr. Microscope

Keyence brought in one of their microscopes to demo in my lab today. This is probably one of the fanciest stereo microscopes I've ever seen. Everything about it was motorized and digitized. It can do some really amazing things including building up 3D renderings of your sample. The microscope scans thought the entire focal length of the sample and generates an image where everything is in focus. This is impossible to do with a normal microscope where maybe 5-10% of your image would be in focus.

optical-microscope-3d-4980-5799473.jpg

 

I looked at my brittle starfish sample, my sea urchin sample, and my old montipora digitata skeleton. I'll start with a little follow-up on my microscopy of montipora digitata from earlier in the thread. Here's a link in case you haven't seen it yet:

http://www.nano-reef.com/topic/280413-mr-microscopes-microscopy-of-the-reef-nightsea-262/?p=3584246

 

Check it out, these images are completely in focus.

VHX_000050_zps4qie05tc.jpg

 

VHX_000052_zpslgxxms9y.jpg

 

photobucket is being finicky right now. I'll upload more later when I get it working. This is just a tease. I have some really cool 3D stuff coming up..

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Mr. Microscope

Okay. So, here's the Urchin stuff.

again, here's a link to the previous microscopy:

http://www.nano-reef.com/topic/280413-mr-ms-microscopy-of-the-reef-keyence-284/?p=4905161

 

Here's what the piece of skeleton looked like that I put in the electron microscope. Did I mention, this Keyence can tilt in all directions?

VHX_000059_zpskyrbwrll.jpg

 

The microscope also has a feature to automatically gather high dynamic range information (HDR) which is especially useful for very white or reflective surfaces like this. Here's the sample with normal light Vs. HDR.

Normal

VHX_000057_zpsfczpfnys.jpg

 

HDR

VHX_000058_zpsywaiqtpk.jpg

 

We zoomed in on one of the needle joints:

VHX_000056_zpshuydhfba.jpg

 

Using the focus gradient, we were able to capture a profile and render some 3D data:

VHX_000053_zpsmcisqetb.jpg

 

You can see that joint is about 1mm tall.

Rotated:

VHX_000054_zpsn0sabrut.jpg

 

The AOI was cropped to show just the joint:

VHX_000055_zpsgyhipyoc.jpg

 

Finally, we looked at one of the needles. Here it is in cross section. Again, this is not a flat surface. Without the focus blending we wouldn't be able to see 10% of this detail.

VHX_000069_zps9hocjhdp.jpg

 

You can see the dips and pits in the needle on its side here in 3D. We measured the depth at about 35um.

VHX_000070_zpsdfudo8ql.jpg

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Mr. Microscope

That's a pretty awesome stereomicroscope! Love this!

Thank you Felicia! One day, I'll have to compile all of this into a book or website.

 

Amazing photos. Well done, Sir! Nature at its most beautiful B)

Thank you Slow! Nature is my super model. lol

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Mr. Microscope

This thread is unbelievable. The pictures are amazing, thank you Mr. Scope.

 

Thank you Kat! I have to live up to my name, right? lol

I don't get to update it nearly as often as I'd like. Slow and steady wins the race I guess. ;)

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FlowerMama

Impressive! I've never seen images like that. Beyond words amazing. Keep them coming!!!! Now I have to back up and remind myself of other pics I saw before but forgot about, like the amphipod head-on shot.

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Mr. Microscope

sooo interesting!

Thank you M!

 

Impressive! I've never seen images like that. Beyond words amazing. Keep them coming!!!! Now I have to back up and remind myself of other pics I saw before but forgot about, like the amphipod head-on shot.

Thank you Annette! Yes, they range from beauty to nightmare-inducing. lol

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Scanning Electron Microscopy of ZooplanktonHello All!It's taken me a while to gather all of this data; a minute here and there after sessions of looking at actual research samples, but I've finally managed to gather a decent collection of images. This entry's samples consist of some pods that I managed to get out of my fuge. I found an infant amphipod, a copepod, something that looks vaguely like a caterpillar, and something that may be a microscopic bivalve of sorts. Enough chatter, here's the good stuff!First off, here is the amphipod I found. I used some static to get him to stand on his hind legs so that I could look him straight in the eye.

File02b.jpg

ROOOAR!!!

File05b.jpg

Actually, I say he looks a little like Tony the Tiger. What do you think? :lol: :lol:

FU8184lg.jpg

Let's zoom in on that mouth a little.

File41b.jpg

"The better to EAT you with!!!"File42b.jpg

Arms:

File45b.jpgThe CLAW!File47b.jpg

File13b.jpg[/center]Next, the bivalve thing. My other guesses are that it is a protozoan, daphnia, or maybe cladoceran Evadne sp. Again, please contact me on this one if anyone knows for sure what this is.

File15b.jpg

Zooming in on the edge.

File19b.jpgFile30b.jpg

pretty old thing for me to be quoting but that is pretty cool, you should get one on a fully developed adult amphipod eventually. also that bivalve of yours looks a bit like an ostracod, some species are hairy on the shells, I could be wrong though.
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Mr. Microscope

pretty old thing for me to be quoting but that is pretty cool, you should get one on a fully developed adult amphipod eventually. also that bivalve of yours looks a bit like an ostracod, some species are hairy on the shells, I could be wrong though.

Thank you amphipod! Believe it or not, a fully grown amphipod would be way too big to put in an electron microscope. A regular stereo microscope would be sufficient for imaging that. Thank you for the ostrscod idea. After some research, that was my thought as well.

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Thank you amphipod! Believe it or not, a fully grown amphipod would be way too big to put in an electron microscope. A regular stereo microscope would be sufficient for imaging that. Thank you for the ostrscod idea. After some research, that was my thought as well.

what's the maximum diamater in millimeters of an object to fit in? Something cool to see under a microscope would be a Microcerberidea isopod, but I don't think you'll be catching one anytime soon.
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HarryPotter

Cool! Amazing images- never seen something like them outside my

Textbooks!

 

Okay, a few weird things I'm interested in seeing up close: a dirty penny, Dino vs GHA, a clam's mantle, the follicles of GSP, ICH'

 

ICH! Can you 'scope that? It's bacterial,

Right?

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Mr. Microscope

what's the maximum diamater in millimeters of an object to fit in? Something cool to see under a microscope would be a Microcerberidea isopod, but I don't think you'll be catching one anytime soon.

You can fit fairly large items into some SEM's. We've even had ancient jade pots (from the museum) in one of ours, but the minimum magnification is quite high. So, if you put a large object into an electron microscope, it's like trying to understand a soccer game by looking at a blade of grass on the field.

 

 

Cool! Amazing images- never seen something like them outside my

Textbooks!

 

Okay, a few weird things I'm interested in seeing up close: a dirty penny, Dino vs GHA, a clam's mantle, the follicles of GSP, ICH'

 

ICH! Can you 'scope that? It's bacterial,

Right?

Thank you Harry! For starters, I will be posting some dinoflagelate images later today or tomorrow. So, you'll get to see some of those soon. I assure you, they look nothing like GHA.

EDIT: I found this video I made back in March of 2014. Near the end, you can see both dinos and GHA.

Annette, these aren't yours.

 

A clam's mantle is a bit large for EM, but I suppose if I could get my hand on some and cut it up, I might have something cool to look at. GSP would probably be tough to keep happy and image, but if I fixed it with some aldehydes, it might maintain its shape. Ich is a definitely possibility. Though, it would need to be propagated first.

As for a dirty penny, I actually looked at one when Keyence came to demo their microscope. I don't normally post non-reef related images here, but by request here is what I believe to be a oxidation pit on a penny.

VHX_000048_zpssvp5xcae.jpg

Also, did you know there is a second Lincoln on the penny? It's in the memorial on the back. Utilizing the Keyence focal stack, you can really see it illustrated.

penny-backside-50x_zpslzuxuslm.jpg

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charnelhouse

You can fit fairly large items into some SEM's. We've even had ancient jade pots (from the museum) in one of ours, but the minimum magnification is quite high. So, if you put a large object into an electron microscope, it's like trying to understand a soccer game by looking at a blade of grass on the field.

 

 

Thank you Harry! For starters, I will be posting some dinoflagelate images later today or tomorrow. So, you'll get to see some of those soon. I assure you, they look nothing like GHA. A clam's mantle is a bit large for EM, but I suppose if I could get my hand on some and cut it up, I might have something cool to look at. GSP would probably be tough to keep happy and image, but if I fixed it with some aldehydes, it might maintain its shape. Ich is a definitely possibility. Though, it would need to be propagated first.

As for a dirty penny, I actually looked at one when Keyence came to demo their microscope. I don't normally post non-reef related images here, but by request here is what I believe to be a oxidation pit on a penny.

VHX_000048_zpssvp5xcae.jpg

Also, did you know there is a second Lincoln on the penny? It's in the memorial on the back. Utilizing the Keyence focal stack, you can really see it illustrated.

penny-backside-50x_zpslzuxuslm.jpg

 

I knew that! You can kind of see it with the nekkid eye too...

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