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Calpain-EqD — DIC and Antibody Stains
Published: 2012-05-26 20:35:49 +0000 UTC; Views: 1850; Favourites: 15; Downloads: 16
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Description
This pic ended up being one of the figures in my paper that is off for review at the moment. While I didn't get to finish the experiment these photos are associated with, I was proud of being able to optimize the procedure to a point where if someone else were to pick up the project they should have an easier time completing it.What you see here are a DIC picture and three antibody pics on fly neurons I grew in culture months ago. The DIC picture in the upper left was taken to just see if the cell population was healthy, as distinguished by the appearance of processes extending from the neurons and by the degree of cell spread.
The antibody pics are of: actin in the upper right corner (actin is used in cells for the construction of microfilaments, a component of the cytoskeleton in cells), beta-tubulin in the bottom left (tubulin is used in constructing microtubules, a component of the cytoskeleton and the cell's transportation network) and finally tau protein in the bottom right (tau is a microtubule assisting protein which helps microtubules assemble and disassemble. They are also suspected agents in Alzheimer's disease).
Anyhow, I was rather happy with this plate so I thought I'd share! Hopefully you learned a little while you were here!
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Comments: 20
First thing that came to my mind when I saw DIC was disseminated intravascukar coagulation XD darn. Nice photos by the way! Only if we could reverse the accumulation of those proteins somehow (or prevent accumulation in the first place). Go science!
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Nice! right now i am modding a second hand microscope i got, to be able to do fluorescence microscopy, but i will be stuck to UV fluorophores for some time, because i am using a nitrogen Laser as light source (337.4nm) which is a wavelength easy to block with a polycarbonate filter without interfering with the florescence light of the specimen.
While i am thinking of pumping a fiber coupled dye laser to be able to use almost any fluorophore the dichroic filters to block the reflected excitation light are hard to find and expensive 
On another note, have you used Super-resolution microscopy to image the tau protein working inside the cell? i don't know how easy or even if you can tag τ proteins to be able to observe them (i am just starting to learn about all this) but i would like to see a time lapse of cells growing and do a comparation between a healty cell and one with Alzheimer's disease. I wish you good luck with your Science!
Scientia et Equus
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This is pretty cool
I love when science alone can look like art
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Can you link the paper in any way, shape or form? I'd be interested to read it!
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Very interesting... Do go on!
We had to do micro shots for metallurgy class in engineering, and this picture reminded me of it.
Here's a link with description, don't want to thread crap with a huge picture: [link]
In my shot I see some of the main details such as the carbide balls that have floated to the top, and some of the micro-cracks going down into the structure.
What you have in your pictures though... Those are living things! And, well, that is just fascinating that they are that small, and you are just scratching at the surface so to say!
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That b-tubulin stain turned out absolutely beautiful! And the tau/actin stains actually match up quite nicely (as I suppose they should). You mentioned Alzheimer's. Was this part of a procedure to test neurotoxins against the survivability of neurons?
I would guess this -might- not be the results (just the procedure) cause the neurons look pretty healthy to me.
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Thanks for the compliment! It was actually part of a procedure to see if tau protein colocalized with the protein cutter known as calpain in neurons. It was a work in progress and we never reached the final steps, but the procedure on how to grow neurons was finalized with these experiments so we at least got something out of it.
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wait, so your nickname is based off of a protein cutter?!?!?! that's... brilliant!!! o,0
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Yay, now I feel like a smarty pants because I was able to retain information from my ninth grade biology class and therefore comprehend what you were saying.
Now I also kind of regret not taking AP Biology.
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Biology classes have always been a joy for me! If you can, try and take another sometime I don't think you'll regret it.
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lol idk any bout DIC or biology stuff but it looks great! I think... <3
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Heya Solar! DIC is actually a method we use to see transparent or nearly transparent organisms/tissues/ect. By skewing the light just slightly as it passes through the object, you get two offset images. When the scope puts them back together again, you get a shadow-like or 3D effect that makes invisible objects stand out and become visible! It's pretty cool.
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Sounds so scientificial and stuff lol
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science is magic.... wait, i think i just broke too many laws with that sentence u,u
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Oh wow.
Most of this stuff does go over my head, but it's really interesting to see these and their explanations.
Keep up the good work!
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Thanks! This sort of stuff is pretty fun to do. It's amazing when an experiment comes together.
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