Want to See Something Microscopic in 3D?

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Circlenaut

Platinum Member
Mar 22, 2001
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:thumbsup:

they have really cool heads.
:D

Hmm, this got me thinking. Maybe I could take a slice of where the optic nerve meets the visera so I could get a "topography" of the surface of rods and cones. The detection molecules have to contain chromophores, I'll check out what their excitation wavelengths are.

On another note I thought about using a cpu die but because it's flat I don't think it will be much different than what a regular scope would be. So far I'll be looking at diatoms (these could turn out very pretty), fly heads, rods and cones in eyes and mushroom spores.
 
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Anonemous

Diamond Member
May 19, 2003
7,361
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For the confocal microscopes I've used, the laser(usually) has to be able to pass through the sample. It also needs to be a thin sample (100-200um thick depending on the working distance and magnification).
 

Circlenaut

Platinum Member
Mar 22, 2001
2,175
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100 million per load... there is no such thing as overkill when it comes to semen.

Hmm I could check this out with a phase contrast scope first. What restaurant do you have in mind, If it's in the Worcester, MA area I could pick one up to test out.
 

Circlenaut

Platinum Member
Mar 22, 2001
2,175
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81
For the confocal microscopes I've used, the laser(usually) has to be able to pass through the sample. It also needs to be a thin sample (100-200um thick depending on the working distance and magnification).

Yeah you're right, the light needs to pass through.
 

Cerpin Taxt

Lifer
Feb 23, 2005
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For the confocal microscopes I've used, the laser(usually) has to be able to pass through the sample. It also needs to be a thin sample (100-200um thick depending on the working distance and magnification).

This is not really true. As far as I know, all laser scanning confocal microscopes capture reflected light, although in multiphoton confocal microscopy the imaging plane is generally quite deep below the surface of the tissue (500um to 1000um, IIRC). I am not aware of any strict requirement for the sample to be able to transmit the laser through it.

Likewise, there is no strict limit to the thickness of a sample, however there will be a limit to how deep within the tissue the imaging plane can be, depending on the type of laser excitation you're using.

But what do I know. ;)
 

Circlenaut

Platinum Member
Mar 22, 2001
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This is not really true. As far as I know, all laser scanning confocal microscopes capture reflected light, although in multiphoton confocal microscopy the imaging plane is generally quite deep below the surface of the tissue (500um to 1000um, IIRC). I am not aware of any strict requirement for the sample to be able to transmit the laser through it.

Likewise, there is no strict limit to the thickness of a sample, however there will be a limit to how deep within the tissue the imaging plane can be, depending on the type of laser excitation you're using.

But what do I know. ;)

From what I understand how our confocal scope works it seems that the laser comes in from the bottom of the sample, hits it and causes any dye/florescent protein to glow (if excited in the appropriate wavelength). So I imagine that the sample has to be thin enough for the laser to excite the molecules and then thin enough for the excited light to exit.
 

Cerpin Taxt

Lifer
Feb 23, 2005
11,940
542
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From what I understand how our confocal scope works it seems that the laser comes in from the bottom of the sample, hits it and causes any dye/florescent protein to glow (if excited in the appropriate wavelength). So I imagine that the sample has to be thin enough for the laser to excite the molecules and then thin enough for the excited light to exit.
Unless your set up is something relatively unusual, I'm fairly confident the emitted light exits the same way the excitation light came in.

Where is the camera? Is it on the same side of the sample as the laser? What microscope frame are you using? Zeiss? Leica? Olympus? It is an inverted microscope, yes?
 

zinfamous

No Lifer
Jul 12, 2006
111,858
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On another note I thought about using a cpu die but because it's flat I don't think it will be much different then what a regular scope would be. So far I'll be looking at diatoms (these could turn out very pretty), fly heads, rods and cones in eyes and mushroom spores.


THAN




(caps)
:sneaky:
 

Mr. Pedantic

Diamond Member
Feb 14, 2010
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Surely taking a confocal of a CPU die won't work, because the structure in the die are too small for light...?
 

Circlenaut

Platinum Member
Mar 22, 2001
2,175
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Unless your set up is something relatively unusual, I'm fairly confident the emitted light exits the same way the excitation light came in.

Where is the camera? Is it on the same side of the sample as the laser? What microscope frame are you using? Zeiss? Leica? Olympus? It is an inverted microscope, yes?

It's http://www.leica-microsystems.com/products/confocal-microscopes/details/product/leica-tcs-sp5-ii/ this guy but number I. The professor also said they may have added a UV laser this year.
 

fralexandr

Platinum Member
Apr 26, 2007
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Quote:
Originally Posted by Pippy View Post
Hmm, I know one of the Professors in the Dept. works with Drosophilae. I'll see about getting a head and mounting it. I'll definitely be cool to see the eyes.


they have really cool heads.
__________________

oo how about one of the mutants with arms for mandibles

also how does one nested quote things? >.<
 
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Circlenaut

Platinum Member
Mar 22, 2001
2,175
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81
Surely taking a confocal of a CPU die won't work, because the structure in the die are too small for light...?

Heh you're right I should have thought about this last year when I took TEM. 45nm is way past the resolution of the best optical microscopes (~200nm)
 

Mr. Pedantic

Diamond Member
Feb 14, 2010
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If you can change the frequency of the light though, high-energy UV will still do 45nm, but if someone has a Pentium Pro or something normal blue light should do the trick.
 

zinfamous

No Lifer
Jul 12, 2006
111,858
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oo how about one of the mutants with arms for mandibles

also how does one nested quote things? >.<

ah, you mean antenaepoidia? (legs growing out of antennae stalks)


eh, interesting freak critter for sure, but might as well just look at a leg or an antennae, though.
 

DrPizza

Administrator Elite Member Goat Whisperer
Mar 5, 2001
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www.slatebrookfarm.com
A fly would be pretty cool.

Or, pull some eyelashes out. If I recall correctly, most adults harbor some sort of parasite that lives in the hair follicles. It'd be pretty cool to see those parasites in 3-D.