MicrobeHunter.com Microscopy Forum

Someone from Nikon stopped by to help us install an incubation unit on our confocal scope. Someone from a different lab brought a GFP tagged tubulin cell line with him to come give it a test and we had a look. I watched tubulin moving around in a cell in real time at a very fast frame rate in 3D. Ne...

There's one particular algae I have that doesn't take so well to formaldehyde, glutaraldehyde, or any other fixing agent I've tried. They usually just explode when I add anything. So far I've been able to stun them with nickel sulfate but that irritates them a bit too. Has anybody tried just throwin...

Thanks for the reply. It is a fairly high end camera. I thought this would be more like fitting a low end lens to my camera, which I've done before. That's pretty much what's going to happen, the microscope is going to be your limiting factor here. If you want to do some microscopy and don't want t...

Not that you'll find this on any of the equipment for home use, but last high end Leica I used had reaction wheels on the parts that moved the stage to soak up movement from the stage moving around. Haven't noticed if they're on the confocal I use. The Leica ones were bright yellow, maybe they're ju...

All the microscopes I use are on air tables and really heavy and I still step refrain from moving or touching anything when I take photos. If you don't have one, you should definitely get a cable release for the camera. Also always use mirror lockup.

So I should expect a trained ciliate army ready to take over the world soon then?

I think what people mean is don't try to self diagnose using your own microscope, although I don't think you were trying to do that. In reality it's not like they'd diagnose you of any kind of bacterial infection with a microscope. I wouldn't even do it with different agar's, I'd just throw them on ...

When I gram stain, I put some water on the slide, spread it out, take a loop or some other instrument and mush it around in a colony, then place the loop in the water and move it around. Then I heat fix to the slide by running it over a flame for a bit just so the water goes away. Then you hit it wi...

You mean to seal the coverslip to the slide or as the actual mounting medium? I've used it to seal coverslips until I started using som hard setting mount.

Kinase, That should work in theory, but treating a plant living in soil is pharmacologically different than treating cultured cells (lots of colchicine may be needed). It may be easier for a person to culture freshly isolated plant tissues (in some culture fluids) for a few days with the right amou...

Looks like one chromatid has it's top part in what is going to be one nucleus and its bottom in whats going to be the other nucleus. I've had plenty of lectures on mitosis and there's some really cool TEM's and stuff about how exactly the machinery works. I wonder what caused that exactly though. I ...

For all the fancy kinds of staining methods I use, I pretty much never use regular non-fluorescent stains. These come out really nice, great photos.

What exactly is it you're trying to figure out? The confocal is also capable of epifluorescence, at least the one I use also has an LED source for 405, 488, 561, 640. That link goes to vasopression with BIotin tagged to it, are you going to use an anti-biotin antibody and then secondaries to see whe...

DNA strongly absorbs UV light, 260nm is the peak. I think that laser was a 355nm but it works just fine. Some people went through the trouble of validating the method awhile ago and went about proving that this is actually laser induced DNA damage. The idea is to selectively damage a region of the n...

good stuff! 2nd picture is confocal? Agree keep it up! The Nikon scope with the stripe is not confocal, that box thing on the left is the IR/UV laser unit. That Eclipse Ti is the inverted base for the confocal I use though. Here's one off the confocal. This was on a Nikon CFI Plan Apochromat λ 60X ...

The last of the demo scopes. This is a Nikon Eclipse Ti with an Applied Biosystems laser unit. This one has a UV laser for cutting and an IR laser for laser tweezers. I didn't make use of the IR laser but some other people were doing actual microdissection and cutting out pancreatic islets from a ti...

The lab who has that microscope does this all the time, like their whole lab depends on the ability to cause DNA damage like that. Their old machine sort of works but they're already cleared to get a new one so they're going to. I'm not sure what happened but I'll figure it out when I get back from ...

Somehow I ended up doing no apparent DNA damage to the cells, not even near where I etched the glass... Something's wrong here but I'm not sure what yet... The only thing I could think of is that the glass doesn't transmit the wavelength of laser we had, but the media boiled in some cases when I fir...

Even us "professionals" like to have some fun with the equipment. Another lab's laser microdisection unit was broken so they're demoing new ones. Personally I liked the Leica unit they demo'd more than any of the other ones. At any rate, a Zeiss inverted scope with a 3rd part laser unit is here. I t...

Yes, thank you kinase. Keep these post going please. Its really nice to see people working with professional kit. Really interesting to get a window on professional work. Many thanks billben Ha I'm not sure if professional is the right word yet, I'm still a student, hoping to be a PhD student next ...

For this photo, I seeded cells on a #1.5 coverslip that I sterilized in 100% EtOH. I treat the cells with IDU (iododeoxyuridine) as a radiosensitization agent because it incorporates into DNA but doesn't quite fit very well in the helix, creating a fragile site that will easily be damaged upon appli...

https://www.fbi.gov/about-us/lab/forensic-science-communications/fsc/july2004/research/2004_03_research02.htm I just barely scanned part of this, it looks fascinating. Now you've got me interested! Dale I was going to say, I'm pretty sure they use transmission or scanning electron microscopes for f...

You might think tissue culture requires intense precautions for sterility but it doesn't. A pair of gloves, sterile media, generous amounts of 70% EtOH, a biosafety cabinet, an incubator, and some common sense are all you need really. And sterile culture dishes. Plus the media has penicillin/strepto...

Microscope porn thread. There's a few I don't have pictures of, namely the Zeiss axiovert 100M I usually use and a Nikon A1R+ I recently started using, and the microscopes in cell culture room which aren't anything to write home about. http://i1219.photobucket.com/albums/dd424/lolmywallets/IMG_6605_...

I'm using it as a method to say that two proteins are close enough to potentially be interacting with each other, i.e. within a couple nanometers of each. It's a way to get around the diffraction limit, because I know certain properties of the fluorophores I'm using I can tell if they're close to ea...

I'm certain this a long shot but has anybody done FRET before? I'm planning a acceptor photobleach FRET experiment and I'm pretty sure I have everything in working order. I was just wondering if anybody had experience with it and might be able to offer insight on what the best way to do the actual p...

It's definitely a lot of practice. I'm pretty sure someone at Zeiss invented and patented DIC and hence the big four have different ways of doing it. I really like DIC. Once you get good at it you can do a lot of stuff. I was even able to get fluorescence and DIC working at the same time in one case...

Nikon's page as well a Zeiss and Olympus and probably Leica have a bunch of pages on information about objectives and everything else they sell. I wish I had a 63x or thereabouts oil lens. On the scope I usually use, a older Zeiss axiovert 100M I have a 40x and 100x oil that I like. On an Olympus I...

If I can, I try only to use oil lenses for taking pictures. I only use air lenses for checking confluence on plates really. I haven't had a chance to use it yet because I've been unable to stain my samples but I assume the Nikon A1R the facility I'm in has uses fancy glass. The microscopes in cell c...

I just had a thought, and it may be a stupid one but here it goes - is there anything stopping me from taking an upright, turning it upside down, and using it as an inverted? Other than the inconvenience of everything now being upside down... I don't really see why it wouldn't work? I would keep the...