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What microscope for movies like these? - ( new member)

 
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Thomas Ashcraft



Joined: 31 May 2006
Posts: 25

PostPosted: Fri Jun 02, 2006 3:36 pm    Post subject: What microscope for movies like these? - ( new member) Reply with quote

Greetings to all of you who think small like I do. My name is Thomas Ashcraft and I am a new member living in New Mexico, USA. I am in the process of manifesting the right microscope for videos and photography and could use a little advice as I am in the steep part of the learning curve at present and trying to absorb as much information as possible.

I discovered some dynamic short bacteria movies on the web and would like to ask a question as what sort of microscope would be capable of this sort of bacteriological subject:

The movies of interest are at: http://cmgm.stanford.edu/theriot/movies.htm#Hits

Specific examples on that page are:
"Listeria "hopping" in Xenopus extract"
"Listeria monocytogenes travelling in spirals in Xenopus extract"

Would a basic phase contrast light microscope at 950 or 1000x be capable of this sort of movie? ( And with resourcefulness, would a two thousand dollar budget be adequate to purchase a quality microscope and get going?)

Thank you for any replies!

Thomas
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Charles Krebs



Joined: 13 Jul 2004
Posts: 1200
Location: Issaquah, WA USA

PostPosted: Sat Jun 03, 2006 10:49 am    Post subject: Reply with quote

Greetings Thomas

You are aiming high Wink . Since the work you are referencing is being done at Stanford, you can imagine that they are working with some of the the best equipment available. If you look at the research papers they have posted (in PDF format) some of them give a brief description of what is used. For example:


http://cmgm.stanford.edu/theriot/pdf%20files%20of%20papers/Cameron%20etal2004.pdf :
Quote:
Most observations were performed on an Axioplan2 microscope (Carl Zeiss, Thornwood, NY) equipped with phase contrast and epifluorescence optics. Time-lapse microscopy was achieved with a digital cooled charge-coupled device (CCD) camera (Micro-MAX:512BFT, Princeton Instruments, Roper Scientific, Trenton, NJ) specialized for low light level and high frequency imaging, by using MetaMorph (Universal Imaging, Media, PA) software. Every 10 s, phase contrast (with 50-ms exposure) and fluorescence (400-ms exposures) image pairs were recorded.
Some observations were performed on a Nikon Diaphot-300 inverted microscope equipped with phase contrast and epifluorescence optics with an attached intensified CCD camera (Dage-MTI GenIISys/CCD-c72) for timelapse videomicroscopy. Video images were captured, digitized, and analyzed using MetaMorph software. Phase contrast and fluorescence image pairs were recorded every 10 s with eight video frames averaged at each time point.

Another document listed the same microscopes and added the objectives used: 63/1.40 and 100/1.40 on the Zeiss; 60/1.40 on the Nikon. The very high numerical apertures (1.40) give the highest possible resolution with a light microscope... essential for studying bacteria.

Fluorescence in this "league" will be tough with your budget, but you should be able to get set up with phase-contrast. You will want a trinocular head for the photography. I don't know much about video and time-lapse video, so hopefully others will provide some info.
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Thomas Ashcraft



Joined: 31 May 2006
Posts: 25

PostPosted: Sat Jun 03, 2006 7:32 pm    Post subject: Reply with quote

Hi Charles,

Thanks for that detailed reply and for generously digging out those references. I have spent the time since your reply at "Nikon Microscopy U" on the web learning as much as possible. ( That's a pretty good resource!)

Indeed, Stanford has enviable resources for sure. Yet it's also nice to have a personal lab and own your own time and data and practice old fashioned independent naturalism. Smile

Interestingly, as I started googling epi-fluorescence possibilities it appears that a Nikon E200 microscope has fluorescence add-on capabilities ( though I never found a posted price for this particular microscope anywhere. ) With further googling I came across one of those questionable and inexpensive Chinese Ebay microscopes that is a full epi-fluorescence system. I would not trust the mercury elements or other tight tolerances in a supercheap scope like that though. But it is interesting that it is offered in any case.

Many thanks again and hopefully I will be set up for phase contrast soon. Until then, I will be studying things like cover slip thicknesses and immersion oil details at "Nikon Microscopy U."

Good observing to you.

Thomas
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wilash



Joined: 13 Apr 2005
Posts: 104
Location: Japan

PostPosted: Sat Jun 03, 2006 9:33 pm    Post subject: Reply with quote

If you don't mind used equipment, you can get some great deals on ebay - go to the business and industrial section and seach for a manufacturer's name such as Olympus, Nikon, Zeiss, Leica, Wild, American Optical, etc. Then start digging. I bought an Olympus Vanox research microscope with DIC for a fraction of the cost. But do reseach on microscopes first. There are scopes from many different vintages on ebay and you need to realize the differences in optics. My scope was designed before infinity optics, but that was OK for me as I figured the optics would still could give me good results at least equal to if not better than an inexpensive low-end model AND it was a tad cheaper. Also I watched the actions long enough to realize that objective lenses came up often enough and at good prices not to worry about the supply.

If going secondhand, I would recommend getting the scope already fitted out with what you want. In my case I was looking for an episcopic scope with DIC and a large-format camera. Depending on the age of the scope, it may be tricky picking up the parts you need. I did upgrade my camera later with an electronic camera to replace the mechanical shutter.
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Ken Ramos
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Joined: 04 May 2004
Posts: 4809
Location: Western North Carolina

PostPosted: Sun Jun 04, 2006 3:43 am    Post subject: Reply with quote

For what it may be worth. I noticed an Axioplan in the equipment line up. Most of you know that I have an Axiostar and here a while back I was thinking of upgrading to an Axiophot. That microscope alone with only the basics is around $4000.00 or better! Rolling Eyes So I would imagine the Axioplan is much more, not counting the high NA objectives and other bells and whistles required. Wink
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Kens Microscopy
Reposts of my images within the galleries are welcome, as are constructive critical critiques.
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Thomas Ashcraft



Joined: 31 May 2006
Posts: 25

PostPosted: Wed Jun 07, 2006 5:55 am    Post subject: Reply with quote

"If you don't mind used equipment, you can get some great deals on ebay - go to the business and industrial section and seach for a manufacturer's name such as Olympus, Nikon, Zeiss, Leica, Wild, American Optical, etc. Then start digging. I bought an Olympus Vanox research microscope with DIC for a fraction of the cost. "

Hi Will, glad you got a great deal on eBay, especially for the DIC feature! And thanks for your reply.

I have been studying eBay but see that getting a truly sweet deal will require a lot of patience. It would be nice to put together an old Zeiss or Ortholux from parts though. I am glad there is an ebay.

Thomas
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Thomas Ashcraft



Joined: 31 May 2006
Posts: 25

PostPosted: Wed Jun 07, 2006 6:45 am    Post subject: Reply with quote

Ken wrote: "...... I was thinking of upgrading to an Axiophot. That microscope alone with only the basics is around $4000.00 or better! Rolling Eyes So I would imagine the Axioplan is much more, not counting the high NA objectives and other bells and whistles required."

Hi Ken,

That's serious money! My ponderence now is whether a top of the line Meiji or Motic with plan objectives can approach a Zeiss like you are mentioning in image quality and performance for a fraction of the cost.

It is challenging indeed to shop for a microscope.

Good observations to you,
Thomas
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