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Microscope specs

 
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CaptainNemo



Joined: 28 Nov 2016
Posts: 2

PostPosted: Sun Dec 04, 2016 9:37 am    Post subject: Microscope specs Reply with quote

Hi all,

I am a single molecule detection microscopist and interested in starting photomicrography. As you all are very experienced:

What microscope specs should I be looking for! Trinocular ok....but then what? And where can I look for them?
Suggestions?

Thanks and looking forward to spending some time here.
Captain Nemo
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JohnyM



Joined: 24 Dec 2013
Posts: 166

PostPosted: Sun Dec 04, 2016 10:14 am    Post subject: Reply with quote

Single molecule detection microscopy systems are working with PMT systems, and dont require trinocular tubes, as PMT are located inside scan head. Afaik only TIRF systems use CCD cameras, and im not sure if nipkov type is suitable for single molecule microscopy.
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CaptainNemo



Joined: 28 Nov 2016
Posts: 2

PostPosted: Sun Dec 04, 2016 10:24 am    Post subject: Whoops Reply with quote

Oh, some confusion here, i am not interested in doing single molecule detection, i was merely talking about my background. I would be interested in going to the cellular level, looking at cells, looking at bacteria, diatomees, foraminiphera, ... Single molecule fluoescence microscopy and spectroscopy is what I did in the past during my phd.
And there we used single photon detectors and nitrogen cooled CCD cameras.
Best
Captain nemo
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JohnyM



Joined: 24 Dec 2013
Posts: 166

PostPosted: Sun Dec 04, 2016 11:42 am    Post subject: Reply with quote

So you already got top class equipment, eazy to adapt it to widefield microphotography. Give me a name or photo of your microscope, so i could advize you what you need.
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zzffnn



Joined: 22 May 2014
Posts: 1487
Location: Texas USA

PostPosted: Sun Dec 04, 2016 12:00 pm    Post subject: Reply with quote

Welcome to the forum.

Please tell us your budget.

Look into DIC, darkfield, phase contrast and oblique illumination. And figure out which you like to have and can afford. Having a PhD in the field, it should be easy for you to figure it out yourself.

If you have over $2000 in budget, I suggest going with DIC. If not, try to get good apochromatic objectives for your most useful magnification (which you need to figure out yourself too, for subjects you mentioned, it is mostly like equal or more than a 40x objective).

When I started, my budget was $500-$1000, though I ended up spending more. I personally like pond protists and do lots of live water mount, so I stay with oblique, darkfield and water immersion LOMO objectives. I cannot afford DIC and personally do not like phase contrast. I stay with 160 mm tube length system due to better optical compatibility between brands, which makes it cheaper to get features I want too.

Your imaging rig may depend on your camera / sensor size and lens / objectives combinations too.

Having an aplanatic achromat c condenser helps too, if you can afford it.

So it depends on what you have and what you want.

Edit:

Some of the most important specs in microscopy are, but not limited to:

NA, which is related to resolution. Technically the higher the better, but higher resolution typically correlates to less working distance and more difficulties in sample preparation and illumination, especially at over NA 0.9.

Magnification. It will be empty magnification (without increased resolution), once it goes over 1300x. Also focal length.

Tube length. 160 mm or infinity.

Field number (width of well-corrected view field). Your eyes will appreciate at least 18mm of such.


Last edited by zzffnn on Sun Dec 04, 2016 12:57 pm; edited 6 times in total
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Pau
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Joined: 20 Jan 2010
Posts: 4003
Location: Valencia, Spain

PostPosted: Sun Dec 04, 2016 12:32 pm    Post subject: Reply with quote

Budget?
This is a first relevant question, of course not the only relevant one.
Illumination techniques you want to do?
New or used? (new good equipment is so expensive that is prohibitive for most people for personal use). There are new low priced instruments but not very capable...

My approach was old stuff (Zeiss Standard) from the 60s-80s bought almost piece to piece and with some DIY adaptations. It ended (after around 4 years ...) being a very capable and versatile system for a moderate budget. Undoubtedly there are more judicious options
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Perl



Joined: 03 Oct 2006
Posts: 206
Location: Sweden

PostPosted: Mon Dec 05, 2016 12:55 am    Post subject: Reply with quote

Hi
Do as Pau - but switch to Leitz after a while
due to Optical separations problem in Zeiss
microscope optics from that era

Now ended up ( 10 years after ) with anything made for
Leitz Microscope in 70-80 and it take space - but absolutley fun

Regards
Pär
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