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PostPosted: Tue Apr 16, 2019 1:36 pm 

Joined: Thu Nov 29, 2018 12:59 pm
Posts: 4
I recently procured an entry-level fluorescence microscope, am now considering camera system for that. I’m using the microscope for scientific research.

I’m not sure if it is worthy to buy specialized microscope cameras. Pre-owned older models of these cameras are more expensive than pre-owned full frame DSLRs. Once I obtain these used specialized cameras, I also need to put some efforts to make them work; e.g. to find obsolete desktop with WIN XP installed to accommodate the PCI interface board…

I think low light performance is important for fluorescence microscopy. The specialized camera has pixel size 6-7 um, bigger than that of FF DSLR sensors and much bigger than that of APS-C sensors. Also, specialized camera CCD is cooled during operations. I think these features benefit the low light performance of specialized cameras. However, DSLR has the advantage of larger sensor and more pixels. DSLR can work at 6400 or even 12800 ISO. But some older specialized microscope cameras work has ISO level no more than 1600. Overall, which has better low light performance?

Specialized cameras are tailored for microscopes, especially for microscopes from the same manufacturer. They can be controlled by professional software for shooting parameters. Meanwhile, I don’t know how can I control the DSLR attached to a microscope, which mode should I shoot with?

So, please kindly advise, which camera system should I take, any issues omitted from my consideration?

PostPosted: Tue Apr 16, 2019 3:09 pm 

Joined: Mon Aug 21, 2017 9:02 pm
Posts: 1690
It is difficult to answer with certainty about which camera you need, especially since you provide no details about your microscope, your future specimens and what experiments shall be done. So I will try some general suggestions.

As you probably know, fluorescence units are arbitrary, not absolute. Fluorescence intensity depends on several factors: chiefly on the quantum yield of the observed specimen and the intensity of the excitation - in addition to the obvious dependence on wavelength. The microscope optics, including the objectives, dichroic mirror and excitation and emission filters also play a role. Often one needs to work in a dark environment to avoid stray light. So a given camera might be suitable or not.

I think that low light level performance (sensitivity, signal-to-noise) of the camera is crucial, and is more important than pixel resolution. Full control of the camera by means of the computer software is, I think, also very important for research. A dedicated computer card like the PCI you mentioned, is IMO only necessary if you plan a very high rate of transfer of high-resolution images to the computer. Otherwise, the dedicated card on obsolete computer motherboard is not critical.

I am guessing that you are starting a research on a budget. Top level fluorescence research data rely on dedicated microscope cameras - say, Olympus DP-74 (just an example) or, for confocal microscopy, photomultipliers. The latter are much more sensitive and fast and linear than cameras (a generalization, not overwhelming). And the microscope manufacturer's software. I would choose such camera rather than a DSLR.

One other point that you might want to consider is the coverage of the field of view. For example, the Olympus DP-X (X=70, 73, etc) camera, on top of a BX-series microscope is a wonderful modern setup. Yet, FOV coverage is far from complete, unless you specifically add a relay 0.3X or 0.5X lens above the objective.

May I suggest the following approach: prepare some test slides. The easiest, and most reproducible, are fluorescent micro- or nano- spheres. They are made of latex or polystyrene or other polymer, at sizes of 100nm-10micron (at least), and are stained with a monochromatic or polychromatic stains. They give off a very bright fluorescence that is quite immune to photo-bleaching. They are uniform in size. Can be mounted dry, or in a liquid or resin medium. They are available from most chemical/biological supply houses. Some photos of such slides, perhaps with a (loaned) camera, will provide a qualitative and quantitative estimate of the performance of the camera.

Zeiss Standard GFL+Canon EOS-M10, Olympus VMZ stereo

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