For fluorescence microscopy, specialized microscope camera or DSLR?

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tocharian
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For fluorescence microscopy, specialized microscope camera or DSLR?

#1 Post by tocharian » Tue Apr 16, 2019 1:36 pm

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?

Hobbyst46
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Re: For fluorescence microscopy, specialized microscope camera or DSLR?

#2 Post by Hobbyst46 » Tue Apr 16, 2019 3:09 pm

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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RobBerdan
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Re: For fluorescence microscopy, specialized microscope camera or DSLR?

#3 Post by RobBerdan » Sun Aug 09, 2020 4:12 am

I realize this post is over a year old, but I feel compelled to answer it. I own a Fluorescence microscope Nikon Optiphot as well several other scopes. I have been testing different models of dedicated cameras including Zeiss, AM systems, Moticom and Rising Cam. The Rising cam offers 20 MP and is the best dedicated camera I have tested so far - its available on Ebay for about $800.

The other camera cost more and preformed more poorly. I have also been using a variety of DSLR cameras including both Nikon and Canon for many years and can say they perform much better then any dedicated camera so far. They are excellent for Fluorescence, you can use free software Digicam control to view specimens in Live view and take both pictures and movies. I took a movie of paramecium at 30 fps after staining with Acridine orange. You can increase the ISO speed usually 400-1600 is good. Having good fluorescent objectives with high NA also helps. Shoot RAW files and process in Photoshop.

I will be posting an article in a few weeks which compares DSLRs vs dedicated cameras and will even results with Cell phones.
To see sample images in Fluorescence taken with a DSLR see:
https://www.canadiannaturephotographer. ... scopy.html
https://www.canadiannaturephotographer. ... fers2.html

I also describe how to use Photoshop to apply Deconvolution to the images.
I will show in the near future the how DSLRs are supperior to dedicated cameras in most circumstances.
R.B
Attachments
Brachionus rotifer stained with Acridine orange - Nikon D500 DSLR
Brachionus rotifer stained with Acridine orange - Nikon D500 DSLR
DSC_0270.jpg (57.33 KiB) Viewed 1304 times

Hobbyst46
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Re: For fluorescence microscopy, specialized microscope camera or DSLR?

#4 Post by Hobbyst46 » Sun Aug 09, 2020 5:56 am

RobBerdan wrote:
Sun Aug 09, 2020 4:12 am
...I will show in the near future the how DSLRs are supperior to dedicated cameras in most circumstances.
Looking forward to see it !
The factors of sensor size (much smaller in a dedicated camera than in the DSLR), low-light sensitivity (higher in dedicated cameras) and data transfer speed come to mind.
Hopefully you included mirrorless cameras and not only DSLRs.
Zeiss Standard GFL+Canon EOS-M10, Olympus VMZ stereo

MichaelG.
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Re: For fluorescence microscopy, specialized microscope camera or DSLR?

#5 Post by MichaelG. » Sun Aug 09, 2020 6:53 am

Hobbyst46 wrote:
Sun Aug 09, 2020 5:56 am
RobBerdan wrote:
Sun Aug 09, 2020 4:12 am
...I will show in the near future the how DSLRs are supperior to dedicated cameras in most circumstances.
Looking forward to see it !
Me too !
Your practical experience is enormously valuable, as your results demonstrate.

MichaelG.

.

But, just for balance, I will mention this ‘case for the defence’ presented last year:
viewtopic.php?f=9&t=7728&hilit=keal
Too many 'projects'

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hkv
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Re: For fluorescence microscopy, specialized microscope camera or DSLR?

#6 Post by hkv » Sun Aug 09, 2020 11:05 am

I totally agree with Robert here. I have also tested many microscope cameras, but I produce better images with DSLRs. One of the biggest issues I have had over the years with dedicated microscope cameras is that they require proprietary software which often is buggy and slow. Commercial software and even free software for DSLR are normally more mature and issues are being fixed quickly. Also, the user community is much larger which means that whatever questions or issue you may have, there is always someone that has experienced it before and the solution is almost always available online in some forum.

When it comes to image quality I see no drawback what so ever using DSLRs except for specific applications where you need extremely long exposure times, like more than 30 seconds and up. Then a cooled sensor may be an advantage that some of the more expensive dedicated cameras may have. I have never seen the need for that in my work as a normal "micro photographer".

Here are a few examples of images that I took with DSLR. I have used both Canon 6D and Sony A9 over the years.

Image
Micrasterias by Håkan Kvarnström, on Flickr

Image
Micasterias by Håkan Kvarnström, on Flickr

Image
Copepod by Håkan Kvarnström, on Flickr

Image
Cyanobacteria - Gloeotrichia by Håkan Kvarnström, on Flickr

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75RR
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Re: For fluorescence microscopy, specialized microscope camera or DSLR?

#7 Post by 75RR » Mon Aug 10, 2020 4:48 am

Re DSLR: As they say "The proof of the pudding is in the tasting." Those are really excellent images

The last one particularly is spectacular!
Zeiss Standard WL (somewhat fashion challenged) & Wild M8
Olympus E-P2 (Micro Four Thirds Camera)

MichaelG.
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Re: For fluorescence microscopy, specialized microscope camera or DSLR?

#8 Post by MichaelG. » Mon Aug 10, 2020 8:04 am

75RR wrote:
Mon Aug 10, 2020 4:48 am
Re DSLR: As they say "The proof of the pudding is in the tasting." Those are really excellent images

The last one particularly is spectacular!

I agree completely

MichaelG.

.

P.S. For clarity: My reason for linking Louis Keal’s presentation is that whilst most of us are primarily interested in the beauty of fluorescence, there are others for whom it is an analytical tool ... and it is difficult [perhaps impossible] to do quantitative work on images from DSLR, or other general-purpose cameras.
Too many 'projects'

Leitzcycler
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Re: For fluorescence microscopy, specialized microscope camera or DSLR?

#9 Post by Leitzcycler » Tue Aug 11, 2020 1:44 pm

The Rising cam offers 20 MP and is the best dedicated camera I have tested so far - its available on Ebay for about $800.
Could you further specify the model of the best camera as there seem to bee several different models of Rising cam? I have also the problem of selecting a fluorescence/multi purpose camera. However, the DSLR suggested here seem to be very expensive to me.

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