Hey,
DAPI/Hoechst excitation require UV light, so if there's a UV filter in front of your camera, you'll see only the fluorescence signal.
However, common glass is not transparent to UV, so it might be that if you're using cheap glassware in the beam path you'll have a problem.
The 100X objective does not let much light go through it, but Hoechst and DAPI have a very high signal-to-noise ratio, they're very bright, so even in an hacked system there shouldn't be much of a problem.
You can find microscope cameras around, but does your microscope allow for a camera? Is it a trinocular? You can spend additional money for a camera and a C-Mount and hack light source and filters, but probably at that point the cost becomes too high.
If you wanna go DIY, then I would hack the system completely. You can just stick a raspberry Pi camera in front of the ocular (possibly with a 20X magnification if you have it) and it should come with a UV filter (I don't know how much useful).
There are UV leds that you could use for illumination, you have to make sure that they would provide enough light.
Filters in your case is easy, you can find UV filters.
The solution with the UV light source in one ocular makes sense and it's probably the only one available to you (beside the paralens): you need the light to point away from your eyes and have the fluorescence reflected back, differently to what you are probably used to in brightfield.
Best,
ukitel
On Saturday, 24 June 2017 09:36:52 UTC+2, Mike wrote:
Abizar,
Thank you - yes I'd be very interested in trying the setup you suggest - please can you post an image? I have Zeiss Standard 25 binocular microscope. The key thing is that I need a sharp fluorescence image with a 100x oil objective.
- Is the setup you suggest compatible with this type of objective?
- Where can I get the required parts and how do I make them fit my microscope (LED, filters, camera)?
- Can I acquire LED lighting that will provide for UV illumination (for use with DAPI, Hoechst)?
Roninlaw,
The price of the core Paralens Advance system with a single objective lens type is approximately £900 including VAT.
Thanks guys,
Mike
On Friday, June 23, 2017 at 11:15:21 PM UTC+1, Roninlaw wrote:
Price?
On Fri, Jun 23, 2017 at 2:25 PM, Mike
<alanmb...@gmail.com> wrote:
Hello all,
As a cheap alternative to a fluorescence microscope for my home lab, I am considering the Paralens Advance screw-on objective lens.
(Please see page 9 of the PDF for an illustration of light spectra)
The Paralens produces excitation at 385-480nm and emission at 480nm (as far as I can tell, please see PDF above!). [b]Question[/b]: Will this device work with Hoechst 33258 and DAPI stain (spectra as below)?
- Hoechst 33258: excitation: 352, emission: 461
- DAPI: excitation: 358, emission: 461
Thank you,
Mike
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