DIY Epifluorescence Macroscope

Hey Microscopists,

I recently designed and built an epifluorescence macroscope centered around the Olympus XLFLUOR 4x/0.28NA objective and Kinetix camera for both fixed and live imaging. It uses the high power CBT-90 LEDs driven at 27 amps as the excitation source to get up to 690 mW (29 mW/mm^2) of excitation light onto the sample plane for very high temporal resolution imaging.

Preliminary tests showed that the system is working very well for us so I figured I would share the CAD files with anyone who may need a similar setup: https://github.com/Llamero/DIY_Epifluorescence_Macroscope

Specifically, this macroscope has a 4.7 mm x 4.7 mm FOV and the optical resolution is pixel limited with the 10 MP Kinetix camera; having pixels that are equal to 1.5 µm x 1.5 µm at the sample plane. The end result is a nearly 5 mm FOV with subcellular resolution!

If high temporal resolution is not needed, the custom LED source can be replaced with any standard microscope illumination source that uses a 3 mm liquid light guide, and the Kinetix camera could be replaced with an astronomy camera with a sensor that is at least 22 mm x 22 mm, such as this camera: https://www.bhphotovideo.com/c/product/1417197-REG/starlight_xpress_trius_sx_46_usb_hub.html

If you have any questions, please feel free to ask.

Cheers,
Ben Smith

P.S.
The development of this scope and illuminator was funded by the following grant: NIH P30EY003176

Also, definite shout-out to Thorlabs for selling screwball parts such as the 60 mm filter cube and tube lenses with massive apertures, which make this sort of system even possible.

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Here is the first test image we got off the scope; this is an image of
endogenous tdTomato fluorescence in a PFA fixed Opn4-Cre/+; floxed
tdTomato/+ P4 mouse retina (link to full resolution raw tiff): Opn4-Cre - tdTom P4 retina.tiff - Google Drive

The image was taken in dynamic range mode (16-bit) with a 50 ms exposure.
The retina wasn’t perfectly flat, so some parts are in focus while others
are a bit out of focus, which is a testament to the high axial resolution
of the 4x objective.

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