Inverted fluorescence microscope for cell culture

We are currently in need of assistance in identifying whether there exists a variant model of this inverted microscope that is compatible with cell culture plates and flasks.
The distance between condenser mount and slide-stage is very small to insert a cell culture plate or flask.
We would use a 20X objective.
We would greatly appreciate any guidance or recommendations you could provide us with.

Hi @Pritesh, for a fluorescence microscope you would usually use a reflection-style arrangement, so the illumination is from below as well as the imaging. The parts for that are in the complete set of STLs in the customisations and alternatives sections of the build instructions, although the assembly is not well documented there. Instructions for assembling the reflection/fluorescence optics module are most complete for the Delta stage OpenFlexure Delta Stage.

The parts for the optics module for the Delta stage and the Microscope are rotated slightly differently, so you need the appropriate STLs, but the assembly and operation are the same.

For 20x magnification you should be able to focus through about 1mm thick glass base of a dish or flask. There are specific objectives available up to 40x that are designed to account for that 1mm glass thickness.

Several people on the Forum have posted about bright field imaging of cell cultures, their threads may also help.

I believe you should be able to fit a culture plate with no lid. However, you may need to utilize a different objective. A flask presents more of a challenge. Just thinking out loud here… You may need to build a taller condenser tower and perhaps modify the light source to be stronger, or you may use a shorter flask altogether. A chambered slide should work as well.
I have no experience working or building a fluorescence microscope with the OFM platform. But it should be possible.

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For a 20x objective, the condenser lens is useful but not that critical. The ideal thing to do would be to decrease the distance between the condenser lens and the LED, which would then increase the working distance of the condenser. That should be possible by modifying a few parameters in the code, but I’m afraid I might not have time to do that immediately.

However, as William says for fluorescence you’d probably want to epi-illuminate with a beam splitter, so you don’t need a condenser at all for fluorescence imaging.

If a bright-field condenser is useful for setting up etc, a quick and dirty work-around might be just to remove (with a hack saw) the little tube on the condenser that holds the lens - you’d end up just with a bare LED illuminating the sample from above. This won’t be as bright, and image quality will drop slightly, but for a 20x objective it should still work well enough to find your cells before switching to fluorescence.

If you remove the condenser lens, this will give you greater working distance - and as the light wouldn’t be focused any more, you can just raise the condenser a bit if you need to.