Willing to purchase a working design of a fluorescent microscope with 20X and 40X magnification, motorized scan (0.5mm steps), LED/Laser illumination, interchangeable filters, autofocus, 20Mp camera.
Alternatively, I would engage someone who can design & build such a microscope.
There are aspects of this that OpenFlexure designs could do, but I feel that the current OpenFlexure projects probably don’t meet what you need.
From the sounds of it you probably want something much larger than any OpenFlexure system. Something with a larger range of motion, and mechanisms for interchanging objectives and filters etc. As you scale things up in size they become considerably less stiff, so any significantly larger microscope probably needs to be metal.
It would likely be a considerably expensive and long term research project to develop this from scratch. A possible option is to look at openFrame.
Thank you this is helpful. I will check the openFrame. I will also answer your questions later as I am busy right now.
I want to design a special purpose fluorescent microscope that has 20X magnification. One idea I had is to design a illumination sleeve which would go around the 20X objective and contain one or more LEDs or laser diodes pointing at the sample below the objective. Then I would apply a single emission filter at the output of the objective and focus the light onto a digital camera. The challenge is to scan the sample which is about 4mm in diameter. I would also want autofocus. Changing things on the fly is not needed. We would select the objective and light sources during experiments but once we figure the right combination of objective +light source + filter we would stay with the one that has best results. openFrame is an overkill I think.
I am looking for someone who could help with the design, especially the autofocus, taking automatic photos and uploading them to a cloud.
Perhaps you know of open source components I could use.
I use fluorescent microscopes for diagnosis and research and this is a really interesting idea. I guess that in theory it should be possible but it will require a lot of work, validation, redesign, and investment. Search no more!. Julian @j.stirling is the only man in this planet that can make something like this work. Then you have to consider the cost of the components. Fluorescent filters ain’t cheap. My “guesstimation” is that it will cost between $2000-$3000 USD in components alone and you will put yourself in a rabbit hole spending more money to make it work. At one point I researched theater light gel filters and cheap LEDs but I am not sure whether this approach will work at all.
If the total sample is 4mm across then OpenFlexure has 12mmx12mm range, so it should be fine. Though it will take some time to capture the whole sample.
@JohemianKnapsody and @dgrosen can tell you all about the scan and stitch functionality of the OpenFlexure. Getting it to automatically upload to the cloud should be possible.
OpenFlexure has autofocus, the autofocus algorithm may struggle with fluorescence images due to the dark background, this is because the current autofocus algorithm uses JPEG compression size as a metric for focus (as the compression is done on the GPU making it fast). A paper is available on the algorithm that we use.
Fluorescence has been done with OpenFlexure both for the OpenFlexure Microscope and for the OpenFlexure Delta Stage. The hardware for epi-fluoresence and for the delta stage haven’t gone though anywhere near as much testing as the microscope has, and the code could do with a lot more tidying, to make it easier to develop in the long term.
It may be possible to illuminate from beside the objective. The issue here will be getting uniform illumination. Also I think seeing as the light is coming it at an oblique angle over a very large area this will create more unwanted background. Also by illuminating the whole area you may photobleach areas before you have time to image them.
As for filters, I think whether you can use lighting gels or need more expensive filters will depend on the interaction you are looking at and how far separated the excitation and emission wavelengths are. I know that the Enderscope used lighting gels.
Overall, the OpenFlexure designs should get you one the way to doing what you want. But I think it will certainly be a research project rather than a standard build from the instructions.
I already build the OpenFlexure Microscope and used it but want to rebuild it, so it´s capable to imaging the epi-fluorescence. You said that you already done it and it works with the normal OpenFlexure Microscope. I want to print the needed plastic parts for it and want to ask, if it´s possible to get the STL-files you used and whether you can give some good advice for it? It would help me a lot.
Thank you.
Hi Til,
We don’t have complete instructions for rebuilding the microscope in Epi-fluorescence mode, but what we have is:
I think this should be everything, but @JohemianKnapsody may correct me.
It is also worth noting that while we have done fluorescence on the microscope in the past, it is not as tried, tested, robust and documented as the standard transmission microscopy modes.
One other thing, you will need to source filters. The delta stage instructions have a little on this, but the ones you are buying will depend on your application, and your location. There was quite a long issue thread on this in the microscope repository in the past.
Happy building! Let us know how it goes.