I would like to know if anyone has tried microfluidic chip experiments - particularly fluorescence imaging on an upright microscope. What are the parameters that we need to keep in mind to choose an objective as I need to work with high magnification (60X).Or if anyone has tried placing and imaging a microfluidic chip in an upside down manner on an upright microscope using some sort of adaptor so to still use a high numerical aperture objective?
Thanks
High NA usually means short working distance. This is often problematic for fluidic devices that are intubated on one side and mounted on something rigid, like a glass slide or coverslip across the entirety of the other side.
We have used custom fabricated acrylic holders for this purpose in the past. They allow you to have a path for the tubing to exit out one or both sides while allowing your objective to come as close as necessary to the glass which is now on the top side. The setup is essentially like a really thick slide (10-12mm). Keep in mind that some objectives have working distances of less than a typical slide (1mm) in which case you'll have to mount your fluidic on a cover glass instead.
the same as Ron here, You can use an upright but you will need to mount the chip upside down. depending on the holder plate you can even pass the tubing trough the holder plate opening, and then use standard PDMS punchs.
If you use short working distance objectives, use #1 thickness coverslip, and be carefull not to beak it. An machined or 3D printed adapter could help.
Hi Thanks Ron and Benoit for the answers.
@Ron and Benoit - So if I understand correctly you have placed the microfluidic chip upside down on an adaptor (custom made?) Co you let me know from where I could get these custom made adaptors from?
Thanks a lot.
I just posted a simple holder to Thingiverse. https://www.thingiverse.com/thing:3480785
Does your institution have a machine shop? Do you have access to a 3D printer? This design is simple enough to make by hand. If you use acrylic, don't machine the two polished faces of the sheet and this will insure good flatness of the fluidic device on the stage. Any other machinable materials may be used. Polycarbonate and polyethylene are good options too.
No I do not have access to a 3D printer. but a machine shop maybe yes. Thanks so much again. Also, If there are any commercially available adaptors please let me know. Thanks.
https://bmcmolcellbiol.biomedcentral.com/articles/10.1186/s12860-018-0158-z
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