I need to glue a functionalized coverslip with Valap to the bottom of a chamber. Coverslips are 22x22 mm, #72198-10, from EMS.
1. Does anyone know of commercial products available for that size? I mean a sort of insert or chamber, where you can mount these coverslips.
2. I also have an option of machining. At our facility, they agreed to make the chambers from aluminum or 3d print with ClearVue. My cells go into apoptosis about 1 day after seeding in a Clearvue chamber. They do attach and spread before that on the coverslip. What does everybody think of these materials?
I can also find a way to do that with PDMS. Does anyone have experience growing cells in big PDMS chambers?
Thanks.
I assume there is a reason why you can't simply put the coverslip in the bottom of a standard tissue culture dish and seed the cells into the dish/on top of the coverslip. What do you plan to do with the coverslip once you plate your cells onto them? Are you looking to perform live-cell microscopy through the coverslip? If so, if you have access to a machine shop, can't they just drill a suitably sized hole into the bottom of standard 35- or 60-mm tissue culture dishes for you to glue your coverslip over? If they can't do that, I think Mat-Tek can provide pre-drilled dishes.
I assume there is a reason why you can't simply put the coverslip in the bottom of a standard tissue culture dish and seed the cells into the dish/on top of the coverslip. What do you plan to do with the coverslip once you plate your cells onto them? Are you looking to perform live-cell microscopy through the coverslip? If so, if you have access to a machine shop, can't they just drill a suitably sized hole into the bottom of standard 35- or 60-mm tissue culture dishes for you to glue your coverslip over? If they can't do that, I think Mat-Tek can provide pre-drilled dishes.
I use a high NA objective which doesn't allow me to use anything underneath the coverslip and image cells. Thanks for the idea of drilling, I will ask!
Thank you everyone! Drilling worked quite well, only Valap becomes leaky after 2-3 days in a medium. But that is a different issue.
We mainly used Dow Corning high vacuum grease. For no particular reason except that we had it in the lab and used it for lyophilizers and other purposes. But it works very well with cells in culture and doesn't seem to have any toxicity.
The standard procedure for gluing primary culture explants is to place the tissue onto a tiny droplet of serum on the dry plate surface, then let dry a few minutes in a sterile hood before adding medium; whole serum is an excellent glue (and far less risky for impeding growth and activity than grease). This method has also been used to transfer explanted cells growing out on coverslip chips. Gelatin can be used instead of serum, but because of its gelling properties a low-temp agarose is more appropriate.
The tiniest droplet is sufficient to be drawn by capillary action along the interface of your coverslip and the dish/chamber, and you can suction any excess with a tip of sterile filter paper. You can use your standard labware, no drilling or refractive grease-bearing interfaces involved.
- Badges
- Science method