I am thinking of trying a LMP agarose, after adding a protein solution in PBS, and incubation at 4 degrees overnight. There is also polycaprolactone which seems to require quite some chemicals that we don't currently have. I am also not happy with what comes out with collagen - very uneven coverage on a clean coverslip. So, I am looking for more simple options:)
While I am certain you are not doing any plant cell culture, other options to look at would be phytagel and gelzan, both of which will actually dissolve and gel at room temperature (with lots of aggressive mixing) and become fluid at your specified temperature interval. They also display very good spreading (as that seems to be a concern), while melted. Both are relatively inexpensive for specialty gelling agents.
No, animal cells only. Looks like phytagel and gelzan are derivatives of agar, right?
Nope, gellan gum. It is an agar alternative, not derivative, and contains no agarose, but is a polysaccharide. As far as I know it has been judged suitable for human consumption, and has been shown to be "non-toxic" on the human organism level. Whether it is suitable for your cell culture is yet to be determined. LMP agarose might be the cheaper option, but phytagel and other gellan gum products have better translucency for microscopy applications, even at higher percentages (use half the amount of gellan gum as agarose to get the same gel stiffness). One can observe single cells in quite good detail on an inverted microscope through about 15-20mm of the stuff. However, how your protein solution will interact with it is yet to be seen, it is anionic, so basic proteins may aggregate once mixed into the gel, which will potentially cause some observation problems (among the other obvious problems). It would be quite interesting to see someone apply the stuff to animal cell culture.
Ok, I will update if I try agar or other products for my cultures. From a publication, it looks like, with -OH groups on the surface, all you have to do is dissolve a matrix protein in PBS, pour onto gel, and leave overnight in the fridge at 4 degrees.
Methylcellulose?
Methylcelluose solutions when heated will reversibly form gels at a temperature characteristic of the type and percentage. For these products, a 2% solution (w/w) has a gelation temperature of approximately 48 °C.(from Sigma's METHYL CELLULOSE
Prod. Nos.M0262, M0387, M0512, M6385 and M7140). It's kind of tricky to prepare.
Hope this helps.
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