Using plastic for microscopy? (View forum version)



jaknight

Posted 11 November 2009 - 01:51 PM

I am working with a cell type that differentiates very well on plastic plates that are made of polystyrene, but differentiate very poorly when grown on glass coverslips. I need to do confocal microscopy with these cells and I am not sure what to do.

I have found plastic coverslips that have the same refractive index of glass and are the same thickness, but I have heard that plastic can autofluoresce and create quite poor images. The other problem is that although these coverslips are plastic they are not polystyrene, so they still might not work for my cell type.

I have tried chamber slides (which also are a different kind of plastic) and my cells differentiate relatively well but still not great.

Another alternative is to try coating glass cover slips with something and see how that goes.

I was wondering if anyone out there has had experience with this sort of thing or can offer advice on how well plastics work for microscopy or has another suggestion.

Thanks.

skeuos

Posted 11 November 2009 - 02:33 PM

I am working with a cell type that differentiates very well on plastic plates that are made of polystyrene, but differentiate very poorly when grown on glass coverslips. I need to do confocal microscopy with these cells and I am not sure what to do.

I have found plastic coverslips that have the same refractive index of glass and are the same thickness, but I have heard that plastic can autofluoresce and create quite poor images. The other problem is that although these coverslips are plastic they are not polystyrene, so they still might not work for my cell type.

I have tried chamber slides (which also are a different kind of plastic) and my cells differentiate relatively well but still not great.

Another alternative is to try coating glass cover slips with something and see how that goes.

I was wondering if anyone out there has had experience with this sort of thing or can offer advice on how well plastics work for microscopy or has another suggestion.

Thanks.


I used plastic coverslips once and it was a complete wreck. Autofluorescence, difficulty focusing, etc. Thermanox #174950 in case you want to avoid them.

I'd look at turning to a substrate. Is there a known substrate your cells will differentiate on?

jaknight

Posted 12 November 2009 - 03:57 AM

I'd look at turning to a substrate. Is there a known substrate your cells will differentiate on?


I am using C2C12s which are a secondary myoblast cell line. Primary myoblasts require collagen coating so I may try that for my cells.

I'll avoid the thermanox, thanks. The chamber slides I am trying are permanox, from the same company. I'll find out today how well they work...

skeuos

Posted 12 November 2009 - 08:12 AM

I am using C2C12s which are a secondary myoblast cell line. Primary myoblasts require collagen coating so I may try that for my cells.

I'll avoid the thermanox, thanks. The chamber slides I am trying are permanox, from the same company. I'll find out today how well they work...


I've used primary myoblasts for similar application, and collagen coating chamber slides worked well enough for IF. Some slides washed off, but perhaps a longer collagen coating period could fix that.

fysio lab

Posted 18 January 2010 - 05:40 AM

hey i'm working with glass CS coated with gelatin 2% solution. Just incubate 30', let the CS dry and then put your dilution of cells on the CS. Worked well for confocal..no background
Good luck