Has anyone ever mounted naive THP-1 cells on coverslips for confocal imagining? I want to image THP-1 cells without polarizing them to M0 macrophages, but I have had problems getting them to adhere to the coverslips.
poly-L-lysine treated coverslips.
Good luck with your experiments.
http://www.bdbiosciences.com/resources/protocols/immuno_microscopy.jsp
Did you tried increasing the Poly-L-Lysine concentration? Usually the working standard is 0.01%, but you could increase it for optimal conditions.
Well I tried the poly - l- lysine and the cells were still there after allowing them to adhere for an hour and then removing the media. However i must have lost them again somewhere in between my treatment, washing, and staining
You can try using chamber Chamber-Slide or Chamber-coverglass systems (Thermo scientific or Ibidi wotk fine). Ibidi also has plastic chamber-slides, that should increase the attachment. Works perfect with inverted microscope. I use them for my confocal images. Just stain the cells right in the wells, without detaching the chamber.
I use fibronectin coated cover slips for THP-1 cells. sigma-aldrich sells solutions and has a recipe.
Hello everyone discussing this topic,
Shi-fix coverslips are an easy option for this purpose. Suspension cells attach quickly on these coverslips ( 30 mins) or can be directly grown on them. The slides work very well for Immunofluorescence.
http://www.shikharbiotech.com/products/shi-fix-coverslips/
Hope that helps you if the issue is still relevant to you.
Regards,
Ravindra
How can I attach suspension cells to glass slides for.... Available from: https://www.researchgate.net/post/How_can_I_attach_suspension_cells_to_glass_slides_for_fluorescence_microscopy_investigation#599d5d6eeeae392a6221be9e [accessed Aug 23, 2017].
Suspend in 1% or lower low melting low gelling agarose in ultrapure water and place the suspension between coverglass.
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