I'm culturing mamospheres cancer stem cells and they are floating in the media. How can I help their attachment to perform the cytoskeleton staining or if there is another way to do it?
You can also stain them in suspension in buffer or Cygel and then seed on the microscope slide. Be careful with the cover slips not t squeeze them. you can use some drops of nail polish (!!) around your samples to place the coverslips not squeezeing spheres, that it takes some time to ptimize imaging, but results are better than letting spheres "sit" on a matrix. I mean, it also depend from what you are interested to stain for. good luck!
Hi Abdulaziz,
You can do intracellular staining on free floating spheres, but you will only stain the outer layer of cells, In most cases that is fine. We do it all the time. We do the staining as normal then place the spheres in a glass bottom dish for viewing on the confocal. They will settle to the bottom and can be imaged easily no need to use gel!
Kind regards,
Steve
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