I've been culturing mixed glia and getting microglia from them for years, but it's always struck me that they look slightly yellow under the microscope. If you take a lysate of microglial cell as well, if you look at it, it is yellow-tinged too! Does anybody know what it is that the microglia are full of that gives them this colour? I've never seen this with any other cell type. Thanks!
I ve cultured HAPI (microlgial cell line) a couple of times before..and i found the media turning into yellow color on the very next day..or after few hours.. since its a type of aggressively growing cell line, changes ph of media very easy. But the cells were ok..didnt change the color in my case. In your case may be its due to delay of changing the media before it turned yellow as its possible to get them into the cells I guess.
Hi Yuan! The most common method is to prepare a mixed glial culture using postnatal rodent brain, leave it to establish for 7-10 days and then shake off the microglia. This is the classic protocol of McCarthy and de Vellis, 1980 and reproducibly works very well. To do this you'll need a shaking incubator that you can shake the flasks at about 100rpm overnight with. Then just recover the media, count the microglia and plate out. You could also do more complicated techniques like, isolating the microglia with antibody, eg CD11b, coated beads, but the simple method works well! Hope this helps, happy to answer any more questions you might have. Marc
Hi Rubina, thanks for your answer. It's actually the primary microglia themselves that are yellowish, rather than the media. The must contain some pigment, but I don't what it is! If you look at the cells they always have this colour, even when very fresh and healthy. Good luck with your ongoing research Rubina!
No problem at all! Don't worry about my comment about the microglial colour, that is just a side observation I've noticed over the years! Are you using rat or mouse microglia? Both can look different in vitro, especially in the first few days after plating.
Hi Marc. That is interesting! I have never noticed the microglia being yellow. I wonder if it is due to their high expression of the NADPH oxidase, which contains FAD that should be yellow. I think this contributes to the yellowness of neutrophils. It might be worth taking a spectrum, and comparing it to FAD and FMN. These are also fluorescent, so might be worth taking a fluorescensce spectrum. If it does not fit a flavin spectrum, you could compare it to lipofuscin?
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