I was using phenol red free neurobasal media for live cell imaging on cultured primary neurons previously and it works very well (for many hours on the confocal microscope)
Recently I found a new batch of that medium will cause the cell death even in the cell culture incubator before I started the experiment.
I was wondering if anyone could suggest me an alternative media that I could use as I have some very critical experiment to do at the moment.
Thanks a lot.
UPDATE: I ended following Rodolphe Hajj's suggestion using the normal Neurobasal medium with phenol red for imaging a few years ago. Although the background is a bit high but it is acceptable and will disappear quickly by photobleaching effect.
Dear Di Xia,
You haven't mentioned if your microscope is equipped with heating system and CO2 supply to keep cells alive while imaging. If it is not the case then you can add either HEPES to the medium to keep the buffer or use CO2 independent media available with life technologies. Using HEPES will keep your cells alive for only few hours and it depends on the concentration used.
Good luck.
Rajesh
Hello Dr. Xia,
We use a normal Tyrode solution when we are imaging without a CO2 supply. As pointed out above, it is HEPES based and we are able to image primary neurons with or without a heating stage for several hours while maintaining good viability.
Best,
Kristina
Keep in mind that the osmolarity will have a profound effect on the health of your cells and, consequently live imaging. If you were using Neurobasal (for embryonic cells) then the osmolarity is very low (~230 mOsm) and you will have to adjust your recording medium accordingly. Best to check all solutions with an osmometer
Normal Tyrodes. Easy and cheap to prepare. Setting the pH can take some time to set but you can always make a stock solution and save time on the whole. Add glucose only when you make dilutions.
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