Hello Lan Mei ,

You can use PBS 7.4 with 5 mM MgCl2. This works fine for most of the cell lines (robust ones). The cells were happy for almost 90 mins for my case. However, if the cells are still stressed then I would suggest to use warm media only (DMEM/RPMI), not the complete growth media as you might get some fibers from FBS which would require washing before imaging.

Thanks.

Most cell lines will be absolutely fine with lower temperature for many hours. If the cells are in flasks and the laboratories far apart then filling the flask with medium is a good idea for transportation. Small volumes of medium will splash round and the liquid movement may damage the cells. Transporting the cells in an inflated polystyrene carrier boz with a bottle of water at 37c to maintain the temperature will avoid any adverse temperature effects

Hello there. For studying cargo uptake, we try to keep any stress in the cells as little as possible. If the transport time is less than 30min, warm media buffered in HEPES alongwith high FBS can make the cells cosy for sometime. However, if tou need to travel longer, I would recommend using cold media with 20% FBS and HEPES buferring and transport over ice. Upon arrival, gently allow the cells to warm up at RT. Wipe off excess humidity and the allow to stabilize n the cage incubator with CO2. This should enable more reliable cargo uptake data. And, yes please use media without phenol red. It is also recommend to change the media on arrival to the one most appropriate to study the respective cargo uptake. We avoid serum while monitoring uptake, so when transporting we use high serum media and perform the serum starvation, cargo pulse and chase on the destination itself. Hope this helps

Hello

In general, a few minutes will not cause cellular damage. I agree with what was suggested in the previous comments. I would add the use of cell death markers to make sure

Following.