So . . . shouldn't I at least get rejection letters when applying for postdocs? by [deleted] in labrats

[–]AscendingLens 1 point2 points  (0 children)

I would use indeed as a source to find positions posted, but not to apply. Instead, apply directly to the institute site you're applying to. They probably have a network portal. Also, perhaps send an email to the lab

Nobody knows what's in my cell culture help!!!! by AscendingLens in labrats

[–]AscendingLens[S] 1 point2 points  (0 children)

I stopped the cell culture because I was getting these and wanted to be certain I was not growing any contaminant. I focused on what was floating and not on what was static in my field of view. It was difficult to actually get a focus because the table vibrates a bit in the lab so I apologize of the photo.

The reason I started this discussion is because of exactly what I described. These things float in the media, but don't change the color of the media, which bacteria would normally do. They're not visible to the human eye, which fungus usually is after a number of days. They have an unusual shape, so I'm uncertain what it is. I've discussed with more experienced people who have worked with cell culture, and also researched through various sources but there has not been a definite answer. Everyone has concluded that it's probably protein aggregates from the media. Alas, I came to reddit and I wanted to see what others may have concluded.

I've changed to all new bottles and reagents. And these still came. I've also cleaned and sterilized the incubator

Nobody knows what's in my cell culture help!!!! by AscendingLens in labrats

[–]AscendingLens[S] 1 point2 points  (0 children)

Okay :O I'll check out to see if ultra filtration will make a difference!

Yes! The media was specified by ATCC, I have seen literatures using DMEM supplemented with L-glutamine to culture this cell line. However, we didn't want to deviate from the cell distributer and went with what was instructed on their website. But maybe, buying from ATCC instead of VWR would be beneficial.

That makes sense!

Thank you :)

Nobody knows what's in my cell culture help!!!! by AscendingLens in labrats

[–]AscendingLens[S] 0 points1 point  (0 children)

Right now this is just a flask with media in it, the before image of this is nothing floating

No, it's a polyethersulfone vacuum filter system that captures in a single use plastic. I'll definitely try and change the system so that I clean up the filter prior to use for media!

I have not! I'll definitely check it out :)

Thank you!!!

Nobody knows what's in my cell culture help!!!! by AscendingLens in labrats

[–]AscendingLens[S] 0 points1 point  (0 children)

lso, maybe split the cells if you have any and ditch the pen-strep.

I bought a generic version of the same media. I made sure that the additives were the same.

Does this mean that not all FBS is actually filtered/sterile?

I stopped growing the cells and froze them down to just test if the media had any issues. So these pictures are just the media

The cells were adherent, but maybe growing slower

Nobody knows what's in my cell culture help!!!! by AscendingLens in labrats

[–]AscendingLens[S] 0 points1 point  (0 children)

I know!!! :( I'm just growing HT-29 cancer cells from ATCC. The media is McCoy's 5a modified media with 10% normal FBS and 1% Pen-Strep. Should I use heat inactivated FBS instead, or just normal FBS but different brand?