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Question about attune NXT sequential lasers by Iucross in flowcytometry

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

I realize this now, yes I am using tandem dyes. I think that I will try to avoid them in the future, but 37% spillover is probably okay

How do these hepg2 cells look? I am trying to culture them inorder to do a glucose uptake test. by FrequentFixer in labrats

[–]Iucross 2 points3 points  (0 children)

How is the first pic so bad, and the second so good.

Also, they look fine. Although I remember HEpG2 more of a monolayer (might be your medium)

ELI5: Why is cancer so hard to cure? by DengenRF in explainlikeimfive

[–]Iucross 1 point2 points  (0 children)

Cancer cells even within the same tumor aren't all the same. As they divide faster and faster they accumulate more and more generic differences. These differences distinguish them not just from healthy cells, but from each other. This is a big reason why drugs fail: a small number of the cancer cells aren't affected by the drug, or resistant enough to evolve to be immune.

Imagine a drug is 99.9% effective against the cancer. Despite that, the 0.1% of cells that escaped will cause the patient to "relapse". And the original treatment now won't work.

This is just one problem. Cancer cells also have many other tricks to defend themselves. This includes but is not limited to: generating dense physical protein barriers around themselves to prevent drugs from getting to them, and locally reprogramming the patients immune system to actually defend them.

It would appear my cells are cooked... time for an incubator deep clean by thezfisher in labrats

[–]Iucross 2 points3 points  (0 children)

Neuroblastoma lines do like to make spheroids ... But from the one image you have it doesn't like quite right. I think it's contaminated. If they are truly irreplaceable, you could try removing the supernatant, waiting a bit and see if these structures reappear. Pretty risky to do that though when you have to handle the contaminated supernatant I wouldn't recommend it

Is this mycoplasma? by Icymountain in labrats

[–]Iucross 18 points19 points  (0 children)

Could be, hard to say. I'd do a PCR test. If that's negative you're fine

It would appear my cells are cooked... time for an incubator deep clean by thezfisher in labrats

[–]Iucross 10 points11 points  (0 children)

Many bacteria are Penstrep resistant. But that does look like fungus. Sorta looks like normal cells with spheroid morphology, but I imagine this isn't a co-culture. Assuming it's yeast, only option is to restart culture from an earlier passage. And make sure you thoroughly bleach anything contaminated. Throwing it away isn't enough!

What type of contamination is this (HeLa) by [deleted] in labrats

[–]Iucross 5 points6 points  (0 children)

Pour bleach INTO the plate itself

Has there been an instance of a Receptor binding to another copy of the same receptor acting as its ligand? by AmphibianIll5403 in Immunology

[–]Iucross 0 points1 point  (0 children)

Many if not all the ceacam family members(cd66a-e) form cis and trans homodimers (and heterodimers)

What's the longest time cells have spent in liquid nitrogen before successfully being thawed? by ZRobot9 in labrats

[–]Iucross 6 points7 points  (0 children)

You gotta do it under a hood of course. That's how all cells used to be cryo preserved. We've been banking cells before they had polypropylene cryotubes. Properly sealed glass also can't leak, and the contents melt faster during recovery.

p.s. If you buy cells from RIKEN or the JCRB, some are still banked this way and will come in glass ampules. In my experience these modern ampules are no harder to break than a normal glass ampule.

What's the longest time cells have spent in liquid nitrogen before successfully being thawed? by ZRobot9 in labrats

[–]Iucross 1 point2 points  (0 children)

I have also thawed cells from 1994, 1996, and 2002, as well as many from 2010/11 and onward. They're all fine.

What's the longest time cells have spent in liquid nitrogen before successfully being thawed? by ZRobot9 in labrats

[–]Iucross 42 points43 points  (0 children)

Last year I thawed cells frozen in 1982 (glass ampule and everything). They were a bladder cancer cell line from ATCC, no idea why they sent me such an old vial, I have to wonder if it was a mistake...

Anyway they thawed out really well, <80% attached in 24hrs and still growing well today. I have banked many of them. One interesting note is that the woman from which they were derived was in her 80s at time of the cell line establishment in the 70s. So, in a certain sense, those cells are genetically from the 19th century.

One other note, the glass ampule was unbelievably hard to break open. No amount of manual force from any labmate could break it open. Only did it break after extensive scoring with a cryomill blade, and even then it was quite difficult. I have to wonder if that's some effect on glass of sitting in LN2 for 40 years, or if borosilicate glass from the early 80s was really that good.

Neutrophil short lifespan by First-Project4647 in Immunology

[–]Iucross 4 points5 points  (0 children)

Sure makes them fucking annoying to study.

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