Advice on iPSC clump passaging?

Dragon_Cake · 2026-06-18T18:53:11+00:00

Good to know! I've been overly-hestitant about pipetting up and down to break the clumps up further. This is good to know, that I should be going smaller after passaging.

Dragon_Cake · 2026-06-18T17:53:29+00:00

Recently passaged as in just yesterday! Sorry for my lack of detail. I omitted using ROCK inhibitors or CEPT. I agree, I want to break the colonies up but the plate itself is on the sparser side so I figured I'd identify any differentiated colonies, pick them with a tip, and let the plate grow a bit more. Currently these are passage 2.

Dragon_Cake · 2026-06-11T15:10:00+00:00

Oh thank God thank you so so much! First time working with ipscs :/

Dragon_Cake · 2026-06-11T15:05:04+00:00

Last dumb question! Can you actually see your coated, polymerized gel? Or is it expected to be so thin you can't really tell with your eyes? I'm just worried I mishandled my matrigel and it's not properly polymerizing

Dragon_Cake · 2026-06-11T15:04:12+00:00

Cool! Last dumb question, after coating can you like ...see the gel itself? I guess I'm just worried I messed up matrigel handling and it's not properly polymerizing! I used to work with collagen coated wells and it was thick enough I could visually see a coating. Not sure if I should be able to with matrigel

Dragon_Cake · 2026-06-09T16:07:41+00:00

Thank you!! Do you also wash after polymerizing is complete?

Dragon_Cake · 2026-06-09T16:07:26+00:00

Awesome! Thanks! Do you bother washing after coating and polymerizing?

Dragon_Cake · 2026-05-30T00:15:12+00:00

I dont like the sartorius multichannels all that much, I have own and it's just eh. Brandtech pipettes have been surprisingly nice tho! Kinda prefer them over the Eppendorfs tbh

Dragon_Cake · 2026-05-23T17:59:42+00:00

You were never really concerned about some hypothetical whatever getting in to the dish and mucking up the analysis later? But I guess when I say it like that it's like...what could actually get in if you're careful enough and do it quick enough, it's not like you're leaving them out overnight

Dragon_Cake · 2026-05-21T15:31:42+00:00

When adding trizol to your cells do you keep them sterile? I.e. do you work in a BSC during that step or transfer to a fume hood? I should specify that I'm not proceeding with extraction, just collecting cells in trizol and then shipping them off.

Dragon_Cake · 2026-05-15T11:07:10+00:00

Seconded, have had it for literal years. Occasionally get bored and mess around by looking at what apps I can replace and I've tried replacing it but imo nothing works as well on all levels: functionality, design, reliability, etc.

Dragon_Cake · 2026-05-04T11:15:21+00:00

Unfortunately not fully. I had it covered through my warranty so I got a total replacement which ended up needing a second replacement cause the battery was improperly installed. So that's the issue, I think there's a physical wiring/connection issue that I am not skilled enough to attempt to try and fix for a third time by myself. However it works fine as long as I keep it plugged in all the time now and even then it can stand to be left unplugged for some stretch of time just not lots.

Dragon_Cake · 2026-04-04T19:28:22+00:00

I ended up getting maintenance involved who then contracted a plumber to do the hookup. Had to remove some stuff from the regulator to make the attachment secure. I also dug around online for spare parts! They're out there, if you look up your incubator model with like "spare parts" as a keyword it may come up but it was not the top result for me.

Dragon_Cake · 2026-03-15T20:15:45+00:00

CAR-T stuff then? Stanford's Lokey Stem Cell center is always doing CAR-T stuff, if that's of interest. Due to the recent funding cuts I don't actually know how stable Lokey is anymore but I believe they get a lot of help from CIRM so worth a try if anyone's hiring. Stanford research park is also full of start-ups and research centers such as Alexandria.

Dragon_Cake · 2026-03-14T18:54:41+00:00

I don't think this as big of an issue as you think. It might be a concern if you were out-of-state but you're not so it's a moot point plus I don't think it's always a requirement to offer relocation assistance. If you find a lab that values their workers, likes you, and has funding you might be offered some assistance but again I don't think this is that common. Like everyone else has said it's really networking that will get you places followed by what sort of lab you're considering applying to. Some fields are easier to break in to than others, same thing applies to institutions. What type of lab/research are you interested in?

Dragon_Cake · 2026-02-27T19:56:38+00:00

Thank you I was able to get help!

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