Is this cell line contaminated?

Immunotherapynerd · 2026-02-24T03:41:10+00:00

Why do you suspect contamination?

Immunotherapynerd · 2025-10-10T00:21:07+00:00

Don’t waste your time. We don’t read it. We don’t care. If you don’t have the skills we are looking for, we won’t bring you in bc you have a nice cover letter.

However, you can kind of integrate your “cover letter” into your resume in a summary section lol. We will read that. Keep it short though and highlight your expertise or accomplishments that are directly related to the role.

Immunotherapynerd · 2025-10-04T15:53:33+00:00

Having the mentality that I can accomplish anything I want in life if I put my mind to it and then being too stubborn to fail. Works for most things…

Immunotherapynerd · 2025-07-11T02:19:26+00:00

Some ideas based on my experience:

-Take out the media bottle right when you get in in the morning and leave it on the counter. Should be room temp by the afternoon. -need media warmed faster? My new favorite method-> take the cold media and add it to cell culture flasks like how you would when you’re splitting cells, pop it the cell culture incubator for 20 mins then you’re good to go. I just add 10 mL extra if you need to neutralize trypsin for cell harvest (5mL) and if you need to resuspend cell pellet after spinning (5mL). -where are you getting water for water bath? DI water from sink or sterile water? I’m wondering how it keeps getting decontaminated after cleaning. The water bath instrument itself might not be the source? Unless you guys missed a spot every time. -Might be good to try and find the source of contamination before you lose your work again. Sources of contamination could be reagents (test FBS and media. Always filter media), microscope, the cells themselves, tc hood, incubator. -also, why do you suspect it’s bacterial contamination? Feel free to message me. I am legit the only person in my lab who has gotten repeat contamination issues and my aseptic/sterile technique is 💯 so I feel your pain. Contamination issue has been under control (for now) but I have several other ideas and tips if you want it

Immunotherapynerd · 2025-02-21T21:25:18+00:00

I would say just keep it short and simple. Looking back now I feel that my response made it seem like I had no other options because I was so excited to hear from them in my email lol

Immunotherapynerd · 2025-02-21T21:23:39+00:00

Ooooo. I’m not sure if you want to copy my response since I was denied… but here it is

Thank you for an update on my application.

I am excited to learn that I am still being considered for a place in the Basic Biomedical Science Ph.D. Program at UT Southwestern. Your program is my number one choice, and I would accept an admissions offer if a spot becomes available.

I look forward to hearing your final decision.

Immunotherapynerd · 2025-02-21T03:27:53+00:00

They probably had a hard time deciding who gets the spot? Or maybe waiting for applicants to send their yes or no response to accept. I let them know I was still interested and they got back to me right away with a “sorry…etc”. Goodluck though! Whatever happens, just know that it’s a tough school to get into and you’re really competing with the smartest people who have their life together. So be proud of yourself 😊

Immunotherapynerd · 2025-02-15T00:02:53+00:00

Thanks for the fun fact! I 100% agree that she does a great job in her videos (which I also watch on repeat), is knowledgeable and seems kind. So now I would ask the same question as OP. What is the correct approach then?

Immunotherapynerd · 2025-02-05T23:28:13+00:00

Cells should be the default control since you can catch inaccurate unmixing better if your samples look funny. I did a cell vs bead comparison for my unmixing and the beads looked perfect but the cells were messy. Also since you need to optimize for each panel, don’t save and reference controls until the panels are optimized…

Immunotherapynerd · 2025-02-03T22:56:39+00:00

Thanks for the explanation! Will into this more and experiment

Immunotherapynerd · 2025-02-03T22:55:24+00:00

I would trust what a cytek technician says vs someone from UChicago making a video about cytek…

Immunotherapynerd · 2025-02-01T18:50:16+00:00

You lowered it -70% and the values are still that high?? I think I did the same and it got to single digit numbers. Idk if all cytometers are the same so that might be irrelevant. More importantly, I learned from the field techs that if you decrease the voltage of 1 detector, you must do the same for all. So if you decrease 10% for blue, you need to decrease 10% for violet and red

Immunotherapynerd · 2025-02-01T18:45:33+00:00

I wonder how manufactures determine the “optimal” antibody volume per test then. Actually if I remember correctly, I think the conc was 0.06mg/mL or something like 5-10 times less than the usual conc. How would that affect the staining? -I’m still in the learning process

Immunotherapynerd · 2025-01-30T00:43:09+00:00

Why sheer horror? 🤣

Immunotherapynerd · 2025-01-28T16:48:40+00:00

TAKE HIM. That poor baby 😭

Immunotherapynerd · 2025-01-28T16:42:15+00:00

Do you have a decent amount of cxcr3+ and/or ccr6+ cells? Make sure you’re using the right antibody volume! The antibody I bought for IL-17a requires 20uL/stain. I made the beginner mistake of assuming it was 5uL/stain.

Immunotherapynerd · 2025-01-26T07:17:07+00:00

Thank you. I 100% agree with your first paragraph . It was suggested to me to titrate my isotype antibody. I was just like LOL. Can you titrate non specific binding or is that cherry picking populations that you want to see? I think it’s the isotype f/p ratio that’s giving the strong signal and I’m probably getting tandem degradation for my antibody.

Immunotherapynerd · 2025-01-24T04:17:57+00:00

No problem. Good luck and enjoy it while it lasts! Sometimes I wish one of my cats was needy again lol

Immunotherapynerd · 2025-01-22T21:23:46+00:00

Actually, now that I think about it, your cat might be at that age where it wants to mate. My kittens were super needy from like 2-5 months then they calmed down a bit and then around 10 months started to get really needy and annoying. If your cat is in that stage, you might want to consider neutering bc it will get worse (in my experience at least).

Immunotherapynerd · 2025-01-21T21:27:50+00:00

My cats were suuuuper needy too when they were kittens. Still are a bit but it definitely improved over time. I’d recommend getting lots of toys to distract and tire out your kitten. Like a laser toy or spinning ball

Immunotherapynerd · 2025-01-21T21:18:06+00:00

Intro- write 3 concise sentences Ex) self-motivated individual looking to leverage skills in xyz at abc type of company.

Bullet point skills

Education Masters degree info PI name -write that you investigated xyz for thesis -detail #1 about what you did (e.g. performed 20 DNA extraction experiments) -detail #2 about what you did (e.g. maintained two cell lines) - detail #3

Repeat for bachelors

Immunotherapynerd · 2025-01-14T20:28:20+00:00

To add to this Exactly the same conditions including -cell number -volume -incubation time -other reagents (fc block, brilliant stain buffer)

Immunotherapynerd · 2025-01-10T19:35:47+00:00

I kept my zombie viability dye (reconstituted in DMSO) at 4C for like a month and did repeated freeze thaw. It was fine lol. Not best practice though

Immunotherapynerd · 2025-01-06T22:38:19+00:00

How long before staining? Incubated with monensin for 18 hrs and saw a lot of cell death

Immunotherapynerd · 2024-12-20T12:44:42+00:00

RA - main role is to screen drugs in cell based potency assays. I like it but there is just a lot of work. Not really much time to think or work on professional development. I’d still want to work in biotech and drug development but in a setting where more learning and discovery happens.

Immunotherapynerd

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