Tissue culturing and feeling sad

[Zumab]

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Hi all,
For those who culture cells for research, if you have ever had contamination in your cells, how do you cope with it? I had some pretty bad cross contamination recently and a month's work of cell work has been lost and I will now have to start over. My mentor already knows about it (she is pretty laid back-and said its okay). Research means a lot to me (something I look forward to-although feeding cells daily can be tiresome). I just feel empty and discouraged. I've had contamination before at a different lab, seen others get it as well but for some reason, I'm just very disappointed in myself and having a hard time finding the energy to pick up from here. :'(

 

[mcloaf]

They're just cells...take it easy.

 

[Pacna]

This sounds like my girlfriend's reaction to everything.

 

[TheWeeIceMan]

Whenever I have trouble coping with the loss of cells, I go to cell therapy.

 

[kevinus]

No backup cells in another incubator? No frozen stock? :'(

 

[Stoptheclot]

One way of coping with the loss of a culture is to travel the world, ask deep and meaningful lessons, learn different languages, become more diverse.

That way, you can be multi-cultured.

 

[mehc012]

Hi all,
For those who culture cells for research, if you have ever had contamination in your cells, how do you cope with it? I had some pretty bad cross contamination recently and a month's work of cell work has been lost and I will now have to start over. My mentor already knows about it (she is pretty laid back-and said its okay). Research means a lot to me (something I look forward to-although feeding cells daily can be tiresome). I just feel empty and discouraged. I've had contamination before at a different lab, seen others get it as well but for some reason, I'm just very disappointed in myself and having a hard time finding the energy to pick up from here. :'(

And just picture...if you were in a mouse model you'd be talking about how '3yrs worth of work has been lost'. At least it's just cells! Easy come, easy go!

 

[meretalking]

Hi all,
For those who culture cells for research, if you have ever had contamination in your cells, how do you cope with it? I had some pretty bad cross contamination recently and a month's work of cell work has been lost and I will now have to start over. My mentor already knows about it (she is pretty laid back-and said its okay). Research means a lot to me (something I look forward to-although feeding cells daily can be tiresome). I just feel empty and discouraged. I've had contamination before at a different lab, seen others get it as well but for some reason, I'm just very disappointed in myself and having a hard time finding the energy to pick up from here. :'(

It happens. Be more careful, spray everything with ethanol, start over. Your mentor is relaxed about it because losing a month of work is par for the course.

 

[lalalapartay012]

Hi all,
For those who culture cells for research, if you have ever had contamination in your cells, how do you cope with it? I had some pretty bad cross contamination recently and a month's work of cell work has been lost and I will now have to start over. My mentor already knows about it (she is pretty laid back-and said its okay). Research means a lot to me (something I look forward to-although feeding cells daily can be tiresome). I just feel empty and discouraged. I've had contamination before at a different lab, seen others get it as well but for some reason, I'm just very disappointed in myself and having a hard time finding the energy to pick up from here. :'(

Strive for improvement! It's normal to be sad/disappointed for a day or so, but try to figure out what went wrong, what you can improve on, and go from there 🙂 Sometimes it might not even be your fault... for example, if someone else's cells had mycoplasma and were in the same incubator as yours... good luck trying to avoid contamination! And it's cool that your mentor is so chill about it. It will all be okay! 😀

 

[crimsonkid85]

Hi all,
For those who culture cells for research, if you have ever had contamination in your cells, how do you cope with it? I had some pretty bad cross contamination recently and a month's work of cell work has been lost and I will now have to start over. My mentor already knows about it (she is pretty laid back-and said its okay). Research means a lot to me (something I look forward to-although feeding cells daily can be tiresome). I just feel empty and discouraged. I've had contamination before at a different lab, seen others get it as well but for some reason, I'm just very disappointed in myself and having a hard time finding the energy to pick up from here. :'(

Are you hairy? Do you have hairy arms? NEVER bring your arms or hands over a plate open to hair....I mean air.

 

[BigBoy99]

Listen buddy, the allied forces bombed the CRAP outta Germany not for you to be wasting cells. YOU COULD OF USED THEM U GOTTA BE KIDDING ME, THE VETERANS WHO SAVED THE ALLIED FORCES FOUGHT SO THAT EVERY MAN COULD HAVE AN EDUCATION AND HERE WE HAVE SOME UNGRATEFUL KID MESSING UP CELL CULTURES. THINK ABOUT HOW THE CELLS FEEL, THEY DIDNT DO ANYTHING WRONG BUT NOW THEY DIE. GOD DAMN IT YOU JUST killed 100M+ CELLS AND U WANNA BE A DOCTOR. DAMN MAN DAMN IT.

 

[GoPelicans]

Wash your hands after you poop you neanderthal.

 

[487806]

Listen buddy, the allied forces bombed the CRAP outta Germany not for you to be wasting cells. YOU COULD OF USED THEM U GOTTA BE KIDDING ME, THE VETERANS WHO SAVED THE ALLIED FORCES FOUGHT SO THAT EVERY MAN COULD HAVE AN EDUCATION AND HERE WE HAVE SOME UNGRATEFUL KID MESSING UP CELL CULTURES. THINK ABOUT HOW THE CELLS FEEL, THEY DIDNT DO ANYTHING WRONG BUT NOW THEY DIE. GOD DAMN IT YOU JUST killed 100M+ CELLS AND U WANNA BE A DOCTOR. DAMN MAN DAMN IT.

 

[solitarius]

This is why medical school admissions is so competitive.

 

[BigBoy99]

Lol, I'm just kidding its not the end of the world, if your supervisor isn't mad it's fine just dont do the same thing again and take proper precautions.

 

[Reckoner]

Listen buddy, the allied forces bombed the CRAP outta Germany not for you to be wasting cells. YOU COULD OF USED THEM U GOTTA BE KIDDING ME, THE VETERANS WHO SAVED THE ALLIED FORCES FOUGHT SO THAT EVERY MAN COULD HAVE AN EDUCATION AND HERE WE HAVE SOME UNGRATEFUL KID MESSING UP CELL CULTURES. THINK ABOUT HOW THE CELLS FEEL, THEY DIDNT DO ANYTHING WRONG BUT NOW THEY DIE. GOD DAMN IT YOU JUST killed 100M+ CELLS AND U WANNA BE A DOCTOR. DAMN MAN DAMN IT.

I don't see any connection to Vietnam, Walter

 

[boomshaketheroom]

Umm you start over..? Thats why you have frozen stocks.

 

[choyphin]

I get the same feeling when I take a LCMS of my reaction, all the starting material is gone, and there is no product.

 

[engineer16]

A girl in my lab had her astrocytes contaminated and she cried right then and there. We all thought she was pretty weird for doing that. How old is your incubator? I don't do cell culture, it is hocum and magic of the highest order, but my labmates who do it are often frustrated with an old incubator, cross contamination from a careless person in the adjacent lab that shares space, etc.

 

[crimsonkid85]

A girl in my lab had her astrocytes contaminated and she cried right then and there. We all thought she was pretty weird for doing that. How old is your incubator? I don't do cell culture, it is hocum and magic of the highest order, but my labmates who do it are often frustrated with an old incubator, cross contamination from a careless person in the adjacent lab that shares space, etc.

well if these were her primary astrocyte cultures that she had to do timed matings for, wait 21 days for the pups to be born, then age the pups to day 3-7 and isolate their astrocytes from their smashed up brains, and since they are primary cells they only divide 3-5 times before senescencing, then yes, yes i can see why she would cry as she just witnessed a month of her work go down the tubes for no good reason.

 

[engineer16]

well if these were her primary astrocyte cultures that she had to do timed matings for, wait 21 days for the pups to be born, then age the pups to day 3-7 and isolate their astrocytes from their smashed up brains, and since they are primary cells they only divide 3-5 times before senescencing, then yes, yes i can see why she would cry as she just witnessed a month of her work go down the tubes for no good reason.

Nah she was an undergrad and she hadn't done the isolation, she was new to the lab. And you're not working for the days in utero, come on son.

 

[winterchill]

There, there OP. I did some tissue culturing a few years back for my thesis in undergrad. Started with HSCs and differentiated them into B cells. Took a couple of weeks. And without fail, I'd take them out of the incubator and notice bacteria in my culture.

First, I'd say you should toss anything you suspect is contaminated and practice some sterile technique. Wash your hands, change gloves or double glove, clean down hoods, use individually wrapped serological pipettes, new reagents, don't reach over open tubes, don't put down caps. Try growing some cells you can afford to lose and make sure those aren't contaminated. You just need to rebuild your faith that your work won't continue to go down the drain.

Second, resilience. Failure is character building and character reinforcing. I hate the phrase "it's not the end of the world" because if it was, there'd be nothing to do about it anyway. But here you have some agency. Work on your technique and understand that these things happen. Your PI isn't upset because it's not that big a deal. It's a bump in the road. You'll hop off your seat and settle back in. Breathe. You can do this.

Godspeed.

 

[organesha]

This happened to me after my first few times in cell culture. I'm just really anal about sterilization with EtOH now and haven't had any issues since I started two years ago. Don't worry too much, it happens to everyone.

 

[baconshrimps]

They are cells.
Research, goes on.
Test for mycoplasm often, if you can.

 

[crimsonkid85]

Nah she was an undergrad and she hadn't done the isolation, she was new to the lab. And you're not working for the days in utero, come on son.

of course you're doing other experiments while waiting. the point is though that in order to repeat her failed experiment she must wait ANOTHER month. and trust me, if your graduation hinged on this experiment you would be crying over the lost month

 

[Zumab]

This happened to me after my first few times in cell culture. I'm just really anal about sterilization with EtOH now and haven't had any issues since I started two years ago. Don't worry too much, it happens to everyone.

The culprit was yeast. They were cross contaminated in almost all my cells and I must now wait another 2 weeks for the frozen cells to thaw out and grow to be ready for use... I cling to EtOH in the TC room and use it frequently which makes me wonder why I would get contamination.. but oh well.. all i can do is to just focus on being really mindful of the hood and sterile conditions and hope it doesnt happen again!!! lol
Some of the comments here were pretty entertaining tbh. 🙂

 

[crimsonkid85]

The culprit was yeast. They were cross contaminated in almost all my cells and I must now wait another 2 weeks for the frozen cells to thaw out and grow to be ready for use... I cling to EtOH in the TC room and use it frequently which makes me wonder why I would get contamination.. but oh well.. all i can do is to just focus on being really mindful of the hood and sterile conditions and hope it doesnt happen again!!! lol
Some of the comments here were pretty entertaining tbh. 🙂

yeast!!!!!!!!!!!!!!! UV!!!!!!!!!

 

[PreciousHamburgers]

They're just cells...take it easy.

Say this to a militant anti-abortion activist.

 

[mehc012]

Say this to a militant anti-abortion activist.

No thanks, I have several other choice phrases I'd prefer in that situation.

 

[Steven1991]

The culprit was yeast. They were cross contaminated in almost all my cells and I must now wait another 2 weeks for the frozen cells to thaw out and grow to be ready for use... I cling to EtOH in the TC room and use it frequently which makes me wonder why I would get contamination.. but oh well.. all i can do is to just focus on being really mindful of the hood and sterile conditions and hope it doesnt happen again!!! lol
Some of the comments here were pretty entertaining tbh. 🙂

I've heard way too many stories of people who bake bread/make beer having cell cultures contaminated by yeast. Just something to be mindful of 😛

 

[mcloaf]

The culprit was yeast. They were cross contaminated in almost all my cells and I must now wait another 2 weeks for the frozen cells to thaw out and grow to be ready for use... I cling to EtOH in the TC room and use it frequently which makes me wonder why I would get contamination.. but oh well.. all i can do is to just focus on being really mindful of the hood and sterile conditions and hope it doesnt happen again!!! lol
Some of the comments here were pretty entertaining tbh. 🙂

...drosophila?

 

[Patros]

Hm, how do I deal with a contaminated cell line? Dump some bleach in it, move on.

Did you say it takes two weeks to thaw out fresh cells? Are these frozen in the South Pole or something?

 

[Mad Jack]

I don't really understand how you feel myself, but @Dave89 is always complaining about culture contamination. Maybe he's got some advice in dealing with it. Personally, I think it's all in both of your heads and you need to chill.

 

[TheShaker]

Listen buddy, the allied forces bombed the CRAP outta Germany not for you to be wasting cells. YOU COULD OF USED THEM U GOTTA BE KIDDING ME, THE VETERANS WHO SAVED THE ALLIED FORCES FOUGHT SO THAT EVERY MAN COULD HAVE AN EDUCATION AND HERE WE HAVE SOME UNGRATEFUL KID MESSING UP CELL CULTURES. THINK ABOUT HOW THE CELLS FEEL, THEY DIDNT DO ANYTHING WRONG BUT NOW THEY DIE. GOD DAMN IT YOU JUST killed 100M+ CELLS AND U WANNA BE A DOCTOR. DAMN MAN DAMN IT.

What if they were tumor cells? OP has literally cured cancer!!

 

[Mad Jack]

I don't see any connection to Vietnam, Walter

Really glad you brought back the Rob Ford, Reckoner. It just... works for you. You rock it well.

 

["The Stig"]

Hi all,
For those who culture cells for research, if you have ever had contamination in your cells, how do you cope with it? I had some pretty bad cross contamination recently and a month's work of cell work has been lost and I will now have to start over. My mentor already knows about it (she is pretty laid back-and said its okay). Research means a lot to me (something I look forward to-although feeding cells daily can be tiresome). I just feel empty and discouraged. I've had contamination before at a different lab, seen others get it as well but for some reason, I'm just very disappointed in myself and having a hard time finding the energy to pick up from here. :'(

Cry when you work with exclusive tier3 cultures that cost $10,000 the batch.

Unlike some members have commented:

• Do not throw away the contaminated plate
• Do not pour bleach on the monolayer, etc...

It is very tedious for us, mentors, to retrace the source of contamination once you dispose of the evidence.
In the future:
*Conserve the plate, and show it to someone more knowledgeable - Then you can throw it away.*

 

[Mad Jack]

Cry when you work with exclusive tier3 cultures that cost $10,000 the batch.

Unlike some members have commented:

• Do not throw away the contaminated plate
• Do not pour bleach on the monolayer, etc...

It is very tedious for us, mentors, to retrace the source of contamination once you dispose of the evidence.
In the future:
*Conserve the plate, and show it to someone more knowledgeable - Then you can throw it away.*

His cultures are pretty obscure, you've probably never heard of them.

 

[Dral]

Sorry that happened. You seem upset out of proportion though.

Coping mechanisms. I hope you have them if you're going to be a doctor. Heaven forbid a hypothetical where one accidentally forgets to wash hands between patients in the ICU and the second patient picks up MRSA, goes septic and dies. Now THERE are some contaminated/dead cells to be upset about.

 

[Zumab]

Cry when you work with exclusive tier3 cultures that cost $10,000 the batch.

Unlike some members have commented:

• Do not throw away the contaminated plate
• Do not pour bleach on the monolayer, etc...

It is very tedious for us, mentors, to retrace the source of contamination once you dispose of the evidence.
In the future:
*Conserve the plate, and show it to someone more knowledgeable - Then you can throw it away.*

I think the cause was the media that I was using smh :/
Yes, these were all generated from Tier3 cells. Lots of labor, time, resources etc
Oh, well.... gotta move on i guess! lol

 

[Dral]

A few weeks from now, it won't seem like a big deal at all! Just do your best to keep that in mind.

 

[Goro]

You throw them away and thaw out another vial of cells.

Note the word:

Research

Hi all,
For those who culture cells for research, if you have ever had contamination in your cells, how do you cope with it? I had some pretty bad cross contamination recently and a month's work of cell work has been lost and I will now have to start over. My mentor already knows about it (she is pretty laid back-and said its okay). Research means a lot to me (something I look forward to-although feeding cells daily can be tiresome). I just feel empty and discouraged. I've had contamination before at a different lab, seen others get it as well but for some reason, I'm just very disappointed in myself and having a hard time finding the energy to pick up from here. :'(