5 replies to this topic

[ah6tyfour]

I've worked in many labs and have done cell culture for more than 10 years now and I've never seen anyone insist that anything entering the incubator must be sprayed with ethanol and wiped down.  I have just joined a new lab to train the staff and realized the PI has very limited experience in the lab.  

Does anyone think that wiping down their flasks prior to incubating does anything beneficial?  When I first stepped into this lab, the first thing I saw was three researchers wiping down over 20 T-75 flasks one at a time (and dropping one onto the floor).

Ethanol doesn't really kill anything nasty, you run the risk of getting it in your cells, and wiping every surface with a kimwipe is probably just introducing more contaminants.

Am I right in thinking this?

Since I'm hired to get this lab going, I want to put an end to this tedious task, but I want to have solid reasons so I can go to the PI and make my case.  If anyone knows of a paper or a tech note or something that says not to wipe down prior to incubation, I'd really appreciate it.

Edited by ah6tyfour, 23 June 2012 - 03:08 PM.

[Natrium]

We spray everything with 70% ethanol before putting it in the incubator. Same for putting anything in the hood. We don't wipe anything down. I'm not sure of any literature supporting this, but it is "tradition" in our lab. There are several videos on youtube that do this as well.

[bob1]

To my mind there is very little point in spraying/wiping with ethanol before putting in the incubator - it is open to the air and a lot of air gets taken into the incubator each time you open the door. Besides which the flasks/plates are closed and shouldn't pick up too much stuff.

When putting into the hood, yes wipe then, that is when the flasks/plates will be opened.

[leelee]

What bob said

I never spray or wipe anything going into the incubator.

I do, however, always handle my flasks and plates with clean hands (usually gloved and sprayed with ethanol) and I don't typically put them down anywhere except inside the hood or on the microscope.

I actually don't even spray them or wipe them down with ethanol before putting them in the hood (I hate the way it makes my labelling run and come off).  I don't use antibiotics in my media, and I don't get contamination- so its been working fine for me

[ah6tyfour]

Thanks everyone!  I also have never sprayed or wiped down anything going into or coming out of the incubator.  I've never used antibiotics and have never had a problem.  The lab I'm consulting for now does this paranoid wipe down and they lace their media with antibiotics and fungicide (and carry out their actual experiments with these additives in the media).

And leelee, if you use lab markers for labeling, you won't have to worry about your writing running or coming off.  They are resistant to most solvents including ethanol.  These are the only markers I allow in the lab.  I use the fine-tip for tissue culture and conical tubes and other big things.  For labeling microfuge tubes, I use the extra-fine-tip.  Black and blue are the only colors worth buying.  The red and green are a bit strange and hard to read.
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[rhombus]

Dear All,


The reason why you should always spray flasks/plates in and out of the CO2 Incubator is that both areas can have possible contamination risks.

When splitting cells in the Class II Cabinet spills can take place and Media Aerosols can be a major problem. If you do not wipe your flasks/plates then media can be left on the outside of the vessel giving a perfect opportunity for fungal contamination. 99% of all fungal infections in CO2 Incubators are from NOT wiping down your vessels. If you are spliiting cells and then visualising them on a microscope stage before putting them in the Incubator......this is a source of potential infection.....and not wiping them again can compound the problem.

Normally when using a Class II cabinet, the first thing you do is ??????

Wipe it down with 70% IMS (Ethanol)

When I use my pipettor in the cabinet, the first thing I do is ????????????

Wipe it down with 70% (Ethanol)

Gilson's used in the cabinet

Wiped down with 70% IMS (Ethanol)


Etc Etc Etc



This is the way I was taught in the 1970's and have not changed this aspect of my aseptic technique.......seems to work for me..........and no I do not use and do not recommend using antibiotics OR antifungals at all in cell culture (posted many times on the subject).

Kindest regards

Uncle Rhombus