We are performing Single Cell Colony Formation Assay of MCF-7 cells in our Dissertation Project work.We found the colony after 20 days of experiment. We want to separate out the colony from the agar plate and check the expression of TGF-beta in it.
So please give me any protocol for colony pick up and better method for that.
Simplest way is that you can use a pipette tip to transfer the colony from agar plates. Depending on the size of the colony you may use a 200uL or a 1.0 mL tip. Before picking the colony, cut the tip of the pipette tip (wide enough to pick the colony) and just punch the colony out of the the agar and transfer the colony into a 24 well plate by pipeting medium up and down.
Ok sir.. Thank u..
How can we fix the colony on slide? Can we stored them for one week ?
I would pick the colony up like praveen said and fix it on a slide in mounting medium. You can easily store it for a week in the fridge. Preferably for colonies I use a non-hardset mounting medium. However if you want to stain the colony you could do it in a 96 well plate well or even on the glass cover slip, but you'll have to be very careful not to lose the colony and wash with a very fine pipette tip.
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