Dear All,
I am doing experiment to see effects of treating cell culture with different chemical substances. I have to treat the cell with liquid substance A with specific concentration first (pre-treat), then after some hours, treat with liquid substance B liquid (post-treat). When treating with substance B the total volume of media in cell culture would be increase and the final concentration of substance A would change. Is there any limit of percentage change in final concentration that is acceptable?
Thank you very much,
Well I don't know the answer to the question regarding the volume you can add but here are my thoughts about your problem: when you add substance A during the few hours (before B) the concentration of this substance is changed! It probably reacts with different cell constituents and structures. Especially if the cell line is metabolically active where your substance is metabolized, so the starting concentration is most certainly decreased. In order to avoid to change the conc. of substance A add as low volume of B as posible (like 5uL or less if it is posible).
all best
Hi Asa, Can I asked you if you have problem in changing the medium with a fresh one? In this fresh medium, for example 1ml you can add again substance A and B at the correct concentration; I don't known if this is a good suggestion or you faced other problems with your treatments.....
If the effect of substance A is confined to the time prior to the addition of substance B, then the dilution of substance A does not matter, On the other hand, if the effect of substance A continues into the team following the addition of substance B, then you should minimize as much as possible the dilution of substance A. I would try to keep the dilution to 10% or less. Also, document the amount of dilution when describing the method.
I would say there is no generic answer to this good question. Have you ever made a chocolate mouse from scratch, or attempted recipes that are classics, such as beer or wine from first principles? These types of systems hinge on geometric progressions in parameter spaces that do not have strictly definable outcomes, or properties that scientists typically like to believe they are working with (ie that can be written down). The suggestion i would share is to figure out how to validly measure the properties of the system you are studying that are critical to you. Then, with this metric (assay) in hand, you vary the parameters of the system until you get into a region of linear response for this property. The property you seek to study and measure has to be in the system, if this will work. You can't make a Samurai sword from any clod of dirt. This is conceptually identical to building a nuclear reactor for the first time, since you are only doing good work if no one has done it before. Cheers!
Hey,
why do you not prepare Substance B's stock solution in medium which already has the final concentration of Substance A? Then you can mix the two solutions in your wells in any proportion, which will however not change Substance A's concentration no matter what. So you dont have to make any bad compromise in accepting altered concentrations.
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