alamar blue - (Jul/13/2009 )

Hi Guys,

Is it possible to do the alamar blue test every day on the same cells for a few days (3) ? Can i still keep the cells after the AB-test and grow them further ?

Ann74 on Jul 13 2009, 03:43 PM said:

I have worked with alamer blue before, but never reused the same cells.

I do wonder why you want to reuse the same cells? You do not use a mircoplate?

Alamar blue has no bad effects on the cells , its non toxic. However: it does change the color of your cells, suspension.
Wich is offcourse the purpose since if the cells grow, you will get a change in color.

Give more details about what you exactly mean with same cells for a few days.

(ex.: you mean you put alamar in your cells in a micro-plate, measure the absorbance , then store this plate and measure again the next day with new alamar? Or?)

One of the things about the alamar blue assay is that the fluorescence intensity increases over time as the cells reduce the dye. You can track this increase over time sure, but at some point the cells will have reduced all of the available alamar in the well and you will start to see decreases in your signal. If you're planning on doing a time course you would basically need one microtiter plate of cells per day of treatment. Alamar blue would be added at the same time each day and incubated for the duration of time that you determined to be optimal for your cells.

And yes you can further culture the cells after the end of the incubation. I've performed clonogenic assays at the end of 5 day treatment periods and have not seen adverse effects of the dye.

hello,

we want to look for the cells during 3 days, see how they grow. We want to see how long you can grow in a 96well plate before you have a 70% confluent layer. We are not satisfy with our results from previous test. that's why we want to try a few things and this is one of them.

And yes, I do use fluorescence to measure my alamar blue test.

Ann74 on Jul 13 2009, 09:30 PM said:

Eum, have you checked the optimum incubation period?

because I do not completely understand your problem or question.

It is possible that you would need 3 days just to measure an optimum result.

Have you allready checked for an optimum incubation period?
(optimizing cell density)

What cells are you growing anyway?

In general: you can prepare your multiwells , store them in the incubator and check the growth every hour or every X hours.. thats no problem at all.
But you need to do to the optimizing cell density check first to have data to compare your new result.


Did you read the AB brochure on how to work with AB ?
In the old manual there is a faq section with some answers to your question.


And cancergreek is right.