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mtt at early time points?


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5 replies to this topic

#1 outremer

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Posted 25 January 2011 - 04:20 AM

Hi, Im new to this field. Is it a good idea to quantify amount of adherent cells on titanium at early time-points(30 min, 1h ...) with MTT assay?

One thought I had was that newly trypsinized cells that are seeded on a surface are not very happy, which perhaps affect their metabolic activity - measured by MTT.

So my question, is MTT a suitable technique for newly trypsinized cells adhered on a surface for 30 min?

Grateful for any help!

/Erik

#2 genehunter

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Posted 25 January 2011 - 08:59 AM

If you want to quantify how many cells attached to the surface and compare to late time points after the removal of unattached cells, yes, you could do that.

Remember that this assay takes 2- 4 hrs of incubation time to allow the bioconversion of MTT into colored product.

#3 outremer

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Posted 25 January 2011 - 11:56 PM

If you want to quantify how many cells attached to the surface and compare to late time points after the removal of unattached cells, yes, you could do that.

Remember that this assay takes 2- 4 hrs of incubation time to allow the bioconversion of MTT into colored product.


okey nice, so you dont think that there is any big difference in amount of metabolic activity per cell 15 vs 180 minutes after seeding?

Erik

#4 genehunter

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Posted 09 February 2011 - 10:19 AM

I dont believe this technique is the right one to use to assess any metabolic changes during cell reattachment, because the time (2-4 hrs) required for cells to convert the substrate.

#5 outremer

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Posted 28 February 2011 - 04:49 AM

okey, thanks for the input. I think Ill quantify with DAPI instead...

Erik

#6 dw237606

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Posted 28 February 2011 - 12:25 PM

I plate my cells 12-24 hours before changing the media out and adding my various treatments.




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