I am measuring the absorbance of a Methylene Blue gradient on a plate reader.
The highest concentrations are "super dark blue" in colour and on the calibration plot begin to plateau (no longer linear)
If I dilute the "super dark blue" concentrations by 50% prior to plate reading the absorbance is this an accurate method?
(I tried this for one point and its absorbance then fell linearly onto my calibration graph...but the undiluted "super dark blue" was a lower absorbance than the calibration trendline).
So wanted to check if this was a coincidence. Or whether this is scientifically valid.
Thanks.
Dear Theatin
- For my opinion, to be in the safe zone, try to downgrade the concentrations of your serial dilution. That means make sure also there is no sample concentration out of your calibration graph concentrations.
Thanks Hazem. I am going to keep the calibration within the working range of the dye.
Then for my samples will do a test run and see what requires dilution and then dilute the samples first before adding the dye to also keep them in the working range.
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