We have an ELISA plate reader in our lab having an option path length correction. I'm just wondering when we should use this function and how it works.
Hi Kumar,
The main function of microplate pathlength correction is to normalizes absorbance values measured on a microplate to correspond with the absorbance values measured in a standard curvette. According to Beer Lambert law, A=E*C*d (where E= Extinction coefficient,C=concentration, d=pathlength(cm)) so, the liquid pathlength in a standard curvette is 1cm whereas the liquid pathlenght on a microplate is not fixed. Microplate pathlength correction is required for calculating sample concentration directly from absorbance value since the absorbance is measured vertically through the well,several factors affect the liquid pathlength, and thus the absorbance.
I hope this will help you.
Hi Kumar,
The main function of microplate pathlength correction is to normalizes absorbance values measured on a microplate to correspond with the absorbance values measured in a standard curvette. According to Beer Lambert law, A=E*C*d (where E= Extinction coefficient,C=concentration, d=pathlength(cm)) so, the liquid pathlength in a standard curvette is 1cm whereas the liquid pathlenght on a microplate is not fixed. Microplate pathlength correction is required for calculating sample concentration directly from absorbance value since the absorbance is measured vertically through the well,several factors affect the liquid pathlength, and thus the absorbance.
I hope this will help you.
Hi Kuma, I totally agree with Adesola. Beer-Lambert law states that the path length has to be 1 cm to be considered Absorbtion.
Anything else is optical density (OD)
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