Frequently Asked Questions

Microplate Instrumentation

Question

Why do I get different readings from the same sample in different wells in the mictrotiter plate?

Answer

There are many possible sources of assay imprecision. Among these, the quality of the microplate (microtiter plate) is paramount to the quality of the measurements because a microplate reader sends light through a sample vertically, either from top to bottom or bottom to top. Use of a high quality, optically clear microplate is important for reproducibility of sample measurements.

In addition, it is important to use a reference wavelength when measuring a microplate. Microplates may have smudges, scratches, lids and other forms of optical interference which may obstruct the light beam. A reference wavelength allows for a well by well control of imperfections in the plate.

When choosing a reference wavelength, use a wavelength at which the sample of interest does not absorb. 620nm is a frequently used reference wavelength in many absorbance applications in the visible range. Check the product data sheet for recommended reference wavelength for your specific assay.


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