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A microplate reader uses colorimetric, fluorometric, or luminescent reagents that measure the concentration of the compound of interest by developing a signal that can be detected by the reader, similar to a traditional spectrophotometer but able to process 96, 394 or 1536 samples in one read sequence – quickly!

The light detector is located on the opposite side of the microplate relative to the light source. It measures how much of the total light intensity is transmitted through the probe and at which wavelengths. Light that does not pass through to the detector is absorbed.

Working principle of a microplate reader A microplate reader detects light signals produced within a specific wavelength range by samples that have been pipetted into a plate. The optical properties of these samples are the result of a biological, chemical, biochemical, or physical reaction.

Fluorescence microplate readers usually contain a light source, an optical system for the selection of excitation light (i.e., filters and/or monochromators), a second optical system for the selection of emission light, and a detector, usually one or multiple photomultiplier tubes (PMTs).

A spectrophotometer offers higher dynamic range and precision and is less sensitive to inconsistencies in sample sizes, but a microplate reader supports more samples at once, making it crucial for high-throughput operations.