ELISA Reader Instrumentation Quiz

ELISA Reader Function and Principle

• The primary function of an ELISA reader is to detect and quantify substances such as nucleic acids and carbohydrates.
• An ELISA reader is based on the principle of absorbance, which is measured using the Beer-Lambert law.

Instruments Combined in an ELISA Reader

• An ELISA reader combines a fluorimeter and a spectrophotometer (not a mass spectrometer).

Beer-Lambert Law and Absorbance

• The Beer-Lambert law states that the absorbance of the solution has a linear relationship with the concentration of the sample.
• The law is expressed as A = εcl, where A is the absorbance, ε is the molar absorptivity, c is the concentration of the sample, and l is the path length.
• The fraction of light passing through the sample is expressed as T = It/Io, where T is the transmittance, It is the intensity of the transmitted light, and Io is the initial intensity of the light.

ELISA Technique and Detection

• The ELISA technique is designed for detecting and quantifying substances such as nucleic acids and carbohydrates.
• The ELISA reader can detect and quantify these substances in a 96-well plate.

Note

• An ELISA reader does not read the fluorescent response of the ELISA, as it is a spectrophotometer that measures absorbance, not fluorescence.