Why is the enzyme-linked immunosorbent assay (ELISA) instrument only having a few fixed wavelengths

Enzyme-linked immunosorbent assay (ELISA) is commonly used to measure the results of chemical reactions under absorbance detection, which typically involve interactions between certain substrates and enzymes. In absorbance detection, selecting the appropriate wavelength is crucial, and there are some technical and theoretical reasons why the ELISA reader only has a few fixed wavelengths.

Why is the enzyme-linked immunosorbent assay (ELISA) instrument only having a few fixed wavelengths

Specific absorbance peaks of substrates: Substrates and products exhibit different absorbance characteristics at different wavelengths. Usually, substrates and products absorb light within the UV visible spectral range, and their maximum absorbance points (wavelengths) are fixed. Therefore, selecting a specific fixed wavelength can more accurately measure changes in the reaction.

Specific working wavelength of enzymes: Enzymes typically exhibit high catalytic activity at specific wavelengths. Therefore, selecting an appropriate wavelength for measuring the generation of enzyme catalyzed products can improve sensitivity and accuracy.

Standardization and comparison: Fixed wavelengths help standardize experiments and compare results. When using the same wavelength for measurement, it is easier to compare data between different experiments because they are conducted under the same optical conditions.

Instrument design and cost: In order to provide flexibility and practicality, manufacturers usually choose several fixed wavelengths in the enzyme-linked immunosorbent assay (ELISA) reader, which cover the needs of most common experiments. Adding more wavelength optical components may increase the complexity and cost of the equipment.

Specific application requirements: For specific enzyme activity or reaction properties, selecting a specific wavelength is necessary. In this case, users may need to use higher-level devices with adjustable wavelengths.

Why is the enzyme-linked immunosorbent assay (ELISA) instrument only having a few fixed wavelengths

Overall, a fixed wavelength enzyme-linked immunosorbent assay (ELISA) reader can meet most common experimental needs, while also making instrument design simpler, easier to use, and more economical. If a particular experiment requires the use of very specific wavelengths, it may be necessary to consider using more flexible spectroscopic instruments.