Tips for ELISA Reader Operation

An ELISA analyzer is a specialized spectrophotometer designed to read the results of this technique, which is used to determine the presence of antibodies or specific antigens present in a sample. The technique is based on the detection of an antigen immobilized on a solid phase, by means of antibodies that, directly or indirectly, produce a reaction whose product can be read by the spectrophotometer. It is also known as ELISA reader.

What is the principle of its operation?

As mentioned above, the ELISA analyzer is a specialized spectrophotometer. Unlike conventional spectrophotometers that allow readings over a wide range of wavelengths, this one has diffraction filters or gratings that limit the range of wavelengths to those used in the ELISA technique, which is generally performed with wavelengths between 400 and 750 nm -nanometers-. Some analyzers operate in the ultraviolet range and can perform analyses between 340 and 700 nm.

What equipment is required to perform ELISA assays?

To develop the ELISA technique, at least the following equipment is required:

⦁ An ELISA analyzer.
⦁ An ELISA washer.
⦁ A liquid dispensing system (multichannel pipettes can be used).
⦁ A specialized incubator for the plates.

What is calibration like?

The calibration of an ELISA analyzer is a specialized process that must be performed by a properly trained technician or engineer, following the instructions provided by each manufacturer. To perform the calibration it is necessary to have a set of gray filters, which are mounted on a plate of the same geometry as those used to perform the analysis. These filters are supplied by the manufacturers and can be used to perform calibrations at any wavelength used by the equipment.

What service does this equipment require?

For the ELISA analyzer to operate correctly, the following points need to be verified:

⦁ A clean, dust-free environment.
⦁ A stable work table. It is advisable that it is far away from equipment that generates vibrations -centrifuges, shakers-, and that it has an adequate size to place, next to the ELISA analyzer, the complementary equipment required to perform the technique in question: washers, incubator, dispenser and computer with peripherals.
⦁ A power supply according to the norms and standards implemented in the country. In American countries, voltages of 110 V and frequencies of 60 Hz are generally used.

What should be the maintenance routine?

Basic maintenance (Frequency: Daily)

⦁ Check that the optical sensors of each channel are clean. If dirt is detected, clean the surface of the light emitter windows and sensors with a brush.
⦁ Confirm that the lighting system is clean.
⦁ Verify that the analyzer calibration is adequate. When starting daily operations, allow the analyzer to warm up for 30 minutes. Next, take a blank reading and then read a module full of substrate. The readings should be identical. If they are not, invert the module and repeat the reading in order to determine if the deviation originates from the module or the reader.
⦁ Examine the automatic plate feed. It should be smooth and constant.
Preventive Maintenance (Frequency: Quarterly)
⦁ Check the stability of the lamp. Use the calibration filter, taking readings at 30-minute intervals or the same plate. Compare readings. There should be no differences.
⦁ Clean the optical systems of the detectors and the illumination systems.
⦁ Clean the plate feed mechanism.
⦁ Verify the alignment of each well with the light emitting systems and detectors.

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