Microplate Reader

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Post on 13-May-2015

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Microplate reader is a device which used to read the result of ELISA test utilizing theory of spectrophotometer.

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<ul><li>1.Microplate Reader It is a device which used to read the result of ELISA testutilizing theory of spectrophotometer. ELISA-(Enzyme-Linked Immunosorbent Assay) Biochemical technique used mainly in Immunology to detect the presence of an antibody or an antigen in a sample. Also called Photometric Microplate reader, ELISA reader</li></ul> <p>2. Theory of operation Microplate reader is specialized spectrophotometer which facilitatesreadings on a wide range of wavelengths, generally between 400 to 750 nm. Beer Lambert law, A = absorption = absorption coefficient l = path length c = concentration I0 = reference light intensity I = light intensitycl A light beam, passing through the sample has a diameter ranging between1 to 3 mm. Then detection system detects the light coming from the sample, amplifies the signal and determines the samples absorbance. 3. Theory of operation 4. Internal Parts Light SourceELISA PlatePower supply 5. Installation A clean, dust free environment. A stable work table away from equipment that vibrates(centrifuges, agitators). 6. calibration Calibration plates are equipped with at least threepredetermined optic density values within the measurement ranges; low, medium, and high value. Place the calibration plate on the equipment and carry out a complete reading. Verify if there are differences in the readings obtained from well to well. If this is the case, invert the plate (180) and repeat the reading to rule out that differences are attributed to the plate itself. Identify the reader requires calibration. If so, proceed with the calibration according to manufacturer. 7. Maintenance Basic : DailyPreventive : QuarterlyReview that optical sensors of each channel are clean. 2. If dirt is detected, clean the light emitters and the sensors with a small brush. 3. When the daily operations begin, let the reader warm up for 30 minutes. Next, do a blank reading and then read a full plate of substrate. The readings must be identical. 4. Examine the automatic drawer sliding system. It must be smooth and constant.1.1.2. 3. 4.Verify the stability of the lamp. Use the calibration plate conducting readings with intervals of 30 minutes with the same plate. Compare readings. There must be no differences. Clean the detectors optical systems and the lighting systems. Clean the plate drawer. Verify the alignment of each well with the light emission and detection systems. 8. Basic fault Identification The reader gives a reading that does not make sense 2. The readers readings vary from row to row. 3. The reader displays unexpected variation in the optical density readings. 1.4. The reader displays a gradual increase or decrease from column to column.5. Low reproducibility. 6. Misaligned light beam. 9. 1.0 The reader gives a reading that does not make sense ? The illumination lamp is out of service. Replace the lamp with one with thesame characteristics as the original. 10. 2.0 The readers readings vary from row to row. Dirty opticalsensors. The illumination systems lenses or parts are dirty. Lack of calibration in one or more channels. Clean the sensors. Clean the lighting systems lenses. Verify the calibration of each one ofthe channels. 11. 3.0 The reader displays unexpected variation with different wavelengths. The readers lamp is unstable. Replace the lamp with one that hassimilar characteristics as the original. 12. 4.0 The reader displays a gradual increase or decrease from column to column. Inappropriate calibration of the plates controlling motor. Calibrate the plates controllingsystem so that at each step the wells remain exactly aligned with the lighting system. 13. 5.0 Low reproducibility. Sample homogeneity. Mix the reagents before use. Allow these toequilibrate to room temperature. Incorrect pipetting procedure. Ensure pipettes tips are changed between samples and that excessive liquid inside is removed Check the calibration. Reader not calibrated. Reading without sufficient warm up time. Expired reagents. Insufficient or Inefficient washing Wait until the reader has warmed up to its operating temperature. 14. 6.0 Misaligned light beam. The reader was transferred or moved without using Call the specialized service technician.the necessary precautions.The light source has been changed and thereplacement has not been installed or aligned incorrectly Align according to manufacturer instructions 15. Thank you. </p>

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