Red diode laser induced fluorescence detection with a confocal microscope on a microchip for capillary electrophoresis
| Autor: | D. J. Harrison, Guifeng Jiang, Said Attiya, William E. Lee, Gregor Ocvirk |
|---|---|
| Rok vydání: | 2000 |
| Předmět: |
Detection limit
Microscopy Confocal Microscope Laser diode Chemistry Lasers Excitation filter Biomedical Engineering Biophysics Analytical chemistry Electrophoresis Capillary General Medicine Laser Fluorescence law.invention Capillary electrophoresis law Electrochemistry Dichroic filter Laser-induced fluorescence Biotechnology |
| Zdroj: | Biosensors and Bioelectronics. 14:861-869 |
| ISSN: | 0956-5663 |
| DOI: | 10.1016/s0956-5663(99)00056-1 |
| Popis: | A highly sensitive laser induced fluorescence (LIF) detection system based on a 635 nm laser diode and cyanine-5 (Cy-5) dye, is described for use with a planar, microfluidic, capillary electrophoresis (CE) chip. The CE-chip is able to determine a protein biological threat agent simulant, ovalbumin (Ov), by performing an immunoassay separation of Cy-5 labeled anti-ovalbumin from its complex with Ov, in under 30 s. A confocal, epiluminescent detection system utilizing a photomultiplier tube gave optimum results with a 400 microm pinhole, an Omega 682DF22 emission filter, a 645DRLP02 dichroic mirror, a 634.54 +/- 5 nm excitation filter, and a Power Technology ACMO8 635 nm laser operated at 11.2 mW. Using this detector, a microchip CE device with a separation efficiency of 42,000 plates and an etch depth of 20 microm, gave a concentration detection limit of 9 pM Cy-5. This limit corresponds to the determination of 4560 injected molecules and detection of 900 of these molecules, given a probe volume of 1.6 pl and a probing efficiency of 20%. |
| Databáze: | OpenAIRE |
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