Highly Sensitive Cy3 Fluorescence Detection Method for Microarray Based on CCD and LED.

Microarrays play a crucial role in detecting diverse biological assays, supported by high-throughput fluorescent scanners that stem from laser confocal scanning mechanisms or charge coupled device (CCD) wide-field imaging technology. Nonetheless, each scanner exhibits significant limitations. To address these drawbacks, this study proposes a novel design for detecting cyanine-3 (Cy3) fluorescence microarrays using a cooled CCD and high-power light emitting diode (LED), which demonstrates excellent performance to overcome the high complexity of the laser confocal technique and the low efficiency of high-pressure gas lamps in traditional CCD scanner by ExviewHAD CCD of ICX694AL and single-core powerful LED of PT54G. The experimental results, including comparative testing of BAC Microarray of MGmed Inc., Republic of Korea (MGMED BAC) microarrays using laser confocal scanning and cooled CCD reading, reveal that the novel CCD scanner achieves a detection sensitivity of 1.12 fluor/mu m(2), a reading resolution of 10.0 mu m, and a measuring repeatability indicator R-v <= 2.33%. In addition, to mitigate no-linear response in quantitative detection of Cy3 fluorophores, the calibration curve is analyzed by the least-squares approximation (LSA), and the LSA fitting of N=3 in the CCD linearity region yields a maximum relative error of only 2.35%. These exceptional performance metrics indicate that this newly developed CCD wide-field scanner integrated with a high-power LED exhibits excellent capability in detecting Cy3 microarrays.