An economical fluorescence detector for lab-on-a-chip devices with a light emitting photodiode and a low-cost avalanche photodiode

Abstract

An economical fluorescence detector was developed with an LED as the exciting source and a low-cost avalanche photodiode (APD) module as a photon sensor. The detector was arranged in an epifluorescence configuration using a microscope objective (20× or 40×) and a dichroic mirror. The low-cost APD was biased by a direct current (DC) high voltage power supply at 121 V, which is much lower than that normally used for a PMT. Both DC and square wave (SW) supplies were used to power the LED and different data treatment protocols, such as simple average for DC mode, software based lock-in amplification and time specific average for SW mode, were tested to maximize the signal-to-noise ratio. Using an LED at a DC mode with simple data averaging, a limit of detection of 0.2 nmol L⁻¹ for sodium fluorescein was attained, which is among the lowest ever achieved with an LED as an excitation source. The detector was successfully used in both capillary and chip electrophoresis. The most significant advantages of the detector are the compact size and low cost of its parts. The aim of the work is to prove that widely available, low-cost components for civilian use can be successfully used for miniaturized analytical devices.