A combination of fluorescence detection and microfluidic technology provides promising applications in life sciences. A prototype of an integrated fluorescence detection system and optical fiber light guide on a laminate-based multichannel microfluidic chip has been developed and tested. A blue LED, plastic optical fiber, photodiode, Mylar and PMMA, and fluorescein and BSA-FITC were used as an excitation source, light coupler and guide, detector, microfluidic substrate and sample, respectively. The results show that the system is capable of detecting weak fluorescence emission from a fluorescein solution at concentration down to 0.01 ng/ml, and gives linear response. The results were also reproducible, and no cross-talk between adjacent channels was observed. The test using BSA as a model analyte demonstrates its feasibility for on-chip immunosensor applications. The performance and applications can be developed further. This prototype can be used as a platform to develop a simple and compact bio-fluorescence detection system integrated with an inexpensive and disposable multichannel microfluidic chip for biomedical devices.