Thin-film optical transducers integrated in lab-on-a-chip (LoC) devices have the potential to facilitate miniaturization, multiplexing capabilities, portability, and sensitivity. However, there are few systematic studies that provide detailed characterization of these photosensors to particular miniaturized bioassays at reduced light intensities. A more detailed representation of the photosensors performance at low light levels is thus needed in order to improve the engineering of integrated detectors for the next generation of portable biosensors. Here, an assessment of the performance of hydrogenated amorphous silicon (a-Si:H) photosensors, at ultra-low light intensity, based on two different device configurations for integration in a microfluidic platform for biomolecular detection is presented. Both p-i-n photodiodes and parallel contact photoconductors show low dark current density (~10−10 A.cm−2) and photosensitivity comparable with high performing crystalline silicon photosensors. The a-Si:H photosensors were integrated in microfluidic devices for the detection of antibody-horseradish peroxidase conjugates (Ab-HRP) in a chemiluminescence-based assay. Surface concentrations of Ab-HRP as low as ~26 amol.cm−2 and ~9 fmol.cm−2 (corresponding to ~103 and ~106 HRP molecules) were detected with detectors in photodiode and photoconductor configurations, respectively, with the sensitivity of the former being comparable with that of bulky and ultra-sensitive photomultiplier tubes. The excellent sensitivity and dynamic range obtained, together with the miniaturization and arraying potential, highlight the potential of a-Si:H photosensors as an effective means of integrating the multiplexed acquisition of optical data in real time in point-of-need LoC systems.