High dynamic range shortwave infrared imaging of mice with an InGaAs camera.

Significance: Although shortwave infrared (SWIR) imaging provides superior tissue penetration and reduced autofluorescence for preclinical applications, quantitative fluorescence analysis is hindered by the limited dynamic range (DR) of InGaAs cameras, forcing a focus on either bright or dim anatomical features. Aim: We develop a high dynamic range (HDR) imaging method specifically adapted for the high-noise characteristics of InGaAs detectors to enable quantitative fluorescence imaging across wide intensity ranges. We demonstrate that one-time camera calibration based on a series of images encompassing the range of radiance intensities enables all subsequent image processing. Approach: We modified classical HDR algorithms with exposure-time-dependent dark current subtraction, preprocessing to exclude saturated and noisy pixels before camera response function recovery, and dynamic weighting range adjustment to account for shrinking intensity ranges at longer exposures. High dynamic range image processing effects on preclinical imaging outcomes were analyzed using indocyanine green and SWIR-emitting PbS/CdS quantum dots in mouse models. Results: High dynamic range imaging achieved a 22-dB improvement in DR over single exposures, enabling simultaneous quantification across more than three orders of magnitude of fluorophore concentration. Conclusions: This software-based HDR SWIR imaging approach eliminates exposure parameter optimization and enables comprehensive biodistribution analysis across all anatomical structures from a single acquisition sequence, significantly streamlining preclinical imaging workflows while preserving quantitative accuracy.