High-speed sCMOS acquisition with optical pixel reassignment

Fluorescence microscopy has undergone rapid advancements yielding unprecedented visualization of biological events and shedding light on the intricate mechanisms governing living organisms. Scientific-grade CMOS (sCMOS) sensors offer high signal-to-noise ratios, quantum efficiency over 90%, and low readout noise under 2e−, making them suitable for low-light imaging [1-4]. However, the trade-off between speed and field of view often limits their full potential, making it challenging to capture rapid biological events below the millisecond scale. Here, we introduce sHAPR, a high-speed acquisition technique that leverages the operating principles of sCMOS cameras to capture fast cellular and subcellular processes. The system utilizes a custom fiber bundle to compress a 2D image into a linear output, enabling ultra-high-speed recording (>25kHz). Attached to an epifluorescence microscope, the sHAPR system demonstrated its potential in applications such as high-throughput flow cytometry, cardiomyocyte contraction imaging, and neuronal calcium wave tracking.