Image Scanning Microscopy (ISM)

In Image Scanning Microscopy (ISM), fluorescence is detected using a multi-element detector array instead of a single pinhole detector. Each detector element effectively functions as a miniature confocal pinhole, maintaining strong optical sectioning. During image reconstruction, detected signals are computationally shifted toward their true emission origin through pixel reassignment, which enhances spatial resolution beyond conventional confocal imaging. Further improvements in contrast and detail can be achieved with focus-variant ISM and optional deconvolution algorithms.

NovaISM is dedicated software for the reconstruction and analysis of Image Scanning Microscopy data. It integrates pixel reassignment, computational sectioning, and deconvolution to enhance spatial resolution and optical sectioning in a unified workflow.

Robust Image Scanning Microscopy measurements require confocal laser scanning combined with spatially resolved photon detection and precise data acquisition. Key instrumentation requirements include:

• Confocal scanning microscope equipped with a detector array, such as a SPAD array, to capture spatially resolved fluorescence signals.
• Multi-element photon-counting detector arrays, such as SPAD or multi-anode PMT arrays, with low noise and high sensitivity to ensure accurate spatial photon detection and signal integrity across channels.
• Dedicated reconstruction and analysis software to perform pixel reassignment, computational sectioning, and optional deconvolution for resolution and contrast enhancement.

Together, these components provide resolution-enhanced confocal imaging while maintaining optical sectioning capability and quantitative fluorescence contrast.