BrightEyes-Scope: A Reference Implementation for Image Scanning Microscopy with SPAD Array Detection
- 1Istituto Italiano di Tecnologia
Abstract
Image scanning microscopy (ISM) is emerging as the natural evolution of confocal laser-scanning microscopy, combining optical sectioning and a two-fold improvement in lateral resolution with high signal-to-noise ratio. Single-photon avalanche diode (SPAD) array detectors enable an effective implementation of ISM by recording a micro-image at each scan position with single-photon sensitivity, negligible readout noise, and without the fixed frame-readout constraints of conventional cameras. Nevertheless, SPAD array-based ISM remains largely confined to laboratories with expertise in microscope development. Here, we present BrightEyes-Scope, a reference implementation of a SPAD array-based ISM system. We provide step-by-step guidelines for optical design, system construction, alignment, calibration, data acquisition, and image reconstruction. Presented as a complete ISM microscope, BrightEyes-Scope also serves as a reference for upgrading existing laser-scanning microscopes. We anticipate that it will accelerate the adoption of ISM for quantitative super-resolution imaging.
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BibTeX
@misc{l2026brighteyesscope,
title = {{BrightEyes-Scope: A Reference Implementation for Image Scanning Microscopy with SPAD Array Detection}},
author = {L. LE and A. Zunino and E. Slenders and A. Bucci and M. Donato and G. Vicidomini},
howpublished = {bioRxiv},
year = {2026},
month = sep,
doi = {10.64898/2026.09.30.755689},
url = {https://www.biorxiv.org/content/10.64898/2026.09.30.755689v1},
note = {CC BY-NC-ND},
}
RIS
TY - UNPB
TI - BrightEyes-Scope: A Reference Implementation for Image Scanning Microscopy with SPAD Array Detection
AU - L. LE
AU - A. Zunino
AU - E. Slenders
AU - A. Bucci
AU - M. Donato
AU - G. Vicidomini
PY - 2026
DA - 2026/09/30
PB - bioRxiv
DO - 10.64898/2026.09.30.755689
UR - https://www.biorxiv.org/content/10.64898/2026.09.30.755689v1
AB - Image scanning microscopy (ISM) is emerging as the natural evolution of confocal laser-scanning microscopy, combining optical sectioning and a two-fold improvement in lateral resolution with high signal-to-noise ratio. Single-photon avalanche diode (SPAD) array detectors enable an effective implementation of ISM by recording a micro-image at each scan position with single-photon sensitivity, negligible readout noise, and without the fixed frame-readout constraints of conventional cameras. Nevertheless, SPAD array-based ISM remains largely confined to laboratories with expertise in microscope development. Here, we present BrightEyes-Scope, a reference implementation of a SPAD array-based ISM system. We provide step-by-step guidelines for optical design, system construction, alignment, calibration, data acquisition, and image reconstruction. Presented as a complete ISM microscope, BrightEyes-Scope also serves as a reference for upgrading existing laser-scanning microscopes. We anticipate that it will accelerate the adoption of ISM for quantitative super-resolution imaging.
ER -