Image Scanning Microscopy

Discover next-generation confocal microscopy

Image Scanning Microscopy (ISM) can improve the effective spatial resolution of conventional confocal laser scanning microscopy to its theoretical limit, without compromising any of its functionalities.

Our team implemented a natural design of ISM based on a fast single-photon detector array which allows straightforward upgrade of confocal microscopy and provides access to the raw scanned images, also paving the way to adaptive reconstruction methods.

While standard all-optical implementations of ISM can reduce versatility, our solution enables for a direct combination with fluorescence spectroscopy techniques, such as fluorescence lifetime imaging and fluorescence correlation spectroscopy.


Push confocal microscopy resolution to its theoretical limit by achieving a 2-fold improvement.
PRISM application 1


Achieve the same resolution as ideal confocal at one-tenth the illumination intensity.
PRISM application 2


Collect photons in time-tag modality in order to perform super-resolution time-resolved imaging.
PRISM application 3

Read more about our work

Selected publications
fiber_new Focus-ISM for Sharp and Gentle Super-Resolved Microscopy.
G. Tortarolo et al.
bioRxiv (2022).
fiber_new ISM-FLUX: single-step MINFLUX with an array detector.
E. Slenders et al.
bioRxiv (2022).
Confocal-based fluorescence fluctuation spectroscopy with a SPAD array detector.
E. Slenders et al.
Light: Sci. Appl. (2021).
Cooled SPAD array detector for low light-dose fluorescence laser scanning microscopy.
E. Slenders et al.
Biophys. Rep., 557-567 (2021).
The BrightEyes-TTM: an Open-Source Time-Tagging Module for Single-Photon Microscopy.
A. Rossetta et al.
bioRxiv (2021).
SPAD-based asynchronous-readout array detectors for image-scanning microscopy.
M. Buttafava et al.
Optica 7, 755-765 (2020).
Pixel reassignment in image scanning microscopy: a re-evaluation.
C. J. R. Sheppard et al.
J. Opt. Soc. Am. A 37, 154-162 (2020).
Two-photon image-scanning microscopy with SPAD array and blind image reconstruction.
S. V. Koho et al.
Biomed. Opt. Express 11, 2905-2924 (2020).
A robust and versatile platform for image scanning microscopy enabling super-resolution FLIM.
M. Castello et al.
Nat. Methods, vol. 16, no. 2, pp. 175-178 (2019).
Smart scanning for low-illumination and fast RESOLFT nanoscopy in vivo.
J. Dreier et al.
Nat. Commun., vol. 10, no. 1, p. 556 (2019).
Image formation in image scanning microscopy including the case of two-photon excitation.
C. J. R. Sheppard et al.
J. Opt. Soc. Am. A, vol. 34, no. 8, pp. 1339-1350 (2017).
Image scanning microscopy with a quadrant detector.
M. Castello et al.
Opt. Lett., vol. 40, no. 22, p. 5355 (2015).
Multi-images deconvolution improves signal-to-noise ratio on gated stimulated emission depletion microscopy.
M. Castello et al.
Appl. Phys. Lett., vol. 105, no. 23, p. 234106 (2014).