Two-photon image-scanning microscopy with SPAD array and blind image reconstruction
Eli Slenders; Marco Castello; Alberto Diaspro; Elena Tcarenkova; Mauro Buttafava; Federica Villa; Marcel Ameloot; Alberto Tosi; Paolo Bianchini; Giuseppe Vicidomini; Sami V. Koho; Colin J. R. Sheppard; Giorgio Tortarolo
Two-photon image-scanning microscopy with SPAD array and blind image reconstruction
Eli Slenders
Marco Castello
Alberto Diaspro
Elena Tcarenkova
Mauro Buttafava
Federica Villa
Marcel Ameloot
Alberto Tosi
Paolo Bianchini
Giuseppe Vicidomini
Sami V. Koho
Colin J. R. Sheppard
Giorgio Tortarolo
OPTICAL SOC AMER
Julkaisun pysyvä osoite on:
https://urn.fi/URN:NBN:fi-fe2021042823854
https://urn.fi/URN:NBN:fi-fe2021042823854
Tiivistelmä
Two-photon excitation (2PE) laser scanning microscopy is the imaging modality of choice when one desires to work with thick biological samples. However, its spatial resolution is poor, below confocal laser scanning microscopy. Here, we propose a straightforward implementation of 2PE image scanning microscopy (2PE-ISM) that, by leveraging our recently introduced single-photon avalanche diode (SPAD) array detector and a novel blind image reconstruction method, is shown to enhance the effective resolution, as well as the overall image quality of 2PE microscopy. With our adaptive pixel reassignment procedure similar to 1.6 times resolution increase is maintained deep into thick semi-transparent samples. The integration of Fourier ring correlation based semi-blind deconvolution is shown to further enhance the effective resolution by a factor of similar to 2 - and automatic background correction is shown to boost the image quality especially in noisy images. Most importantly, our 2PE-ISM implementation requires no calibration measurements or other input from the user, which is an important aspect in terms of day-to-day usability of the technique. (C) 2020 Optical Society of America under the terms of the OSA Open Access Publishing Agreement
Kokoelmat
- Rinnakkaistallenteet [19207]