2PEF by 15fs laser pulses, 2PEF in image scanning microscopy and 2PE-STED microscopy

Paolo Bianchini; Marco Scotto d'Abbusco; Marco Castello; Giuseppe Vicidomini; Alberto Diaspro

doi:10.1117/12.3001545

13 March 2024 2PEF by 15fs laser pulses, 2PEF in image scanning microscopy and 2PE-STED microscopy

Paolo Bianchini, Marco Scotto d'Abbusco, Marco Castello, Giuseppe Vicidomini, Alberto Diaspro

Proceedings Volume PC12847, Multiphoton Microscopy in the Biomedical Sciences XXIV; PC128470G (2024) https://doi.org/10.1117/12.3001545
Event: SPIE BiOS, 2024, San Francisco, California, United States

Abstract

Two-photon excitation fluorescence microscopy (2PEM) has transformed biological imaging, offering deeper tissue penetration, reduced photodamage, and improved 3D resolution. Here, we explore a cutting-edge 15 fs ultrafast laser for enhanced 2PEM. Our setup includes a single-photon avalanche diode (SPAD) array for image scanning microscopy (ISM) and a 775 nm pulsed laser for STED microscopy. The ultrashort excitation pulse enables higher peak powers and enhanced image clarity due to wider bandwidth. ISM with a 7x7 SPAD array (Genoa Instruments s.r.l., Genoa, Italy) provides crucial insights into imaging conditions, ensuring high-resolution and high-SNR imaging under sub-optimal and deep imaging scenarios. Additionally, our 775 nm pulsed laser implements STED for super-resolution 2PEF imaging beyond the diffraction limit. Integrating these techniques in a single microscope platform unlocks powerful nanoscale investigations in tissues, uncovering complex cellular interactions and paving the way for breakthroughs in neurobiology, developmental biology, and cancer research.

Conference Presentation

Citation Download Citation

Paolo Bianchini, Marco Scotto d'Abbusco, Marco Castello, Giuseppe Vicidomini, and Alberto Diaspro "2PEF by 15fs laser pulses, 2PEF in image scanning microscopy and 2PE-STED microscopy", Proc. SPIE PC12847, Multiphoton Microscopy in the Biomedical Sciences XXIV, PC128470G (13 March 2024); https://doi.org/10.1117/12.3001545

ACCESS THE FULL ARTICLE

INSTITUTIONAL
Select your institution to access the SPIE Digital Library.

SELECT YOUR INSTITUTION

PERSONAL
Sign in with your SPIE account to access your personal subscriptions or to use specific features such as save to my library, sign up for alerts, save searches, etc.

PERSONAL SIGN IN

No SPIE Account? Create one

PURCHASE THIS CONTENT

SUBSCRIBE TO DIGITAL LIBRARY

50 downloads per 1-year subscription

Members: $195

Non-members: $335 ADD TO CART

25 downloads per 1 - year subscription

Members: $145

Non-members: $250 ADD TO CART

PURCHASE SINGLE ARTICLE

Includes PDF, HTML & Video, when available

Members: $17.00

Non-members: $21.00 ADD TO CART

PROCEEDINGS
PRESENTATION

WATCH
PRESENTATION SAVE TO MY LIBRARY

GET CITATION

KEYWORDS

Biological imaging

Stimulated emission depletion microscopy

Microscopes

Single photon avalanche diodes

Ultrafast phenomena

Fluorescence microscopy

Image enhancement

Show All Keywords

Keywords/Phrases

Search In:

Publication Years