Light field multiphoton microscopy with temporal focusing-based volume selective excitation

Feng-Chun Hsu; Yong Da Sie; Chun-Yu Lin; Yvonne Yuling Hu; Shean-Jen Chen

doi:10.1117/12.2621688

24 May 2022 Light field multiphoton microscopy with temporal focusing-based volume selective excitation

Feng-Chun Hsu, Yong Da Sie, Chun-Yu Lin, Yvonne Yuling Hu, Shean-Jen Chen

Proceedings Volume 12136, Unconventional Optical Imaging III; 121360F (2022) https://doi.org/10.1117/12.2621688
Event: SPIE Photonics Europe, 2022, Strasbourg, France

Abstract

Light field fluorescence microscopy (LFM) can provide three-dimensional (3D) images in one snapshot, but essentially lighting up the entirety of the sample, even though only a part of the sample is meaningfully captured in the reconstruction. Thus, entire illumination introduces extraneous background noise, degrading the contrast and accuracy of the final reconstructed images. In this paper, temporal focusing-based multiphoton illumination (TFMI) has the advantage of widefield multiphoton excitation with volume selective excitation. We implement the TFMI to LFM, illuminating only the volume of interest, thus significantly reducing the background. Furthermore, offering higher penetration depth in scattering tissue via multiphoton. In addition, the volume range can be varied by modulating the size of the Fourier-plane aperture of objective lens. 100 nm fluorescence beads are used to examine the lateral and axial resolution after phase space deconvolution from light field image, the experimental results show that the lateral resolution is around 1.2 μm and axial resolution is around 1.6 μm close to the focal plane. Furthermore, the mushroom body of drosophila brain which carried a genetic fluorescent marker GFP (OK-107) are used to demonstrate volumetric bioimaging capability.

Conference Presentation

Citation Download Citation

Feng-Chun Hsu, Yong Da Sie, Chun-Yu Lin, Yvonne Yuling Hu, and Shean-Jen Chen "Light field multiphoton microscopy with temporal focusing-based volume selective excitation", Proc. SPIE 12136, Unconventional Optical Imaging III, 121360F (24 May 2022); https://doi.org/10.1117/12.2621688

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