High-fidelity functional and structural whole-brain imaging with Bessel-beam light-sheet microscopy (Conference Presentation)

Marie Caroline Müllenbroich; Ludovico Silvestri; Lapo Turrini; Antonino Paolo Di Giovanna; Tommaso Alterini; Ali Gheisari; Pietro Ricci; Leonardo Sacconi; Francesco Vanzi; Francesco S. Pavone

doi:10.1117/12.2251471

19 April 2017 High-fidelity functional and structural whole-brain imaging with Bessel-beam light-sheet microscopy (Conference Presentation)

Marie Caroline Müllenbroich, Ludovico Silvestri, Lapo Turrini, Antonino Paolo Di Giovanna, Tommaso Alterini, Ali Gheisari, Pietro Ricci, Leonardo Sacconi, Francesco Vanzi, Francesco S. Pavone

Author Affiliations +

Proceedings Volume 10051, Neural Imaging and Sensing; 100510F (2017) https://doi.org/10.1117/12.2251471
Event: SPIE BiOS, 2017, San Francisco, California, United States

Abstract

Light-sheet microscopy (LSM) has proven a useful tool in neuroscience and is particularly well suited to image the entire brain with high frame rates at single cell resolution. On the one hand, LSM is employed in combination with tissue clearing methods like CLARITY which allows for the reconstruction of neuronal or vascular anatomy over cm-sized samples. On the other hand, LSM has been paired with intrinsically transparent samples for real-time recording of neuronal activity with single cell resolution across the entire brain, using calcium indicators like GCaMP6. Despite its intrinsic advantages in terms of high imaging speed and reduced photobleaching, LSM is very sensitive to residual opaque objects present in the sample, which cause dark horizontal stripes in the collected images. In the best case, these artefacts obscure the features of interest in structural imaging; in the worst case, dynamic shadowing introduced by red blood cells significantly alters the fluorescence signal variations related to neuronal activity. We show how the use of Bessel beams in LSM can dramatically reduce such artefacts even in conventional one-sided illumination schemes, thanks to their “self-healing” properties. On the functional side, Bessel-beam LSM allows recording neuronal activity traces without any disturbing flickering caused by the movement of red blood cells. On the structural side, our proposed method is capable of obtaining anatomical information across the entire volume of whole mouse brains allowing tracing blood vessels and neuronal projections also in poorly cleared specimens.

Conference Presentation

Citation Download Citation

Marie Caroline Müllenbroich, Ludovico Silvestri, Lapo Turrini, Antonino Paolo Di Giovanna, Tommaso Alterini, Ali Gheisari, Pietro Ricci, Leonardo Sacconi, Francesco Vanzi, and Francesco S. Pavone "High-fidelity functional and structural whole-brain imaging with Bessel-beam light-sheet microscopy (Conference Presentation)", Proc. SPIE 10051, Neural Imaging and Sensing, 100510F (19 April 2017); https://doi.org/10.1117/12.2251471

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