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Enhanced volumetric imaging in 2-photon microscopy via acoustic lens beam shaping
- Source :
- Journal of biophotonics. 11(2)
- Publication Year :
- 2017
-
Abstract
- Three-dimensional imaging at high-spatiotemporal resolutions and over large penetration depths is key for unmasking the dynamics and structural organization of complex biological systems. However, the need to axially shift the focus, with consequent limitations in imaging speed, and signal degradation at large depths due to scattering effects, makes this task challenging. Here, we present a novel approach in 2-photon excitation microscopy that allows fast volumetric imaging and enhanced signal-to-background (S/B) in thick tissue. Our technique is based on ultrafast beam shaping at each pixel by means of an acoustic optofluidic lens. Shaping the excitation beam with different phase profiles enables both high-speed axial focus shifting, for continuous volumetric imaging, and controlled aberrated imaging, advantageous for out-of-focus background removal. We provide a theoretical description of our approach, and demonstrate volumetric imaging of neuronal cells from a mouse brain slice with enhancements in S/B up to a factor of 10 over a depth of 600 μm.
- Subjects :
- Genetics and Molecular Biology (all)
0301 basic medicine
Acousto-optic lens
Adaptive optics
Optofluidics
Three-dimensional imaging
Two-photon microscopy
Chemistry (all)
Materials Science (all)
Biochemistry, Genetics and Molecular Biology (all)
Engineering (all)
Physics and Astronomy (all)
Materials science
General Physics and Astronomy
Signal-To-Noise Ratio
Biochemistry
01 natural sciences
General Biochemistry, Genetics and Molecular Biology
010309 optics
03 medical and health sciences
Optics
Imaging, Three-Dimensional
Two-photon excitation microscopy
0103 physical sciences
Microscopy
Scattering, Radiation
General Materials Science
Lenses
Pixel
Scattering
business.industry
Phantoms, Imaging
General Engineering
General Chemistry
Acoustics
030104 developmental biology
Microscopy, Fluorescence, Multiphoton
Feasibility Studies
business
Ultrashort pulse
Excitation
Subjects
Details
- ISSN :
- 18640648
- Volume :
- 11
- Issue :
- 2
- Database :
- OpenAIRE
- Journal :
- Journal of biophotonics
- Accession number :
- edsair.doi.dedup.....5fd085980e941e710a2524cfb8842a05