Fast 3-D temporal focusing microscopy using an electrically tunable lens.

In this paper, we present a 3-D temporal focusing microscope based on an electrically tunable lens (ETL) and a femtosecond regenerative laser amplifier. The focus-tunable lens provides a fast and compact way to perform non-mechanical z-scanning and resolves the blurry image issue compared with GVD-based z-scanning methods. The optical performance of the temporal focusing system, including z-scanning characteristics, the associated the magnification variation, and the lateral and axial resolution, has been studied and characterized using calibrated Rhodamine-6G thin film sample, fluorescent beads, and pollen samples. Lastly, we demonstrate the optical cross-sectioning and z-scanning capability with an in vivo experiment, where Ca(2+) imaging of neurons in GaCamp6 labeled zebrafish was performed.

[1]  Y. Silberberg,et al.  Scanningless depth-resolved microscopy. , 2005, Optics express.

[2]  F. El-orany.  Atomic Wehrl entropy for the Jaynes–Cummings model: explicit form and Bloch sphere radius , 2009 .

[3]  W. Zipfel,et al.  Simultaneous spatial and temporal focusing of femtosecond pulses , 2005, (CLEO). Conference on Lasers and Electro-Optics, 2005..

[4]  S. Hell,et al.  Multifocal multiphoton microscopy. , 1998, Optics letters.

[5]  P. So,et al.  High-speed, two-photon scanning microscope. , 1999, Applied optics.

[6]  M H Ellisman,et al.  Video-rate scanning two-photon excitation fluorescence microscopy and ratio imaging with cameleons. , 1999, Biophysical journal.

[7]  Benjamin Schmid,et al.  Rapid 3D light-sheet microscopy with a tunable lens. , 2013, Optics express.

[8]  W. Denk,et al.  Two-photon laser scanning fluorescence microscopy. , 1990, Science.

[9]  Chris Xu,et al.  High speed multiphoton axial scanning through an optical fiber in a remotely scanned temporal focusing setup , 2010, Biomedical optics express.

[10]  I. Parker,et al.  Construction of a two-photon microscope for video-rate Ca(2+) imaging. , 2001, Cell calcium.

[11]  Benjamin F. Grewe,et al.  Fast two-layer two-photon imaging of neuronal cell populations using an electrically tunable lens , 2011, Biomedical optics express.

[12]  Benjamin F. Grewe,et al.  Cellular Level Brain Imaging in Behaving Mammals: An Engineering Approach , 2015, Neuron.

[13]  S. Shoham,et al.  Remotely scanned multiphoton temporal focusing by axial grism scanning. , 2012, Optics letters.

[14]  M. Durst,et al.  Simultaneous spatial and temporal focusing for axial scanning. , 2006, Optics express.

[15]  S. Zeng,et al.  Analysis of fast axial scanning scheme using temporal focusing with acousto-optic deflectors , 2009 .

[16]  T Nielsen,et al.  High efficiency beam splitter for multifocal multiphoton microscopy , 2001, Journal of microscopy.

[17]  Buist,et al.  Real time two‐photon absorption microscopy using multi point excitation , 1998 .

[18]  J. Squier,et al.  Widefield multiphoton and temporally decorrelated multifocal multiphoton microscopy. , 2000, Optics express.

[19]  S W Hell,et al.  Space‐multiplexed multifocal nonlinear microscopy , 2001, Journal of microscopy.

[20]  O. Paulsen,et al.  Aberration-free three-dimensional multiphoton imaging of neuronal activity at kHz rates , 2012, Proceedings of the National Academy of Sciences.