Two-photon microscopy with dynamic focusing objective using a liquid lens

A variable focus microscope without moving parts for axial focusing within the sample in two-photon microscopy can provide a faster, more robust, and cost-effective solution to 3D imaging at high resolution. A custom dynamic focusing objective was recently developed that incorporates a liquid lens within the optical design of a custom microscope. In this paper, we investigate the use of a liquid lens embedded in an aberration-corrected custom microscope for z-dimension scanning in 3D resolved two-photon microscopy. Results show that we can achieve imaging speed of 50 frames per sec for an arbitrary plane in 3D space, a leap forward in scan speed for two-photon microscopy.

[1]  Supraja Murali,et al.  Three-dimensional adaptive microscopy using embedded liquid lens. , 2009, Optics letters.

[2]  I. Parker,et al.  Construction of a confocal microscope for real-time x-y and x-z imaging. , 1999, Cell calcium.

[3]  I. Alex Vitkin,et al.  Dynamic focus control in high-speed optical coherence tomography based on a microelectromechanical mirror , 2004 .

[4]  Masatoshi Ishikawa,et al.  Variable-focus lens with 1-kHz bandwidth. , 2004, Optics express.

[5]  P. So,et al.  Two-Photon deep tissue ex vivo imaging of mouse dermal and subcutaneous structures. , 1998, Optics express.

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

[7]  K. Svoboda,et al.  Long-term in vivo imaging of experience-dependent synaptic plasticity in adult cortex , 2002, Nature.

[8]  David S. Greenberg,et al.  Imaging input and output of neocortical networks in vivo. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[9]  W. Denk,et al.  Deep tissue two-photon microscopy , 2005, Nature Methods.

[10]  N. Kasthuri,et al.  Long-term dendritic spine stability in the adult cortex , 2002, Nature.

[11]  P. Meemon,et al.  Gabor domain optical coherence microscopy , 2008, BiOS.

[12]  K. Fujita.  [Two-photon laser scanning fluorescence microscopy]. , 2007, Tanpakushitsu kakusan koso. Protein, nucleic acid, enzyme.

[13]  F. Helmchen,et al.  Imaging cellular network dynamics in three dimensions using fast 3D laser scanning , 2007, Nature Methods.

[14]  Panomsak Meemon,et al.  Assessment of a liquid lens enabled in vivo optical coherence microscope. , 2010, Applied optics.