Adaptive optics confocal fluorescence microscopy with direct wavefront sensing for brain tissue imaging

Recently, there has been a growing interest in deep tissue imaging for the study of neurons. Unfortunately, because of the inhomogeneous refractive index of the tissue, the aberrations degrade the resolution and brightness of the final image. In this paper, we describe an adaptive optics confocal fluorescence microscope (AOCFM) which can correct aberrations based on direct wavefront measurements using a point source reference beacon and a Shack-Hartmann Wavefront Sensor (SHWS). Mouse brain tissues with different thicknesses are tested. After correction, both the signal intensity and contrast of the image are improved.

[1]  J. Pawley,et al.  Handbook of Biological Confocal Microscopy , 1990, Springer US.

[2]  Robert K. Tyson.  Principles of Adaptive Optics , 1991 .

[3]  J Beuthan,et al.  The spatial variation of the refractive index in biological cells. , 1996, Physics in medicine and biology.

[4]  G. Mourou,et al.  Smart microscope: an adaptive optics learning system for aberration correction in multiphoton confocal microscopy. , 2000, Optics letters.

[5]  Kenzo Hirose,et al.  Visualization of IP3 Dynamics Reveals a Novel AMPA Receptor-Triggered IP3 Production Pathway Mediated by Voltage-Dependent Ca2+ Influx in Purkinje Cells , 2001, Neuron.

[6]  Botond Roska,et al.  Tuning FlaSh: redesign of the dynamics, voltage range, and color of the genetically encoded optical sensor of membrane potential. , 2002, Biophysical journal.

[7]  S. Quake,et al.  Number, Density, and Surface/Cytoplasmic Distribution of GABA Transporters at Presynaptic Structures of Knock-In Mice Carrying GABA Transporter Subtype 1–Green Fluorescent Protein Fusions , 2002, The Journal of Neuroscience.

[8]  Lisa A Poyneer,et al.  Fast wave-front reconstruction in large adaptive optics systems with use of the Fourier transform. , 2002, Journal of the Optical Society of America. A, Optics, image science, and vision.

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

[10]  J. Girkin,et al.  Practical implementation of adaptive optics in multiphoton microscopy. , 2003, Optics express.

[11]  W. Denk,et al.  Adaptive wavefront correction in two-photon microscopy using coherence-gated wavefront sensing , 2006, Proceedings of the National Academy of Sciences.

[12]  Gerard Rousset,et al.  Comparison of centroid computation algorithms in a Shack–Hartmann sensor , 2006 .

[13]  Abdul Ahad S. Awwal,et al.  Adaptive Optics for Vision Science: Principles, Practices, Design, and Applications , 2006 .

[14]  W. Gan,et al.  Choice of cranial window type for in vivo imaging affects dendritic spine turnover in the cortex , 2007, Nature Neuroscience.

[15]  M. Booth.  Adaptive optics in microscopy. , 2003, Philosophical transactions. Series A, Mathematical, physical, and engineering sciences.

[16]  Tony Wilson,et al.  Image-based adaptive optics for two-photon microscopy. , 2009, Optics letters.

[17]  Daren Dillon,et al.  Wavefront aberration measurements and corrections through thick tissue using fluorescent microsphere reference beacons , 2010, Optics express.

[18]  Eric Betzig,et al.  Adaptive optics via pupil segmentation for high-resolution imaging in biological tissues , 2010, Nature Methods.