Special Section on Advanced Biomedical Imaging and Sensing

Multiphoton imaging microscopy at deeper layers with adaptive optics control of spherical aberration

[+] Author Affiliations
Juan M. Bueno, Martin Skorsetz, Raquel Palacios, Emilio J. Gualda, Pablo Artal

Universidad de Murcia, Instituto Universitario de Investigación en Óptica y Nanofísica, Laboratorio de Óptica, Campus de Espianrdo (Ed. 34), 30100 Murcia, Spain

J. Biomed. Opt. 19(1), 011007 (Jul 17, 2013). doi:10.1117/1.JBO.19.1.011007
History: Received March 20, 2013; Revised May 27, 2013; Accepted June 11, 2013
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Abstract.  Despite the inherent confocality and optical sectioning capabilities of multiphoton microscopy, three-dimensional (3-D) imaging of thick samples is limited by the specimen-induced aberrations. The combination of immersion objectives and sensorless adaptive optics (AO) techniques has been suggested to overcome this difficulty. However, a complex plane-by-plane correction of aberrations is required, and its performance depends on a set of image-based merit functions. We propose here an alternative approach to increase penetration depth in 3-D multiphoton microscopy imaging. It is based on the manipulation of the spherical aberration (SA) of the incident beam with an AO device while performing fast tomographic multiphoton imaging. When inducing SA, the image quality at best focus is reduced; however, better quality images are obtained from deeper planes within the sample. This is a compromise that enables registration of improved 3-D multiphoton images using nonimmersion objectives. Examples on ocular tissues and nonbiological samples providing different types of nonlinear signal are presented. The implementation of this technique in a future clinical instrument might provide a better visualization of corneal structures in living eyes.

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© 2014 Society of Photo-Optical Instrumentation Engineers

Citation

Juan M. Bueno ; Martin Skorsetz ; Raquel Palacios ; Emilio J. Gualda and Pablo Artal
"Multiphoton imaging microscopy at deeper layers with adaptive optics control of spherical aberration", J. Biomed. Opt. 19(1), 011007 (Jul 17, 2013). ; http://dx.doi.org/10.1117/1.JBO.19.1.011007


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