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Optics Express

Optics Express

  • Editor: Andrew M. Weiner
  • Vol. 22, Iss. 16 — Aug. 11, 2014
  • pp: 19293–19301

Simultaneous multiplane confocal microscopy using acoustic tunable lenses

Martí Duocastella, Giuseppe Vicidomini, and Alberto Diaspro  »View Author Affiliations

Optics Express, Vol. 22, Issue 16, pp. 19293-19301 (2014)

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Maximizing the amount of spatiotemporal information retrieved in confocal laser scanning microscopy is crucial to understand fundamental three-dimensional (3D) dynamic processes in life sciences. However, current 3D confocal microscopy is based on an inherently slow stepwise process that consists of acquiring multiple 2D sections at different focal planes by mechanical or optical z-focus translation. Here, we show that by using an acoustically-driven optofluidic lens integrated in a commercial confocal system we can capture an entire 3D image in a single step. Our method is based on continuous axial scanning at speeds as high as 140 kHz combined with fast readout. In this way, one or more focus sweeps are produced on a pixel by pixel basis and the detected photons can be assigned to their corresponding focal plane enabling simultaneous multiplane imaging. We exemplify this method by imaging calibration and biological fluorescence samples. These results open the door to exploring new fundamental processes in science with an unprecedented time resolution.

© 2014 Optical Society of America

OCIS Codes
(170.1790) Medical optics and biotechnology : Confocal microscopy
(180.0180) Microscopy : Microscopy
(110.1080) Imaging systems : Active or adaptive optics

ToC Category:

Original Manuscript: June 23, 2014
Revised Manuscript: July 25, 2014
Manuscript Accepted: July 25, 2014
Published: August 4, 2014

Virtual Issues
Vol. 9, Iss. 10 Virtual Journal for Biomedical Optics

Martí Duocastella, Giuseppe Vicidomini, and Alberto Diaspro, "Simultaneous multiplane confocal microscopy using acoustic tunable lenses," Opt. Express 22, 19293-19301 (2014)

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