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Proceedings Paper

Practical optical quality assessment and correction of a nonlinear microscope
Author(s): Rodrigo Aviles-Espinosa; Jordi Andilla; Rafael Porcar-Guezenec; Omar Olarte; Susana I.C. O. Santos; Xavier Levecq; David Artigas; Pablo Loza-Alvarez
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Paper Abstract

Nonlinear microscopy (NLM) has covered the requirement for higher contrast and resolution compared with other microscopy techniques, however, the optical quality of this imaging apparatus and the sample structure can compromise its capabilities. Here, we show that the imaging capabilities of a NLM can be affected by the aberrations produced by the setup optical elements alignment, the materials from which they are fabricated and more importantly by the sample. To overcome this, a Shack-Hartman Wavefront sensing scheme has been implemented for characterizing: a) the whole NLM setup and, b) the sample induced aberrations. The first part includes all the aberrations introduced by the optical elements, starting from the laser and until the microscope objective. Having these information, aberrations can be compensated in a closed-loop configuration resulting in the system calibration. Then the remaining aberrations (microscope objective and sample) are recorded. This is done employing the sample nonlinear fluorescence signal collected at one point (keeping the excitation beam static) in the imaging plane. Given that this emission is an incoherent process, it can be considered as a point source. Therefore its wavefront will contain the sample and the objective aberrations. Using the wavefront sensor the information is recorded and passed to the deformable mirror which will compensate the aberrations in a "single shot" (open-loop configuration). This compared with other adaptive optics strategies (i.e. iterative algorithms) results in a reduced sample exposure, and greatly decreases sample damage. Importantly the application of both corrections (system and sample) enables a significant signal intensity and contrast improvement.

Paper Details

Date Published: 24 February 2010
PDF: 8 pages
Proc. SPIE 7570, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XVII, 75700W (24 February 2010); doi: 10.1117/12.840978
Show Author Affiliations
Rodrigo Aviles-Espinosa, ICFO-The Institute of Photonic Sciences (Spain)
Jordi Andilla, Imagine Optic (France)
Rafael Porcar-Guezenec, Imagine Optic (France)
ICFO-The Institute of Photonic Sciences (Spain)
Omar Olarte, ICFO-The Institute of Photonic Sciences (Spain)
Susana I.C. O. Santos, ICFO-The Institute of Photonic Sciences (Spain)
Xavier Levecq, Imagine Optic (France)
David Artigas, Univ. Politècnica de Catalunya (Spain)
ICFO-The Institute of Photonic Sciences (Spain)
Pablo Loza-Alvarez, ICFO-The Institute of Photonic Sciences (Spain)

Published in SPIE Proceedings Vol. 7570:
Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XVII
Jose-Angel Conchello; Carol J. Cogswell; Tony Wilson; Thomas G. Brown, Editor(s)

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