Arbitrary-scan imaging for two-photon microscopy

Edward Botcherby; Christopher Smith; Martin Booth; Rimas Juskaitis; Tony Wilson

doi:10.1117/12.842134

26 February 2010 Arbitrary-scan imaging for two-photon microscopy

Edward Botcherby, Christopher Smith, Martin Booth, Rimas Juskaitis, Tony Wilson

Proceedings Volume 7569, Multiphoton Microscopy in the Biomedical Sciences X; 756917 (2010) https://doi.org/10.1117/12.842134
Event: SPIE BiOS, 2010, San Francisco, California, United States

Abstract

In this paper, we present details of a scanning two-photon fluorescence microscope we have built with a nearisotropic scan rate. This means that the focal spot can be scanned at high speed along any direction in the specimen, without introducing systematic aberrations. We present experimental point spread function measurements for this system using an Olympus 0.8 NA 40X water dipping objective lens that demonstrates an axial range of operation greater than 200 μm. We give details of a novel actuator device used to displace the focusing element and demonstrate axial scan responses up to 3.5 kHz. Finally, we present a bioscience application of this system to image dendritic processes that follow non-linear paths in three-dimensional space. The focal spot was scanned along one such process at 400 Hz with an axial range of more than 90 μm.

Citation Download Citation

Edward Botcherby, Christopher Smith, Martin Booth, Rimas Juskaitis, and Tony Wilson "Arbitrary-scan imaging for two-photon microscopy", Proc. SPIE 7569, Multiphoton Microscopy in the Biomedical Sciences X, 756917 (26 February 2010); https://doi.org/10.1117/12.842134

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