Continuous focal translation enhances rate of point-scan volumetric microscopy.


Journal Article

Two-Photon Laser-Scanning Microscopy is a powerful tool for exploring biological structure and function due to its ability to optically section through a sample with a tight focus. While it is possible to obtain 3D image stacks by moving a stage, this per-frame imaging process is time consuming. Here, we present a method for an easy-to-implement and inexpensive modification of an existing two-photon microscope to rapidly image in 3D using an electrically tunable lens to create a tessellating scan pattern which repeats with the volume rate. Using appropriate interpolating algorithms, the volumetric imaging rate can be increased by a factor up to four-fold. This capability provides the expansion of the two-photon microscope into the third dimension for faster volumetric imaging capable of visualizing dynamics on timescales not achievable by traditional stage-stack methods.

Full Text

Duke Authors

Cited Authors

  • Johnson, C; Exell, J; Kuo, J; Welsher, K

Published Date

  • December 2019

Published In

Volume / Issue

  • 27 / 25

Start / End Page

  • 36241 - 36258

PubMed ID

  • 31873407

Pubmed Central ID

  • 31873407

Electronic International Standard Serial Number (EISSN)

  • 1094-4087

International Standard Serial Number (ISSN)

  • 1094-4087

Digital Object Identifier (DOI)

  • 10.1364/oe.27.036241


  • eng