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A promising new wavelength region for three-photon fluorescence microscopy of live cells

Norris, Greg and Amor, Rumelo and Dempster, John and Amos, William B and McConnell, Gail (2012) A promising new wavelength region for three-photon fluorescence microscopy of live cells. Journal of Microscopy, 246 (3). pp. 266-273.

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Abstract

We report three-photon laser scanning microscopy (3PLSM) using a bi-directional pumped optical parametric oscillator (OPO) with signal wavelength output at λ= 1500 nm. This novel laser was used to overcome the high optical loss in the infrared spectral region observed in laser scanning microscopes and objective lenses that renders them otherwise difficult to use for imaging. To test our system, we performed 3PLSM auto-fluorescence imaging of live plant cells at λ= 1500 nm, specifically Spirogyra, and compared performance with two-photon excitation (2PLSM) imaging using a femtosecond pulsed Ti:Sapphire laser at λ= 780 nm. Analysis of cell viability based on cytoplasmic organelle streaming and structural changes of cells revealed that at similar peak powers, 2PLSM caused gross cell damage after 5 min but 3PLSM showed little or no interference with cell function after 15 min. The λ= 1500 nm OPO is thus shown to be a practical laser source for live cell imaging.

Item type: Article
ID code: 40251
Notes: © 2012 The Authors Journal of Microscopy © 2012 Wadsworth Center, New York State Department of Health.
Keywords: multi photon, live cell, cytoplasmic streaming, three-photon fluorescence microscopy , Pharmacy and materia medica, Pathology and Forensic Medicine, Histology
Subjects: Medicine > Pharmacy and materia medica
Department: Faculty of Science > Strathclyde Institute of Pharmacy and Biomedical Sciences
Related URLs:
Depositing user: Pure Administrator
Date Deposited: 29 Jun 2012 10:49
Last modified: 27 Mar 2014 10:13
URI: http://strathprints.strath.ac.uk/id/eprint/40251

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