Electron microscopic measurement of the size of the optical focus in laser scanning microscopy

Research output: Contribution to journalArticle

Abstract

We describe a method for measuring the lateral focal spot size of a multiphoton laser scanning microscope (LSM) with unprecedented accuracy. A specimen consisting of an aluminum film deposited on a glass coverslip was brought into focus in a LSM and the laser intensity was then increased enough to perform nanoablation of the metal film. This process leaves a permanent trace of the raster path usually taken by the beam during the acquisition of an optical image. A scanning electron microscope (SEM) was then used to determine the nanoablated line width to high accuracy, from which the lateral spot size and hence resolution of the LSM can be determined. To demonstrate our method, we performed analysis of a multiphoton LSM at various infrared wavelengths, and we report measurements of optical lateral spot size with an accuracy of 20 nm, limited only by the resolution of the SEM.
LanguageEnglish
Pages596-602
Number of pages7
JournalMicroscopy and Microanalysis
Volume18
Issue number3
DOIs
Publication statusPublished - Jun 2012

Fingerprint

Microscopic examination
microscopy
Scanning
scanning
Electrons
Lasers
Microscopes
microscopes
lasers
electrons
Electron microscopes
electron microscopes
metal films
Linewidth
leaves
acquisition
Infrared radiation
aluminum
Aluminum
Glass

Keywords

  • electron microscopic measurement
  • optical focus
  • lasers
  • laser scanning
  • laser scanning microscopy

Cite this

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abstract = "We describe a method for measuring the lateral focal spot size of a multiphoton laser scanning microscope (LSM) with unprecedented accuracy. A specimen consisting of an aluminum film deposited on a glass coverslip was brought into focus in a LSM and the laser intensity was then increased enough to perform nanoablation of the metal film. This process leaves a permanent trace of the raster path usually taken by the beam during the acquisition of an optical image. A scanning electron microscope (SEM) was then used to determine the nanoablated line width to high accuracy, from which the lateral spot size and hence resolution of the LSM can be determined. To demonstrate our method, we performed analysis of a multiphoton LSM at various infrared wavelengths, and we report measurements of optical lateral spot size with an accuracy of 20 nm, limited only by the resolution of the SEM.",
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Electron microscopic measurement of the size of the optical focus in laser scanning microscopy. / McConnell, Gail.

In: Microscopy and Microanalysis, Vol. 18, No. 3, 06.2012, p. 596-602.

Research output: Contribution to journalArticle

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