The effects of refractive index heterogeneity within kidney tissue on multiphoton fluorescence excitation microscopy

P. A. Young, S. G. Clendenon, J. M. Byars, K. W. Dunn

Research output: Contribution to journalArticle

11 Scopus citations

Abstract

Although multiphoton fluorescence excitation microscopy has improved the depth at which useful fluorescence images can be collected in biological tissues, the reach of multiphoton fluorescence excitation microscopy is nonetheless limited by tissue scattering and spherical aberration. Scattering can be reduced in fixed samples by mounting in a medium whose refractive index closely matches that of the fixed material. Using optical 'clearing', the effects of refractive index heterogeneity on signal attenuation with depth are investigated. Quantitative measurements show that by mounting kidney tissue in a high refractive index medium, less than 50% of signal attenuates in 100 μm of depth.

Original languageEnglish (US)
Pages (from-to)148-156
Number of pages9
JournalJournal of Microscopy
Volume242
Issue number2
DOIs
StatePublished - May 1 2011

Keywords

  • Multiphoton fluorescence excitation microscopy
  • Refractive index heterogeneity
  • Refractive index mismatch
  • Scattering
  • Spherical aberration
  • Two-photon microscopy

ASJC Scopus subject areas

  • Histology
  • Pathology and Forensic Medicine

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