Impact of wavefront distortion and scattering on 2-photon microscopy in mammalian brain tissue

E. Chaigneau, A. J. Wright, S. P. Poland, J. M. Girkin, R. A. Silver

Research output: Contribution to journalArticle

27 Citations (Scopus)

Abstract

Two-photon (2P) microscopy is widely used in neuroscience, but the optical properties of brain tissue are poorly understood. We have investigated the effect of brain tissue on the 2P point spread function (PSF(2P)) by imaging fluorescent beads through living cortical slices. By combining this with measurements of the mean free path of the excitation light, adaptive optics and vector-based modeling that includes phase modulation and scattering, we show that tissue-induced wavefront distortions are the main determinant of enlargement and distortion of the PSF2P at intermediate imaging depths. Furthermore, they generate surrounding lobes that contain more than half of the 2P excitation. These effects reduce the resolution of fine structures and contrast and they, together with scattering, limit 2P excitation. Our results disentangle the contributions of scattering and wavefront distortion in shaping the cortical PSF2P, thereby providing a basis for improved 2P microscopy.
LanguageEnglish
Pages22755-22774
Number of pages20
JournalOptics Express
Volume19
Issue number23
DOIs
Publication statusPublished - 7 Nov 2011

Fingerprint

brain
microscopy
photons
scattering
excitation
neurology
point spread functions
adaptive optics
determinants
phase modulation
beads
lobes
mean free path
fine structure
optical properties

Keywords

  • systems
  • adaptive optics
  • implementation
  • samples
  • specimen-induced aberrations
  • electromagnetic diffraction
  • fluorescence microscopy
  • multiphoton microscopy
  • image field
  • wavefront distortion
  • scattering
  • 2-photon microscopy
  • mammalian brain tissue

Cite this

Chaigneau, E., Wright, A. J., Poland, S. P., Girkin, J. M., & Silver, R. A. (2011). Impact of wavefront distortion and scattering on 2-photon microscopy in mammalian brain tissue. Optics Express, 19(23), 22755-22774. https://doi.org/10.1364/OE.19.022755
Chaigneau, E. ; Wright, A. J. ; Poland, S. P. ; Girkin, J. M. ; Silver, R. A. / Impact of wavefront distortion and scattering on 2-photon microscopy in mammalian brain tissue. In: Optics Express. 2011 ; Vol. 19, No. 23. pp. 22755-22774.
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Chaigneau, E, Wright, AJ, Poland, SP, Girkin, JM & Silver, RA 2011, 'Impact of wavefront distortion and scattering on 2-photon microscopy in mammalian brain tissue' Optics Express, vol. 19, no. 23, pp. 22755-22774. https://doi.org/10.1364/OE.19.022755

Impact of wavefront distortion and scattering on 2-photon microscopy in mammalian brain tissue. / Chaigneau, E.; Wright, A. J.; Poland, S. P.; Girkin, J. M.; Silver, R. A.

In: Optics Express, Vol. 19, No. 23, 07.11.2011, p. 22755-22774.

Research output: Contribution to journalArticle

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