Adaptive optics applied to coherent anti-Stokes Raman scattering microscopy

J.M. Girkin, S.P. Poland, A.J. Wright, C. Freudiger, C.L. Evans, X.S. Xie, A. Periasamy (Editor), P.T.C. So (Editor)

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

We report on the use of adaptive optics in coherent anti-Stokes Raman scattering microscopy (CARS) to improve the image brightness and quality at increased optical penetration depths in biological material. The principle of the technique is to shape the incoming wavefront in such a way that it counteracts the aberrations introduced by imperfect optics and the varying refractive index of the sample. In recent years adaptive optics have been implemented in multiphoton and confocal microscopy. CARS microscopy is proving to be a powerful tool for non-invasive and label-free biomedical imaging with vibrational contrast. As the contrast mechanism is based on a 3rd order non-linear optical process, it is highly susceptible to aberrations, thus CARS signals are commonly lost beyond the depth of∼100 μm in tissue. We demonstrate the combination of adaptive optics and CARS microscopy for deep-tissue imaging using a deformable membrane mirror. A random search optimization algorithm using the CARS intensity as the figure of merit determined the correct mirror-shape in order to correct for the aberrations. We highlight two different methods of implementation, using a look up table technique and by performing the optimizing in situ. We demonstrate a significant increase in brightness and image quality in an agarose/polystyrene-bead sample and white chicken muscle, pushing the penetration depth beyond 200 μm.
Original languageEnglish
Title of host publicationProceedings of SPIE: Multiphoton Microscopy in the Biomedical Sciences VIII
Number of pages436
Volume6860
DOIs
Publication statusPublished - 15 Feb 2008

Fingerprint

adaptive optics
Raman spectra
microscopy
aberration
brightness
penetration
mirrors
chickens
pushing
muscles
figure of merit
beads
polystyrene
optics
refractivity
membranes
optimization

Keywords

  • nonlinear microscopy
  • multi-photon processes
  • biological tissues
  • biological materials
  • penetration depth
  • polystyrene
  • image quality
  • figure of merit
  • refractive index
  • biomedical imaging
  • confocal microscopy
  • optimization method
  • adaptive optics
  • nonlinear optics
  • deformable mirror
  • optical aberration
  • wave front
  • CARS
  • Raman scattering

Cite this

Girkin, J. M., Poland, S. P., Wright, A. J., Freudiger, C., Evans, C. L., Xie, X. S., ... So, P. T. C. (Ed.) (2008). Adaptive optics applied to coherent anti-Stokes Raman scattering microscopy. In Proceedings of SPIE: Multiphoton Microscopy in the Biomedical Sciences VIII (Vol. 6860) https://doi.org/10.1117/12.764445
Girkin, J.M. ; Poland, S.P. ; Wright, A.J. ; Freudiger, C. ; Evans, C.L. ; Xie, X.S. ; Periasamy, A. (Editor) ; So, P.T.C. (Editor). / Adaptive optics applied to coherent anti-Stokes Raman scattering microscopy. Proceedings of SPIE: Multiphoton Microscopy in the Biomedical Sciences VIII. Vol. 6860 2008.
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author = "J.M. Girkin and S.P. Poland and A.J. Wright and C. Freudiger and C.L. Evans and X.S. Xie and A. Periasamy and P.T.C. So",
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Girkin, JM, Poland, SP, Wright, AJ, Freudiger, C, Evans, CL, Xie, XS, Periasamy, A (ed.) & So, PTC (ed.) 2008, Adaptive optics applied to coherent anti-Stokes Raman scattering microscopy. in Proceedings of SPIE: Multiphoton Microscopy in the Biomedical Sciences VIII. vol. 6860. https://doi.org/10.1117/12.764445

Adaptive optics applied to coherent anti-Stokes Raman scattering microscopy. / Girkin, J.M.; Poland, S.P.; Wright, A.J.; Freudiger, C.; Evans, C.L.; Xie, X.S.; Periasamy, A. (Editor); So, P.T.C. (Editor).

Proceedings of SPIE: Multiphoton Microscopy in the Biomedical Sciences VIII. Vol. 6860 2008.

Research output: Chapter in Book/Report/Conference proceedingChapter

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AU - Wright, A.J.

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AU - Xie, X.S.

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N2 - We report on the use of adaptive optics in coherent anti-Stokes Raman scattering microscopy (CARS) to improve the image brightness and quality at increased optical penetration depths in biological material. The principle of the technique is to shape the incoming wavefront in such a way that it counteracts the aberrations introduced by imperfect optics and the varying refractive index of the sample. In recent years adaptive optics have been implemented in multiphoton and confocal microscopy. CARS microscopy is proving to be a powerful tool for non-invasive and label-free biomedical imaging with vibrational contrast. As the contrast mechanism is based on a 3rd order non-linear optical process, it is highly susceptible to aberrations, thus CARS signals are commonly lost beyond the depth of∼100 μm in tissue. We demonstrate the combination of adaptive optics and CARS microscopy for deep-tissue imaging using a deformable membrane mirror. A random search optimization algorithm using the CARS intensity as the figure of merit determined the correct mirror-shape in order to correct for the aberrations. We highlight two different methods of implementation, using a look up table technique and by performing the optimizing in situ. We demonstrate a significant increase in brightness and image quality in an agarose/polystyrene-bead sample and white chicken muscle, pushing the penetration depth beyond 200 μm.

AB - We report on the use of adaptive optics in coherent anti-Stokes Raman scattering microscopy (CARS) to improve the image brightness and quality at increased optical penetration depths in biological material. The principle of the technique is to shape the incoming wavefront in such a way that it counteracts the aberrations introduced by imperfect optics and the varying refractive index of the sample. In recent years adaptive optics have been implemented in multiphoton and confocal microscopy. CARS microscopy is proving to be a powerful tool for non-invasive and label-free biomedical imaging with vibrational contrast. As the contrast mechanism is based on a 3rd order non-linear optical process, it is highly susceptible to aberrations, thus CARS signals are commonly lost beyond the depth of∼100 μm in tissue. We demonstrate the combination of adaptive optics and CARS microscopy for deep-tissue imaging using a deformable membrane mirror. A random search optimization algorithm using the CARS intensity as the figure of merit determined the correct mirror-shape in order to correct for the aberrations. We highlight two different methods of implementation, using a look up table technique and by performing the optimizing in situ. We demonstrate a significant increase in brightness and image quality in an agarose/polystyrene-bead sample and white chicken muscle, pushing the penetration depth beyond 200 μm.

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KW - wave front

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KW - Raman scattering

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M3 - Chapter

SN - 9780819470355

VL - 6860

BT - Proceedings of SPIE: Multiphoton Microscopy in the Biomedical Sciences VIII

ER -

Girkin JM, Poland SP, Wright AJ, Freudiger C, Evans CL, Xie XS et al. Adaptive optics applied to coherent anti-Stokes Raman scattering microscopy. In Proceedings of SPIE: Multiphoton Microscopy in the Biomedical Sciences VIII. Vol. 6860. 2008 https://doi.org/10.1117/12.764445