Sensorless adaptive optics optical coherence tomography for two photon excited fluorescence mouse retinal imaging

Daniel J. Wahl, Myeong Jin Ju, Yifan Jian, Marinko V. Sarunic

Research output: Chapter in Book/Report/Conference proceedingConference contribution

Abstract

Two-Photon Excited Fluorescence (TPEF) is a common modality for volumetric imaging within a biological sample. The molecule-specific contrast of TPEF imaging of the retina enables novel in vivo studies of disease and retina physiology. Furthermore, retinal studies can be aided by imaging the relevant cells, which can be fluorescently labelled with fluorophores such as GFP. The multiphoton absorption suppresses the out-of-focus background signal and improves with the axial sectioning without a confocal aperture in the optical detection path. Imaging the retina with near infrared (NIR) light is ideal since the retina contain visual pigments that are sensitive to visible wavelengths and NIR light has less scattering within biological tissue than shorter wavelengths. However, high pulse energy is required to generate the TPEF, yet minimizing the incident exposure energy is required for non-invasive imaging. TPEF signal intensity increases quartically with the spot size, which leads to high sensitivity to aberrations that distribute the energy deposition of the focused light1. For retinal imaging, decreasing the spot size requires the imaging beam to fill a larger area of the eye, which corresponds to an increase in the aberration amplitudes and often results in low TPEF. Recent research has demonstrated improvements to the TPEF signal for retinal imaging using AO for aberrations correction2,3,4. Theoretically, the numerical aperture through the pupil of the mouse eye permits sub-micrometer imaging of the retina. However, optical aberrations introduced by the tear film, cornea and intraocular lens reduce the actual resolution. In order to approach diffraction limited imaging, these aberrations can be corrected with Adaptive Optics (AO) using a wavefront corrector such as Deformable Mirror (DM). The traditional approach to AO is to use a Wavefront Sensor (WFS) to measure the ocular aberrations directly. Performing accurate wavefront measurements for WFS AO imaging in a small animal eye requires a high system complexity due to the short length of the eye creating an optically thick sample with multiple scattering surfaces. Alternatively, Sensorless AO (SAO) has the potential to overcome the limitations of the SH-WFS. SAO does not require direct measurement of the optical wavefront but instead uses an image-based approach. SAO methods have the ability to provide depth resolved aberration correction by performing the aberration correction at different layers within the retina. Our imaging system provides volumetric TPEF imaging in the retina using SAO-OCT for depth-specific aberration correction, using same light source to generate the OCT and TPEF. Here, we present our progress since our previous report4 with improvements to the light delivery, aberration correction, and TPEF detection.

Original languageEnglish (US)
Title of host publicationOptical Coherence Imaging Techniques and Imaging in Scattering Media III
EditorsMaciej Wojtkowski, Stephen A. Boppart, Wang-Yuhl Oh
PublisherSPIE
ISBN (Electronic)9781510628496
DOIs
StatePublished - Jan 1 2019
EventOptical Coherence Imaging Techniques and Imaging in Scattering Media III 2019 - Munich, Germany
Duration: Jun 25 2019Jun 27 2019

Publication series

NameProgress in Biomedical Optics and Imaging - Proceedings of SPIE
Volume11078
ISSN (Print)1605-7422

Conference

ConferenceOptical Coherence Imaging Techniques and Imaging in Scattering Media III 2019
CountryGermany
CityMunich
Period6/25/196/27/19

Fingerprint

Adaptive optics
Optical tomography
Optical Coherence Tomography
Photons
adaptive optics
mice
aberration
Aberrations
retina
tomography
Fluorescence
Retina
Imaging techniques
fluorescence
photons
Wavefronts
Light
Optical Imaging
sensors
visual pigments

ASJC Scopus subject areas

  • Electronic, Optical and Magnetic Materials
  • Biomaterials
  • Atomic and Molecular Physics, and Optics
  • Radiology Nuclear Medicine and imaging

Cite this

Wahl, D. J., Ju, M. J., Jian, Y., & Sarunic, M. V. (2019). Sensorless adaptive optics optical coherence tomography for two photon excited fluorescence mouse retinal imaging. In M. Wojtkowski, S. A. Boppart, & W-Y. Oh (Eds.), Optical Coherence Imaging Techniques and Imaging in Scattering Media III [110781L] (Progress in Biomedical Optics and Imaging - Proceedings of SPIE; Vol. 11078). SPIE. https://doi.org/10.1117/12.2527203

Sensorless adaptive optics optical coherence tomography for two photon excited fluorescence mouse retinal imaging. / Wahl, Daniel J.; Ju, Myeong Jin; Jian, Yifan; Sarunic, Marinko V.

Optical Coherence Imaging Techniques and Imaging in Scattering Media III. ed. / Maciej Wojtkowski; Stephen A. Boppart; Wang-Yuhl Oh. SPIE, 2019. 110781L (Progress in Biomedical Optics and Imaging - Proceedings of SPIE; Vol. 11078).

Research output: Chapter in Book/Report/Conference proceedingConference contribution

Wahl, DJ, Ju, MJ, Jian, Y & Sarunic, MV 2019, Sensorless adaptive optics optical coherence tomography for two photon excited fluorescence mouse retinal imaging. in M Wojtkowski, SA Boppart & W-Y Oh (eds), Optical Coherence Imaging Techniques and Imaging in Scattering Media III., 110781L, Progress in Biomedical Optics and Imaging - Proceedings of SPIE, vol. 11078, SPIE, Optical Coherence Imaging Techniques and Imaging in Scattering Media III 2019, Munich, Germany, 6/25/19. https://doi.org/10.1117/12.2527203
Wahl DJ, Ju MJ, Jian Y, Sarunic MV. Sensorless adaptive optics optical coherence tomography for two photon excited fluorescence mouse retinal imaging. In Wojtkowski M, Boppart SA, Oh W-Y, editors, Optical Coherence Imaging Techniques and Imaging in Scattering Media III. SPIE. 2019. 110781L. (Progress in Biomedical Optics and Imaging - Proceedings of SPIE). https://doi.org/10.1117/12.2527203
Wahl, Daniel J. ; Ju, Myeong Jin ; Jian, Yifan ; Sarunic, Marinko V. / Sensorless adaptive optics optical coherence tomography for two photon excited fluorescence mouse retinal imaging. Optical Coherence Imaging Techniques and Imaging in Scattering Media III. editor / Maciej Wojtkowski ; Stephen A. Boppart ; Wang-Yuhl Oh. SPIE, 2019. (Progress in Biomedical Optics and Imaging - Proceedings of SPIE).
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