Adaptive-optics SLO imaging combined with widefield OCT and SLO enables precise 3D localization of fluorescent cells in the mouse retina

Robert Zawadzki, Pengfei Zhang, Azhar Zam, Eric B. Miller, Mayank Goswami, Xinlei Wang, Ravi Jonnal, Sang Hyuck Lee, Dae Yu Kim, John G. Flannery, John S Werner, Marie E Burns, Edward N Pugh Jr

Research output: Contribution to journalArticle

29 Citations (Scopus)

Abstract

Adaptive optics scanning laser ophthalmoscopy (AO-SLO) has recently been used to achieve exquisite subcellular resolution imaging of the mouse retina. Wavefront sensing-based AO typically restricts the field of view to a few degrees of visual angle. As a consequence the relationship between AO-SLO data and larger scale retinal structures and cellular patterns can be difficult to assess. The retinal vasculature affords a largescale 3D map on which cells and structures can be located during in vivo imaging. Phase-variance OCT (pv-OCT) can efficiently image the vasculature with near-infrared light in a label-free manner, allowing 3D vascular reconstruction with high precision. We combined widefield pv-OCT and SLO imaging with AO-SLO reflection and fluorescence imaging to localize two types of fluorescent cells within the retinal layers: GFP-expressing microglia, the resident macrophages of the retina, and GFP-expressing cone photoreceptor cells. We describe in detail a reflective afocal AO-SLO retinal imaging system designed for high resolution retinal imaging in mice. The optical performance of this instrument is compared to other state-of-the-art AO-based mouse retinal imaging systems. The spatial and temporal resolution of the new AO instrumentation was characterized with angiography of retinal capillaries, including blood-flow velocity analysis. Depth-resolved AO-SLO fluorescent images of microglia and cone photoreceptors are visualized in parallel with 469 nm and 663 nm reflectance images of the microvasculature and other structures. Additional applications of the new instrumentation are discussed.

Original languageEnglish (US)
Pages (from-to)2191-2210
Number of pages20
JournalBiomedical Optics Express
Volume6
Issue number6
DOIs
StatePublished - May 21 2015

Fingerprint

Ophthalmoscopy
retina
adaptive optics
mice
Retina
Lasers
scanning
Retinal Cone Photoreceptor Cells
cells
photoreceptors
lasers
Microglia
cones
macrophages
Blood Flow Velocity
angiography
Optical Imaging
Cellular Structures
blood flow
Microvessels

Keywords

  • Active or adaptive optics
  • Imaging systems
  • Ophthalmic optics and devices
  • Ophthalmology
  • Optical coherence tomography
  • Visual optics, comparative animal models

ASJC Scopus subject areas

  • Atomic and Molecular Physics, and Optics
  • Biotechnology

Cite this

Adaptive-optics SLO imaging combined with widefield OCT and SLO enables precise 3D localization of fluorescent cells in the mouse retina. / Zawadzki, Robert; Zhang, Pengfei; Zam, Azhar; Miller, Eric B.; Goswami, Mayank; Wang, Xinlei; Jonnal, Ravi; Lee, Sang Hyuck; Kim, Dae Yu; Flannery, John G.; Werner, John S; Burns, Marie E; Pugh Jr, Edward N.

In: Biomedical Optics Express, Vol. 6, No. 6, 21.05.2015, p. 2191-2210.

Research output: Contribution to journalArticle

Zawadzki, Robert ; Zhang, Pengfei ; Zam, Azhar ; Miller, Eric B. ; Goswami, Mayank ; Wang, Xinlei ; Jonnal, Ravi ; Lee, Sang Hyuck ; Kim, Dae Yu ; Flannery, John G. ; Werner, John S ; Burns, Marie E ; Pugh Jr, Edward N. / Adaptive-optics SLO imaging combined with widefield OCT and SLO enables precise 3D localization of fluorescent cells in the mouse retina. In: Biomedical Optics Express. 2015 ; Vol. 6, No. 6. pp. 2191-2210.
@article{294d7253fea94a8c886a6fc2df514200,
title = "Adaptive-optics SLO imaging combined with widefield OCT and SLO enables precise 3D localization of fluorescent cells in the mouse retina",
abstract = "Adaptive optics scanning laser ophthalmoscopy (AO-SLO) has recently been used to achieve exquisite subcellular resolution imaging of the mouse retina. Wavefront sensing-based AO typically restricts the field of view to a few degrees of visual angle. As a consequence the relationship between AO-SLO data and larger scale retinal structures and cellular patterns can be difficult to assess. The retinal vasculature affords a largescale 3D map on which cells and structures can be located during in vivo imaging. Phase-variance OCT (pv-OCT) can efficiently image the vasculature with near-infrared light in a label-free manner, allowing 3D vascular reconstruction with high precision. We combined widefield pv-OCT and SLO imaging with AO-SLO reflection and fluorescence imaging to localize two types of fluorescent cells within the retinal layers: GFP-expressing microglia, the resident macrophages of the retina, and GFP-expressing cone photoreceptor cells. We describe in detail a reflective afocal AO-SLO retinal imaging system designed for high resolution retinal imaging in mice. The optical performance of this instrument is compared to other state-of-the-art AO-based mouse retinal imaging systems. The spatial and temporal resolution of the new AO instrumentation was characterized with angiography of retinal capillaries, including blood-flow velocity analysis. Depth-resolved AO-SLO fluorescent images of microglia and cone photoreceptors are visualized in parallel with 469 nm and 663 nm reflectance images of the microvasculature and other structures. Additional applications of the new instrumentation are discussed.",
keywords = "Active or adaptive optics, Imaging systems, Ophthalmic optics and devices, Ophthalmology, Optical coherence tomography, Visual optics, comparative animal models",
author = "Robert Zawadzki and Pengfei Zhang and Azhar Zam and Miller, {Eric B.} and Mayank Goswami and Xinlei Wang and Ravi Jonnal and Lee, {Sang Hyuck} and Kim, {Dae Yu} and Flannery, {John G.} and Werner, {John S} and Burns, {Marie E} and {Pugh Jr}, {Edward N}",
year = "2015",
month = "5",
day = "21",
doi = "10.1364/BOE.6.002191",
language = "English (US)",
volume = "6",
pages = "2191--2210",
journal = "Biomedical Optics Express",
issn = "2156-7085",
publisher = "The Optical Society",
number = "6",

}

TY - JOUR

T1 - Adaptive-optics SLO imaging combined with widefield OCT and SLO enables precise 3D localization of fluorescent cells in the mouse retina

AU - Zawadzki, Robert

AU - Zhang, Pengfei

AU - Zam, Azhar

AU - Miller, Eric B.

AU - Goswami, Mayank

AU - Wang, Xinlei

AU - Jonnal, Ravi

AU - Lee, Sang Hyuck

AU - Kim, Dae Yu

AU - Flannery, John G.

AU - Werner, John S

AU - Burns, Marie E

AU - Pugh Jr, Edward N

PY - 2015/5/21

Y1 - 2015/5/21

N2 - Adaptive optics scanning laser ophthalmoscopy (AO-SLO) has recently been used to achieve exquisite subcellular resolution imaging of the mouse retina. Wavefront sensing-based AO typically restricts the field of view to a few degrees of visual angle. As a consequence the relationship between AO-SLO data and larger scale retinal structures and cellular patterns can be difficult to assess. The retinal vasculature affords a largescale 3D map on which cells and structures can be located during in vivo imaging. Phase-variance OCT (pv-OCT) can efficiently image the vasculature with near-infrared light in a label-free manner, allowing 3D vascular reconstruction with high precision. We combined widefield pv-OCT and SLO imaging with AO-SLO reflection and fluorescence imaging to localize two types of fluorescent cells within the retinal layers: GFP-expressing microglia, the resident macrophages of the retina, and GFP-expressing cone photoreceptor cells. We describe in detail a reflective afocal AO-SLO retinal imaging system designed for high resolution retinal imaging in mice. The optical performance of this instrument is compared to other state-of-the-art AO-based mouse retinal imaging systems. The spatial and temporal resolution of the new AO instrumentation was characterized with angiography of retinal capillaries, including blood-flow velocity analysis. Depth-resolved AO-SLO fluorescent images of microglia and cone photoreceptors are visualized in parallel with 469 nm and 663 nm reflectance images of the microvasculature and other structures. Additional applications of the new instrumentation are discussed.

AB - Adaptive optics scanning laser ophthalmoscopy (AO-SLO) has recently been used to achieve exquisite subcellular resolution imaging of the mouse retina. Wavefront sensing-based AO typically restricts the field of view to a few degrees of visual angle. As a consequence the relationship between AO-SLO data and larger scale retinal structures and cellular patterns can be difficult to assess. The retinal vasculature affords a largescale 3D map on which cells and structures can be located during in vivo imaging. Phase-variance OCT (pv-OCT) can efficiently image the vasculature with near-infrared light in a label-free manner, allowing 3D vascular reconstruction with high precision. We combined widefield pv-OCT and SLO imaging with AO-SLO reflection and fluorescence imaging to localize two types of fluorescent cells within the retinal layers: GFP-expressing microglia, the resident macrophages of the retina, and GFP-expressing cone photoreceptor cells. We describe in detail a reflective afocal AO-SLO retinal imaging system designed for high resolution retinal imaging in mice. The optical performance of this instrument is compared to other state-of-the-art AO-based mouse retinal imaging systems. The spatial and temporal resolution of the new AO instrumentation was characterized with angiography of retinal capillaries, including blood-flow velocity analysis. Depth-resolved AO-SLO fluorescent images of microglia and cone photoreceptors are visualized in parallel with 469 nm and 663 nm reflectance images of the microvasculature and other structures. Additional applications of the new instrumentation are discussed.

KW - Active or adaptive optics

KW - Imaging systems

KW - Ophthalmic optics and devices

KW - Ophthalmology

KW - Optical coherence tomography

KW - Visual optics, comparative animal models

UR - http://www.scopus.com/inward/record.url?scp=84945972585&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84945972585&partnerID=8YFLogxK

U2 - 10.1364/BOE.6.002191

DO - 10.1364/BOE.6.002191

M3 - Article

AN - SCOPUS:84945972585

VL - 6

SP - 2191

EP - 2210

JO - Biomedical Optics Express

JF - Biomedical Optics Express

SN - 2156-7085

IS - 6

ER -