Aberration correction for confocal imaging in refractive‐index‐mismatched media
Abstract
Summary: A major limitation to the use of confocal microscopes to image thick biological tissue lies in the dramatic reduction in both signal level and resolution when focusing deep into a refractive‐index‐mismatched specimen. This limitation may be overcome by measuring the wavefront aberration and pre‐shaping the input beam so as to cancel the effects of aberration. We consider the images of planar and point objects in brightfield, single‐photon fluorescence and two‐photon fluorescence imaging. In all cases, the specimens are imaged using an oil‐immersion objective through various thicknesses of water. The question of finite‐sized pinhole is addressed and it is found, in general, that it is sufficient to correct only the first two or three orders of spherical aberration to restore adequate image signal level and optical resolution, at imaging depths of up to 50‐100 wavelengths.
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A detection numerical aperture (NA) of 0.6 is used; lower NA values do not sufficiently resolve signals from adjacent voxels, whereas higher NA values introduce more significant spherical aberration with depth.
“A detection numerical aperture (NA) of 0.6 is used; lower NA values do not sufficiently resolve signals from adjacent voxels, whereas higher NA values introduce more significant spherical aberration with depth34–36.”
From Laser writing in glass for dense, fast and efficient archival data storage (2026) · cites this paper as [35]
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Two-photon excitation microscopy for the study of living cells and tissues
Benninger, R. K. P., & Piston, D. W. (2013) · Current Protocols in Cell Biology · doi:10.1002/0471143030.cb0411s59 · Cited by 311 · see citing works
Adaptive Optical Two-Photon Microscopy for Surface-Profiled Living Biological Specimens
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Laser writing in glass for dense, fast and efficient archival data storage
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- Authors
- Booth, M. J., Neil, M. A. A., & Wilson, T.
- Affiliations
- University of Oxford (GB)
- Venue
- Journal of Microscopy
- Publisher
- Wiley
- Published
- 1998-11
- Volume
- 192
- Issue
- 2
- Pages
- 90-98
- ISSN
- 0022-2720 (ISSN-L)
- 1365-2818
- OpenAlex
- W2083793763
- Cited by
- 248
- References
- 6
- Citations by year
- 2026: 11 · 2025: 6 · 2024: 13 · 2023: 11 · 2022: 8 · 2021: 8 · 2020: 16 · 2019: 8 · 2018: 16 · 2017: 9 · 2016: 10 · 2015: 13 · 2014: 16 · 2013: 19 · 2012: 11
- FWCI
- 1.88
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Journal of Microscopy (publishedVersion)
- Funders
- Biotechnology and Biological Sciences Research Council
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Biophysics › Advanced Fluorescence Microscopy Techniques
- Physical Sciences › Physics and Astronomy › Atomic and Molecular Physics, and Optics › Digital Holography and Microscopy
- Physical Sciences › Physics and Astronomy › Atomic and Molecular Physics, and Optics › Adaptive optics and wavefront sensing
- Keywords
- aberration correction, confocal microscopy, refractive index mismatch, wavefront aberrations, spherical aberration, two-photon fluorescence, single-photon fluorescence, oil immersion objective, water immersion, pinhole size, image depth, optical resolution
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Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
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