Confocal and multiphoton microscopy for imaging at depth in living tissue

J.M. Girkin, A.J. Wright, S. Poland, B.A. Patterson, M.J. Padgett

Research output: Contribution to journalArticle

Abstract

The desire to image with sub micron resolution at ever increasing depths into living samples is providing optical physicists with the latest in a long line of challenges presented by life science researchers. The advent of confocal, and subsequently multiphoton microscopy, has opened up exciting new possibilities but simultaneously posed new challenges. As one images ever more deeply into the sample, the optical properties of the tissue distort the image significantly lowering the resolution and, in the case of multiphoton imaging in particular, decreasing the fluorescence yield as the excitation volume rises. The recent use of active optical elements has shown a way forward in restoring high contrast high resolution images at depth. However, significant issues on the actual shape required on such an element are as yet unresolved. We report on two recent advances in this area. The first is the use of a range of optimisation algorithms to restore the optical point spread function and hence improve the image quality at depth. The second is a radically new approach incorporating two active elements, a slow spatial light modulator and a fast deformable mirror, to actively lock up the system. We report on the latest advances in active image compensation where conections at over 5OOmicrons into the sample have been made using a combination of deformable mirrors and spatial light modulators.
Original languageEnglish
Pages (from-to)2P1 - 2P8
JournalMultiphoton Microscopy in the Biomedical Sciences IV (Proceedings of SPIE)
Volume6047
Issue number2
DOIs
Publication statusPublished - 27 Oct 2006

Fingerprint

Multiphoton Microscopy
Confocal Microscopy
Microscopic examination
Imaging
Tissue
microscopy
Imaging techniques
Light
Biological Science Disciplines
Optical transfer function
Image resolution
Optical devices
Spatial Light Modulator
Deformable Mirror
Image quality
Microscopy
Mirrors
Optical properties
deformable mirrors
Fluorescence

Keywords

  • optical physics
  • confocal microscopy

Cite this

Girkin, J.M. ; Wright, A.J. ; Poland, S. ; Patterson, B.A. ; Padgett, M.J. / Confocal and multiphoton microscopy for imaging at depth in living tissue. In: Multiphoton Microscopy in the Biomedical Sciences IV (Proceedings of SPIE). 2006 ; Vol. 6047, No. 2. pp. 2P1 - 2P8.
@article{0ae4e234a13b42f9912b9a40193be5b2,
title = "Confocal and multiphoton microscopy for imaging at depth in living tissue",
abstract = "The desire to image with sub micron resolution at ever increasing depths into living samples is providing optical physicists with the latest in a long line of challenges presented by life science researchers. The advent of confocal, and subsequently multiphoton microscopy, has opened up exciting new possibilities but simultaneously posed new challenges. As one images ever more deeply into the sample, the optical properties of the tissue distort the image significantly lowering the resolution and, in the case of multiphoton imaging in particular, decreasing the fluorescence yield as the excitation volume rises. The recent use of active optical elements has shown a way forward in restoring high contrast high resolution images at depth. However, significant issues on the actual shape required on such an element are as yet unresolved. We report on two recent advances in this area. The first is the use of a range of optimisation algorithms to restore the optical point spread function and hence improve the image quality at depth. The second is a radically new approach incorporating two active elements, a slow spatial light modulator and a fast deformable mirror, to actively lock up the system. We report on the latest advances in active image compensation where conections at over 5OOmicrons into the sample have been made using a combination of deformable mirrors and spatial light modulators.",
keywords = "optical physics, confocal microscopy",
author = "J.M. Girkin and A.J. Wright and S. Poland and B.A. Patterson and M.J. Padgett",
note = "Special issue entitled: Fourth International Conference on Photonics and Imaging in Biology and Medicine.",
year = "2006",
month = "10",
day = "27",
doi = "10.1117/12.710985",
language = "English",
volume = "6047",
pages = "2P1 -- 2P8",
journal = "Multiphoton Microscopy in the Biomedical Sciences IV (Proceedings of SPIE)",
issn = "1605-7422",
number = "2",

}

Confocal and multiphoton microscopy for imaging at depth in living tissue. / Girkin, J.M.; Wright, A.J.; Poland, S.; Patterson, B.A.; Padgett, M.J.

In: Multiphoton Microscopy in the Biomedical Sciences IV (Proceedings of SPIE), Vol. 6047, No. 2, 27.10.2006, p. 2P1 - 2P8.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Confocal and multiphoton microscopy for imaging at depth in living tissue

AU - Girkin, J.M.

AU - Wright, A.J.

AU - Poland, S.

AU - Patterson, B.A.

AU - Padgett, M.J.

N1 - Special issue entitled: Fourth International Conference on Photonics and Imaging in Biology and Medicine.

PY - 2006/10/27

Y1 - 2006/10/27

N2 - The desire to image with sub micron resolution at ever increasing depths into living samples is providing optical physicists with the latest in a long line of challenges presented by life science researchers. The advent of confocal, and subsequently multiphoton microscopy, has opened up exciting new possibilities but simultaneously posed new challenges. As one images ever more deeply into the sample, the optical properties of the tissue distort the image significantly lowering the resolution and, in the case of multiphoton imaging in particular, decreasing the fluorescence yield as the excitation volume rises. The recent use of active optical elements has shown a way forward in restoring high contrast high resolution images at depth. However, significant issues on the actual shape required on such an element are as yet unresolved. We report on two recent advances in this area. The first is the use of a range of optimisation algorithms to restore the optical point spread function and hence improve the image quality at depth. The second is a radically new approach incorporating two active elements, a slow spatial light modulator and a fast deformable mirror, to actively lock up the system. We report on the latest advances in active image compensation where conections at over 5OOmicrons into the sample have been made using a combination of deformable mirrors and spatial light modulators.

AB - The desire to image with sub micron resolution at ever increasing depths into living samples is providing optical physicists with the latest in a long line of challenges presented by life science researchers. The advent of confocal, and subsequently multiphoton microscopy, has opened up exciting new possibilities but simultaneously posed new challenges. As one images ever more deeply into the sample, the optical properties of the tissue distort the image significantly lowering the resolution and, in the case of multiphoton imaging in particular, decreasing the fluorescence yield as the excitation volume rises. The recent use of active optical elements has shown a way forward in restoring high contrast high resolution images at depth. However, significant issues on the actual shape required on such an element are as yet unresolved. We report on two recent advances in this area. The first is the use of a range of optimisation algorithms to restore the optical point spread function and hence improve the image quality at depth. The second is a radically new approach incorporating two active elements, a slow spatial light modulator and a fast deformable mirror, to actively lock up the system. We report on the latest advances in active image compensation where conections at over 5OOmicrons into the sample have been made using a combination of deformable mirrors and spatial light modulators.

KW - optical physics

KW - confocal microscopy

UR - http://dx.doi.org/10.1117/12.710985

U2 - 10.1117/12.710985

DO - 10.1117/12.710985

M3 - Article

VL - 6047

SP - 2P1 - 2P8

JO - Multiphoton Microscopy in the Biomedical Sciences IV (Proceedings of SPIE)

JF - Multiphoton Microscopy in the Biomedical Sciences IV (Proceedings of SPIE)

SN - 1605-7422

IS - 2

ER -