Multiphoton versus confocal high resolution z-sectioning of enhanced green fluorescent microtubules

Increased multiphoton photobleaching within the focal plane can be compensated using a Pockels cell and dual widefield detectors

Douglas Robert Drummond, N. Carter, R. A. Cross

研究成果: ジャーナルへの寄稿記事

23 引用 (Scopus)

抄録

Multiphoton excitation was originally projected to improve live cell fluorescence imaging by minimizing photobleaching effects outside the focal plane, yet reports suggest that photobleaching within the focal plane is actually worse than with one photon excitation. We confirm that when imaging enhanced green fluorescent protein, photobleaching is indeed more acute within the multiphoton excitation volume, so that whilst fluorescence increases as predicted with the square of the excitation power, photobleaching rates increase with a higher order relationship. Crucially however, multiphoton excitation also affords unique opportunities for substantial improvements to fluorescence detection. By using a Pockels cell to minimize exposure of the specimen together with multiple nondescanned detectors we show quantitatively that for any particular bleach rate multiphoton excitation produces significantly more signal than one photon excitation confocal microscopy in high resolution Z-axis sectioning of thin samples. Both modifications are readily implemented on a commercial multiphoton microscope system.

元の言語英語
ページ(範囲)161-169
ページ数9
ジャーナルJournal of Microscopy
206
発行部数2
DOI
出版物ステータス出版済み - 5 30 2002

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Photobleaching
Microtubules
Detectors
Fluorescence
high resolution
detectors
cells
Photons
excitation
Microtomy
Imaging techniques
Confocal microscopy
Optical Imaging
fluorescence
Confocal Microscopy
Microscopes
Proteins
photons
microscopes
microscopy

All Science Journal Classification (ASJC) codes

  • Pathology and Forensic Medicine
  • Histology

これを引用

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