Biology Reference
In-Depth Information
CHAPTER 9
Confocal and Multiphoton Imaging of
Intracellular Ca
2
þ
Ole Johan Kemi
*
*School of Life Sciences
University of Glasgow
United Kingdom
†
Cairn Research Limited
Faversham, Kent
United Kingdom
Abstract
I. Why Study Ca
2
þ
Signaling with Confocal and Multiphoton Microscopy
II. Confocal Microscopy
III. Limitations in Speed of Confocal Imaging
IV. Laser Scanning Confocal Microscopy
V. Total Internal Reflection Fluorescence Microscopy
VI. F
¨
rster Resonance Energy Transfer Microscopy
VII. Parallel Scanning Confocal Systems
VIII. Spinning Disk Confocal Microscopy
IX. Programmable Matrix Microscopy
X. Advantages and Disadvantages of Confocal Microscopy
XI. Multiphoton Excitation Laser Scanning Microscopy
XII. Ca
2
þ
Indicators for Use in Confocal and Multiphoton Microscopy
XIII. Use of Dyes for Single-Photon Confocal Microscopy
XIV. Use of Dyes for 2P Excitation Microscopy
XV. Is It Worth Converting the Intracellular Fluorescence Signal to [Ca
2
þ
]?
XVI. Calibration of Single Wavelength Dyes
XVII. Estimation of
F
max
Values
XVIII. Estimation of
F
min
or the Dynamic Range of the Dye
XIX. Consequence of Errors in Estimation of Intrinsic and Dye Fluorescence
XX. Multimodal and Multiple Fluorophore Confocal and Multiphoton Microscopy
References
