Methods in Cell Biology

Volume 55 in Methods in Cell Biology is a concise laboratory book that emphasizes the methods and technologies needed to use single polarized laser light source that functions simultaneously as an optical trap and a dual-beam interferometer. * * Provides a practical laboratory guide for methods and technologies used with laser tweezers * Includes comprehensive and easy-to-follow protocols

Contributors

ix

Preface

xi

Forces of a Single-Beam Gradient Laser Trap on a Dielectric Sphere in the Ray Optics Regime

A. Ashkin

Introduction

1

(3)

Light Forces in the Ray Optics Regime

4

(5)

Force of the Gradient Trap on Spheres

9

(7)

Effect of Mode Profiles and Index of Refraction on Trapping Forces

16

(4)

Concluding Remarks

20

(9)

Appendix I: Force of a Ray on a Dielectric Sphere

21

(1)

Appendix II: Force on a Sphere for Trap Focus along Y Axis

22

(1)

Appendix III: Force on a Sphere for an Arbitrarily Located Trap Focus

23

(2)

References

25

(4)

Basic Laser Tweezers

Ronald E. Sterba

Michael P. Sheetz

General Description

29

(1)

Microscope

30

(3)

Laser

33

(2)

Optics and Layout

35

(1)

System Setup

36

(1)

Alignment

37

(1)

Translation

38

(1)

Video Recording and Analysis

38

(1)

Accessories

39

(1)

Summary

40

(3)

Appendix: Laser Tweezers Parts List

40

(1)

References

41

(2)

A Simple Assay for Local Heating by Optical Tweezers

Scot C. Kuo

Introduction

43

(1)

Methods

44

(1)

Results

44

(3)

References

45

(2)

Reflections of a Lucid Dreamer: Optical Trap Design Considerations

Amit D. Mehta

Jeffrey T. Finer

James A. Spudich

Introduction

47

(1)

Choice of Trapping Laser

48

(2)

Optical Layout

50

(3)

Imaging

53

(3)

High-Resolution Position Measurement

56

(2)

Noise Sources

58

(4)

Feedback

62

(3)

Calibration

65

(2)

Analysis

67

(1)

Conclusion

68

(3)

References

68

(3)

Laser Scissors and Tweezers

Michael W. Berns

Yona Tadir

Hong Liang

Bruce Tromberg

Introduction

71

(3)

Mechanisms of Interaction

74

(7)

Biological Studies

81

(13)

Summary

94

(5)

References

94

(5)

Optical Force Microscopy

Andrea L. Stout

Watt W. Webb

Introduction

99

(1)

AFM-like Applications

100

(5)

Experimental Design

105

(8)

New Directions

113

(4)

References

115

(2)

Single Molecule Imaging and Nanomanipulation of Biomolecules

Yoshie Harada

Takashi Funatsu

Makio Tokunaga

Kiwamu Saito

Hideo Higuchi

Yoshiharu Ishii

Toshio Yanagida

Introduction

117

(1)

Visualization of Single Fluorophores in Aqueous Solution

118

(5)

Application

123

(4)

Perspectives

127

(2)

References

128

(1)

Signals and Noise in Micromechanical Measurements

Frederick Gittes

Christoph F. Schmidt

Introduction

129

(2)

Spectral Data Analysis

131

(5)

Brownian Motion of a Harmonically Bound Particle

136

(4)

Noise Limitations on Micromechanical Experiments

140

(7)

Sources of Instrumental Noise

147

(7)

Conclusions

154

(3)

References

154

(3)

Cell Membrane Mechanics

Jianwu Dai

Michael P. Sheetz

Perspectives and Overview

157

(3)

Laser Optical Tweezers

160

(1)

Bead Coating

160

(1)

Calibration of the Laser Tweezers

161

(4)

Tracking the Bead Position

165

(1)

Memberane Tether Formation and Tether Force Measurement

166

(1)

Membrane Tether Force and Membrane Mechanical Properties

167

(1)

Membrane Tension and Its Significance

168

(6)

References

170

(4)

Application of Laser Tweezers to Studies of the Fences and Tethers of the Membrane Skeleton that Regulate the Movements of Plasma Membrane Proteins

Akihiro Kusumi

Yasushi Sako

Takahiro Fujiwara

Michio Tomishige

Introduction

174

(2)

Studying Various Modes of Protein Motion in the Plasma Membrane Using Single-Particle Tracking

176

(1)

Technical Details for SPT

176

(3)

Instrumental Setup and Calibration of Laser Tweezers

179

(2)

Dragging Receptor Molecules along the Plasma Membrane and Finding Obstacles in the Path

181

(4)

Laser Tweezers Together with SPT Can Reveal the Presence of Elastic Intercompartmental Barriers

185

(3)

Studying the Fence and Tether Effects of the Membrane Skeleton Using Laser Tweezers

188

(4)

SPT and Laser Tweezers Are Opening Up Possibilities for Scientists to Learn about the Molecular Mechanics in Cells by Directly Handling Single Molecules in Living Cells

192

(3)

References

192

(3)

In Vivo Manipulation of Internal Cell Organelles

Harald Felgner

Franz Grolig

Otto Muller

Manfred Schliwa

Introduction

195

(1)

Optical Tweezers Setup

196

(1)

Reticulomyxa: Organelle Movement and Artificial Membrane Tubes

197

(2)

Spirogyra: Cytoplasmic Streaming

199

(6)

References

202

(3)

Optical Chopsticks: Digital Synthesis of Multiple Optical Traps

Justin E. Molloy

Introduction

205

(1)

Trapping Configurations

206

(2)

Mechanical Deflectors (Mirrors)

208

(1)

Acousto-optic Deflectors

208

(2)

Position: Control and Noise

210

(2)

Computer Control of Trap Positions

212

(1)

Other Developments

213

(4)

References

215

(2)

Index

217

(6)

Volumes in Series

223

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Methods in Cell Biology

Summary

Table of Contents

Supplemental Materials