Micromanipulation by Light in Biology and Medicine

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ISBN: 9780817638733 | 0817638733
Cover: Hardcover
Copyright: 3/1/1999

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Institut fur Molekulare Biotechnologie, Jena, Germany. Interdisciplinary text designed to bridge the gap between those who are new to the field and the specialists in research. For physicists, chemists, biologists, and physicians. Details and data are presented in boxes and 'intermezzos' are inserted which give biological or medical information. Extended-outline format.

Preface

Introduction: The history of using light as a working tool

(6)

From photons to the laser microtools

(46)

Light

(10)

Light is a special form of electromagnetic radiation

(2)

The energy carried by light

(3)

Photons and their social behavior

(1)

The (relativistic) mass of a photon

(1)

Light pressure: Where does it come from?

(1)

Cooling of atoms and molecules by light: What optical tweezers have in common with the coldest place in the Universe

(2)

Lasers: sources of peculiar light

(11)

Laser light: Spectral purity, coherence and divergence

(2)

Principles of lasing

(1)

Inversion, three and four energy-state lasers

(1)

Pumping: The primary energy source for lasers

(1)

Laser resonators and laser modes

(3)

Changing the colors: Frequency multiplication

(1)

Real lasers

(1)

Microscopes and cameras

(11)

Different types of light microscopy

(2)

Object illumination

(1)

The magnification of a microscope

(1)

A two-lens microscope

(1)

Real microscopes

(2)

Microscope objectives

(2)

Microscope detectors

(2)

Laser microbeams and optical tweezers

(14)

Choice of the laser

(1)

Choice of microscope and objective

(2)

Building a microbeam and/or optical tweezers

(1)

The coupling unit

(2)

Mounting the parts and adjusting the lasers

(1)

``Flying spot'' - and multiple beam optical tweezers

(1)

Combination of laser microbeam and optical tweezers

(5)

Why and how light can be used as a microtool

(44)

Pulsed laser microbeams

(8)

Pulsed lasers can be focused to extreme intensities

(2)

Bringing the Sun into the lab

(3)

Why is the laser microbeam so precise: The heat is absorbed immediately

(1)

The electric fields are also impressive

(2)

Caution: X-rays

(1)

Interaction of pulsed lasers with biological matter

(6)

High intensities in medical surgery

(2)

Photons cooperate at extreme intensities

(1)

Heat shocks from a laser microbeam: Too fast to damage proteins

(2)

Working in the interior of closed objects

(2)

Interaction of optical tweezers with biological material

(8)

The wavelength makes the difference

(2)

Thermal effects in continuous laser microbeams

(1)

Interaction of NdYAG optical tweezers (1064nm) with cells

(1)

The surprise: NdYAG lasers are not the best choice

(2)

Two-photon excited fluorescence for detecting damage in IR optical traps

(2)

Light pressure: Some quantitative relationships

(6)

Calculating some surprising facts

(3)

Mie particles: The color of the target affects the light force

(1)

Light forces on very small (Rayleigh) particles

(2)

Gradient forces: Full control in three dimensions

(7)

Axial and transversal effects

(3)

Why are particles pushed towards the optical axis?

(1)

Why are particles pulled against the light pressure force?

(1)

Quantitative influence of objective illumination and of refractive indices

(2)

Unprecedented accuracy and precision: Nanometers and piconewtons

(9)

The escape force method for calibrating optical tweezers

(2)

The stiffness method

(2)

Calibration by thermal motion: The corner frequency method

(1)

Calibration by thermal motion: The minimal information method

(1)

Split photodiodes and interferometers for nanometer accuracies

(3)

From the first simple experiments to sophisticated applications of laser microtools

(32)

Microbeams in developmental biology

(9)

Drosophila: From the embryos to the organism

(2)

Laser microbeams study of the development of the roundworm Caenorhabditis elegans

101

(1)

The nervous system of Caenorhabditis elegans

102

(1)

Other nervous systems

103

(1)

Plant root development: Clonal or positional determination of cell fate?

104

(2)

Intermezzo I: Cells and cellular organelles

106

(6)

Prokaryotic cells: Bacteria

106

(1)

Cell membranes

107

(1)

Eukaryotic cells: Cells with a nucleus

107

(2)

The nucleus of eukaryotic cells

109

(1)

The cytoskelton and structure-stabilizing intracellular strands

109

(1)

Subcellular structures: Mitochondria and chloroplasts

110

(1)

Cell types for work with laser microbeams and optical tweezers

111

(1)

Laser microbeam studies on unicellular organisms, cells and subcellular structures

112

(9)

Negative and positive chemotaxis induced by laser ablation

113

(1)

Laser microbeam treatment of flagella, cilia and pseudopodia in unicellular organisms

113

(1)

Ablation of melanophores in fish

114

(1)

Ablation of mitochondria to study heart function

115

(1)

Nucleoli

116

(1)

Actomyosin fibrils and cytoplasmic strands

117

(1)

Laser ablation of interphase chromatin in the nucleus

118

(3)

Optical tweezers enter the stage

121

(8)

Optical trapping of viruses and bacteria

122

(1)

The step toward infrared optical tweezers

123

(1)

Single-cell sorters

124

(1)

Brakes for biological motors: The compliance of bacterial flagella

125

(1)

The slipper animalcule survives moving subcellular organelles and the nucleus

126

(1)

Neural growth cone membranes

126

(3)

Mitosis, molecular motors and muscles: What laser microtools can teach us

129

(30)

Intermezzo II: Reproduction and motion

129

(9)

The cell cycle

129

(2)

Details of mitosis

131

(1)

Details of the interphase

132

(1)

Microtubules

133

(1)

Intracellular traffic: cytoplasmic streaming

134

(1)

Molecular motors

135

(2)

Forces generated by single molecules

137

(1)

What drives cell division? Laser microtools help to find the answer

138

(8)

From the early days to complete micromanipulation by laser light

139

(1)

Kinetochores and centrosomes: The secret of symmetric distribution of chromosomes

139

(1)

Microablation studies on intranuclear metaphase chromosomes

140

(3)

Micotubule polymerization dynamics and flexibility

143

(3)

Molecular motors: True nanotechnology

146

(13)

Optical tweezers and single motor protein mechanics

146

(1)

Molecular force measurements in vivo: Mitochondria transport in the amoeba Reticulomyxa

147

(1)

Kinesin

147

(2)

Dynein - not as exact as kinesin

149

(2)

Myosin/actin: Single heads can do the work

151

(1)

Differences between skeletal and smooth muscle

152

(1)

Molecular bungee: Titin

153

(1)

Laser microbeams for preparing the smallest functional unit of a muscle

154

(5)

DNA as a molecular individuum

159

(46)

The ultimate DNA analytics: Single molecules

159

(13)

DNA: Astronomical dimensions and unlimited variability

160

(2)

Handling of DNA

162

(2)

Laser microdissection of single DNA molecules

164

(2)

The elasticity of a single DNA molecule

166

(2)

Two types of elasticity in DNA

168

(4)

Single molecule DNA reactions

172

(7)

From general properties to single molecule reactions

172

(2)

Cutting of an individual DNA molecule by the restricition endonucleases Apa 1

174

(3)

Transcription against a force

177

(2)

Intermezzo III: Genes, chromosomes and genetic diseases

179

(11)

Packing DNA: Chromatin

179

(1)

From chromatin to chromosomes

180

(3)

Banded chromosomes and cytogenetic nomenclature

183

(2)

Genome sizes of different organisms

185

(2)

Genetic diseases

187

(3)

Laser microdissection of chromosomes

190

(15)

The motivation for microdissection of chromosomes

190

(1)

Microdissection of chromosomes

191

(2)

Submicroscopic effects of laser microdissection

193

(1)

Combined use of laser microbeam and optical tweezers

194

(2)

Microcloning

196

(2)

PCR (Polymerase Chain Reaction)

198

(7)

From biology to medicine

205

(46)

Laser microtools in plant cell biology

205

(10)

Optical trapping in the interior of plant cells

205

(4)

Simulating microgravity in the alga Chara

209

(3)

Preparing free plant membranes for patch clamp studies

212

(3)

Microperforation of cell walls and cell membranes

215

(8)

Laser microinjection: What do biologists want to know

215

(2)

Laser-assisted gene transfer into animal cells

217

(2)

Direct microinjection of genes into plant cells: The laser is often without alternative

219

(2)

Injection of DNA into isolated chloroplasts and into chloroplasts within a plant cell

221

(2)

From laser-induced cell fusion to antibodies and immunology

223

(13)

How to avoid sex: Cell fusion in suspension

224

(1)

Fusion of plant protoplasts

225

(2)

Why single-cell fusion? Some speculation

227

(1)

Laser-induced fusion of B cells with myeloma cells

228

(2)

Increasing the laser fusion yield by chemicals

230

(1)

Increasing the yield by combining laser microbeam and optical tweezers

231

(1)

Optical tweezers-based immunosensors and detectors of virus-cell adhesion

232

(1)

Optical tweezers reveal details of the attack on the immune system on cancer

233

(3)

Towards medical diagnostics: From blood to cancer

236

(7)

Optical tweezers as wall-free microvessels to study red blood cell elasticity

236

(1)

Measuring the elasticity of red blood cells

237

(2)

Red cell adhesion: Laser tweezers test two models

239

(1)

Studies on the isolated membrane skeleton

239

(1)

Tissue microdissection in cancer research: Background

240

(1)

Laser capture microdissection and laser pressure catapulting

241

(2)

Laser microbeam and optical tweezers in reproduction medicine

243

(8)

Laser zona drilling (LZD) and handling of sperm cells by optical tweezers

244

(2)

Measuring effects of optical tweezers on sperm motility

246

(1)

Laser inactivation of extra pronuclei

247

(1)

Helping childless couples: Do laser microtools improve human IVF?

248

(1)

A word of caution

249

(2)

Appendix

251

(46)

A1 Geometrical optics

251

(10)

A1.1 From waves to rays

251

(1)

A1.2 Refraction and reflection

252

(1)

A1.3 Refraction by a spherical surface

253

(2)

A1.4 The index of refraction for selected materials

255

(1)

A1.5 Wavelength dependence on the index of refraction

256

(1)

A1.6 Reflection below the glancing angle

257

(1)

A1.7 Coating of optical elements

258

(1)

A1.8 Light guides: Simple optical instruments

259

(1)

A1.9 Prisms

260

(1)

A2 Ray optics: Lenses and image formation

261

(7)

A2.1 Lenses

261

(1)

A2.2 Different types of lenses

261

(1)

A2.3 Imaging by a simple lens

262

(2)

A2.4 Magnification and the thin lens equation

264

(2)

A2.5 The magnifying glass

266

(1)

A2.6 Aberrations, lens errors

266

(2)

A2.7 The numerical aperture of a lens

268

(1)

A3 Resolution of a microscope and focusing a laser

268

(9)

A3.1 The Bessel function and maximal focusing of a laser

268

(4)

A3.2 The resolution of a microscope

272

(1)

A3.3 Experimental determination of focal radii

273

(1)

A3.4 Visibility of objects

274

(1)

A3.5 Why lasers can be focused better than thermal light

275

(2)

A4 Light as a wave: Mathematical representation

277

(6)

A4.1 The wave equations

277

(2)

A4.2 Polarized light

279

(1)

A4.3 Interference

280

(2)

A4.4 Standing waves and modes

282

(1)

A5 Absorption, scattering and fluorescence

283

(14)

A5.1 Absorption and scattering make objects visible and allow work to be done on them

283

(2)

A5.2 Absorption and emission in a two-state atom

285

(2)

A5.3 The Lambert Beer law

287

(1)

A5.4 Two- and three-photon absorption

288

(2)

A5.5 Rayleigh scattering

290

(1)

A5.6 The detailed two energy state diagram of fluorescence

291

(1)

A5.7 Fluorescence and phosphorescence lifetimes as a basis for explaining laser action

292

(5)

Index

297

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Micromanipulation by Light in Biology and Medicine

Summary

Table of Contents

Supplemental Materials