The White Confocal Microscopic Optical Sectioning in all Colors

This book offers a comprehensive introduction to confocal microscopy – with a particular focus on spectral confocal microscopy. Beginning with an introduction to optical lenses, it provides a guide to compound microscopes and explains related topics like microscopic resolution. It then presents an outline of fluorescence and its corresponding implications for microscopy. The following excursus on the confocal beam paths includes implementation of acousto-optical devices and modern sensor techniques. Complex relationships are explained in a comprehensible manner, supported by many graphical figures.

Discussing the principles of magnifying optics and the technical fundamentals and modes of operation of modern laser scanning microscopes, it is a valuable resource for student and lab technicians as well as faculty members.

The White Confocal

0          Preface          

1          Microscopy – an introduction       

1.1       Lenses

1.2       The microscope

1.2.1   The objective lens

1.2.3   The compound microscope

1.3       Resolution and the limits

1.3.1   Abbes formula

1.3.2   Glowing spots

1.3.3   Full width half maximum

1.3.4   Which ist he truth?

1.4       Beyond the limit of resolution

2          Fluorescence

2.1       What is fluorescence

2.1.1   The fluorescence process

2.1.2   Color games

2.1.3   Life times

2.2       Microscopy by fluorescence

2.2.1   Power ratio of excitation and emission

2.2.2   Transmitted light and incident light

2.2.3   Illumination

2.2.4   Excitation filter

2.2.5   Incident light and beam splitter

2.3       Artificial colours

3          Confocal microscopy

3.1       The reason

3.2       The principle

3.2.1   Spot illumination

3.3       The scanned image

3.3.1   Recording

3.3.2   Scanning procedures

3.3.3   Slice thickness

3.3.4   The third dimension

3.4       Two photon microscopy

3.5       Elements of a confocal microscope

3.5.1   Light source

3.5.2   Excitation filter

3.5.3   Primary splitter

3.5.4   Beam scanner

3.5.5   Objective lens

3.5.7   Emission filter

3.5.8   Sensor

3.6       The „white“ confocal microscope

4          Light sources

4.1       Laser

4.2       Laser types

4.3       White light laser

5          Acousto optical excitation filter

5.1       How works an acoustooptical tunable filter?

5.2       Multichannel dimmer for laser light

5.3       Spectral freedom: AOTF and white light laser

6          White beam splitting

6.1       Acoustooptical beam splitter

6.2       AOBS and white source

7          Split of the emissions

7.1       Prism

7.2       Grating

7.3       Continuous separation: the spectrum

8          Emission filtering

8.1       The line array detector

8.2       The multiband detector

9          Separation in time domain

9.1       Sensors sense photons

9.1.1   Photoelectron multiplier tube

9.1.2   Avalanche photodiode

9.1.3   Hybrid detector

9.2       White fluorescence measurement: FLIM

9.3       A white filter with high selectivity

10        Further reading

Dr. Rolf Theodor Borlinghaus, born 1956, study of biology in Konstanz and promotion to Dr. rer. nat. at the biophysical institute. From 1990 product manager for complex fluorescence microscopy and confocal microscopy with Carl Zeiss Oberkochen (later Jena). Since 1997 with Leica Lasertechnik Heidelberg, now Leica Microsystems Mannheim. Today responsible for technology transfer and training.