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Spie Press Book

Optical Design of Microscopes
Author(s): George H. Seward
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Book Description

Knowledge of microscope design is rapidly becoming more important. Microscopes are used in critical applications such as drug development, clinical tests, and genomics. Considerable expertise is required for the evaluation, design, and manufacture of these instruments. Several subsystems must be integrated: the source, the illumination optics, the specimen, the objective lens, the tube optics, and the sensor. The large numerical aperture of a microscope is essential for small spot size and high brightness; however, the large numerical aperture also presents difficult issues in optical design and fabrication.

This book provides a foundation for developing design expertise through education, practice, and exploration. It is suitable for lens designers, optical engineers, and students with a basic knowledge of microscope structure.

Book Details

Date Published: 30 April 2010
Pages: 258
ISBN: 9780819480958
Volume: TT88

Table of Contents
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Chapter 1 Optical Design Concepts
1.1 A Value Proposition
1.2 Specimen Model
1.3 Detector Parameters
1.4 Numerical Aperture (NA)
1.5 Wave Propagation
1.6 Geometric Aberrations
1.7 Image Contrast
1.8 Manufacturing
1.9 Assembly
Chapter 2 Basic Microscope Concepts
2.1 Magnification
2.3 Finite Tube Length
2.4 ∞-Corrected Objective
2.5 Tube Lens
2.6 Ocular Lens
2.7 Refractive Objects
2.8 Diffractive Objects
2.9 Dark Field
Chapter 3 Geometric Optics
3.1 Ray Tracing
3.2 Cardinal points
3.3 Stops
3.4 Gaussian Lens Formula
3.5 Image Types
3.6 Optical Power
3.7 Paraxial Optics
3.8 Relay Lens
3.9 Magnifier
Chapter 4 Aberrations
4.1 Seidel Aberrations
4.2 Chromatic Aberrations
4.3 Other Aberrations
4.4 Aspheric Surfaces
Chapter 5 Basic Physical Optics
5.1 Importance of Physical Optics
5.2 Wave Equation
5.3 Refractive Index
5.4 Dispersion
5.5 Refraction and Reflection
5.6 Emission
5.7 Absorption
5.8 Evanescent Field
5.9 Space-Angle Product
5.10 Coherence
5.11 Airy Pattern
5.12 Gaussian Beam Propagation
5.13 Transfer Functions
5.14 Gaussian Estimate of Airy Pattern
5.15 Scatter
5.16 Interference Filters
Chapter 6 Fluorescence
6.1 Absorption Parameters
6.2 Electron States
6.3 Energy Diagrams
6.4 Fluorophores
Chapter 7 Optical Design Metrics
7.1 CAD Tools
7.2 Wavefront Error
7.3 Ray-Intercept Plot
7.4 Spot Diagram
7.5 Point-Spread Plot
7.6 Encircled Energy Plot
7.7 Modulation transfer function
7.8 Edge Spread
7.9 Lens Report
7.10 Relative Illumination
7.11 Surface Form Error
7.12 Manufacturing Standards
Chapter 8 Image Contrast
8.1 Radiometry
8.2 Expression of Contrast
8.3 Shot Noise
8.4 Emittance Patterns
8.5 Angular Collection Efficiency
8.6 Spatial Collection Efficiency
8.7 Full-Pixel Contrast
8.8 Subpixel Contrast
8.9 Point-Source Contrast
8.10 Full-Pixel Airy Contrast
Chapter 9 Microlens Formats
9.1 10XR Double Gauss
9.2 10XR Microlens
9.3 2XR Microlens
9.4 1X Microlens
9.5 2XR Telecentric Spectroscopy Lens
Chapter 10 Illumination Systems
10.1 Condenser
10.2 Abbe illumination
10.3 Nelson illumination
10.4 Diffusers
10.5 Kohler Illumination
10.6 Matched Stops
10.7 Light-Emitting Diodes (LEDs)
10.8 Aspheric Plus Singlet Relay
10.9 Achromatic Aspheric Plus Doublet Relay
10.10 Abbe Condenser
10.11 Abbe Aspheric
10.12 Total Internal Reflection Fluorescence (TIRF) Illumination
Chapter 11 Cover Strata
11.1 Importance of Specimen Tolerance
11.2 Perfect 10X for Air
11.3 10X Objective with Cover Glass in Place of Air
11.4 10X Objective with Microscope Slide in Place of Air
11.5 40X Objective with Silica Cover in Place of Glass
11.6 40X Objective with Tilted Cover Glass
11.7 60X Objective with Silica Cover in Place of Glass
11.8 Strehl Versus Optical Path Length
Chapter 12 Objective Lenses
12.1 Formats
12.2 Aplanatic Surface
12.3 10X Plan Achromat
12.3 40X Fluor
12.4 60X ImmersionTIRF
12.5 100X Aplanat
12.6 10X Schwarzschild
12.7 20X Internal Parabola
Chapter 13 Tube Elements
13.1 Doublet Tube Lens
13.2 Doublet-Pair Tube Lens
13.3 Filter Types
13.4 Filter within Finite Conjugate Distance
13.5 Warped Filter within an Infinity Correction
Chapter 14 Ocular Lens
14.1 Eyepiece
14.2 Pupils
14.3 Kellner Ocular
14.4 Ploessl Ocular
14.5 Erfle Ocular
Chapter 15 Sensors
15.1 CCD Sensors
15.2 Active Pixel Sensors
15.3 Photomultiplier Tubes
15.4 Film
Chapter 16 Human Vision
16.1 Physiology
16.2 Contrast Sensitivity Function
16.3 Point Spread of Lens
16.4 Lateral Inhibition of the Retina
16.5 Temporal Feedback of Photoreceptors
16.6 Saccation Point Spread
16.7 Vision Research
16.8 Temporal CSF
Chapter 17 Optical Materials
17.1 Glass Types
17.2 Glass Map
17.4 Fluorite
17.5 Short Flint
17.6 Anomalous Dispersion
17.7 Sellmeier Formula
17.8 Environmentally Safe Glass
17.9 Glass Code
17.10 Spectral Lines
17.11 Cost of Optics
17.12 Structural Materials
Chapter 18 Composition and Spectra of Materials
18.1 Glass Structure
18.2 Crown
18.3 Flint
18.4 Long Crown
18.5 Short Flint
18.6 Short-Flint Special
18.7 Environmentally Safe Short Flint
18.7 Dense Flint
Chapter 19 Advanced Concepts
19.1 Wave Equation
19.3 Refractive Index
19.4 Relative Partial Dispersion
19.5 Emission
19.6 Coherence
19.7 Gaussian Beam Power
19.8 Transfer Functions
19.9 Scatter
19.10 Interference Filters
19.11 Shot Noise
Appendix: Prescriptions
Works Consulted
Recommended Reading for an Optical Engineer


The book provides an introduction to optical design as it pertains to microscopes. The large numerical aperture (NA) of a microscope creates issues that are not present in systems such as telescopes and cameras. The importance of microscope design is growing rapidly in 2009.

Microscopes are frequently employed in drug development, clinical tests, and genomics. However, one system does not fit all applications. Considerable expertise is required for evaluation, design, and manufacture of these instruments. An integrated relationship must be established between several subsystems: the source, the illumination optics, the specimen, the objective lens, the tube optics, and the sensor.

The optical prescriptions are specified with glass name, refractive index, and Abbe number. Methods for color correction are described with specific materials. The anomalous partial dispersion in blue is described for a long crown and short flint. The origin of dispersion is related to glass composition.

Several essential topics in optics are reviewed. Geometric optics provides a simplistic description of ray tracing. Physical optics is described in two chapters: a first chapter describes the basic concepts of wave propagation with simple algebra, and a second chapter describes more advanced concepts with vector calculus. Optical aberrations describe the departure from perfection. Expressions for image contrast are defined for practical application to imaging systems. Expertise in these topics is essential for proper design of an optical instrument.

Numerous chapters rely on simple plots and basic algebra. A novice designer should easily comprehend the majority of chapters. However, a few chapters require comprehension of calculus, vector operations, and Fourier analysis. An expert in design should seek maximum comprehension of the chapter on advanced concepts at the end of the book.

This book provides a foundation for the development of expertise in optical design. Expertise is developed through education, practice, and exploration.

George H. Seward
January 2010

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