Techniques in Light Microscopy

Auteur
Steven E. Ruzin was Director of the CNR Biological Imaging Facility at the University of California, Berkeley from 1989 to 2022 and has been Curator of the Golub Collection of antique microscopes since 2004. He is now Emeritus Professor at the University of California, Berkley.

Texte du rabat

The book describes in a simple and readable format the way microscopes magnify images, and all of the techniques that are available today. It covers all the existing epifluorescence techniques including super-resolution and provides historical context on inventions pertaining to the development of the light microscope.

Résumé
This textbook provides an accessible and pedagogical explanation of the way microscopes magnify images and covers all techniques to date in transmitted and fluorescent light microscopy. The first section covers basic optics as it relates to microscopy. The second section describes all the major optical techniques of transmitted light microscopy, starting with brightfield, through darkfield, polarized light, phase contrast, differential interference contrast, and Hoffman modulation contrast microscopy. The final third of the book covers all the techniques of fluorescence microscopy. It begins with a simple explanation of fluorescence and is followed by Widefield epifluorescence, confocal, and 2-photon microscopy. This is followed by computational imaging including restoration ('deconvolution') microscopy, and all the present super-resolution techniques. The book concludes by discussing attainable resolution using transmitted light microscopy, including a thorough discussion of the Rayleigh, Sparrow, and other criteria of resolution, ending with a short discussion of the common pitfalls that result in decreased microscope resolution. The final chapter in the book is a short history of the microscope, beginning with the ancients, then discussing three seminal natural philosophers: Leeuwenhoek, Swammerdam, and Hooke. The remaining sections of the history chapter cover mechanical and optical advancements in the history of transmitted light microscopy. Finally, it includes a short history of fluorescence microscopy starting with Köhler and Rohr's first use of fluorescence microscopy in 1905, and ending with a description of the Sarastro Phoibos 1000, the first commercial confocal microscope.

Contenu

• Part I

• 1: Light Microscope Optics

• 2: Lens Aberrations

• 3: The Compound Microscope

• Part II

• 4: Transmitted Light Techniques

• 5: Polarized Light Microscopy

• 6: Phase Contrast Microscopy

• 7: Differential Interference Contrast Microscopy

• Part III

• 8: Epifluorescence Microscopy

• 9: Selective/Single Plane Imaging Microscopy

• 10: Confocal Microscopy

• 11: Two-Photon Microscopy

• 12: Restoration Microscopy

• 13: Super-Resolution Microscopy

• 14: Resolution

• 15: A Brief History of the Light Microscope

• Appendix I: Digital imaging and sampling

• Appendix II: Synopsis of transmitted light microscopy techniques

• Appendix III: Synopsis of epifluorescence microscopy techniques