GO TO THE FULL PROTOCOL:
PDF or HTML at Wiley Online Library
Microscope Objectives
Publication Name:
Current Protocols in Cytometry
Unit Number:
Unit 2.2
DOI:
10.1002/0471142956.cy0202s58
Online Posting Date:
October, 2011
Abstract
The objective is the most crucial image-forming component of a microscope. A knowledge of the many types of objectives available and their characteristics is critical to the selection of appropriate objectives for image cytometry. This unit discusses aberrations in image formation and their correction, construction, and types of objectives, and objectives for other microscopy applications, explaining the advantages and limitations of each one. Curr. Protoc. Cytom. 58:2.2.1-2.2.15. © 2011 by John Wiley & Sons, Inc.
Keywords: imaging and microscopy; confocal microscopy; light microscopy
Table of Contents
Figures
-
Figure 2.2.1 Schematic diagram of a compound microscope, illustrating the site of intermediate image formation at the plane of the eyepiece diaphragm.
(Source: Reproduced from Abramowitz ( -
Figure 2.2.2 Formation of a real image of a point by an ideal lens.
(Source: Reproduced from Abramowitz ( -
Figure 2.2.3 Airy disk diffraction image of a point, illustrating the formation of the image of a specimen point as a disk of light rather than a dimensionless fine point.
(Source: Reproduced from Abramowitz ( -
Figure 2.2.4 Chromatic aberration in an uncorrected lens, illustrating the existence of different focal planes for different wavelengths of light.
(Source: Reproduced from Abramowitz ( -
Figure 2.2.5 Spherical aberration, or the difference of focus for light rays traversing the outer zones of a lens compared to light passing more centrally.
(Source: Reproduced from Abramowitz ( -
Figure 2.2.6 Diagrammatic representation showing the comet-like smearing of an image point from a lens that has not been corrected for coma aberration.
(Source: Reproduced from Abramowitz ( -
Figure 2.2.7 Astigmatism, an aberration in the outer lens zones caused by unequal magnification in the different azimuths. A specimen point appears in the image not as a point but as a line.
(Source: Reproduced from Abramowitz ( -
Figure 2.2.8 Barrel and pincushion distortion of parallel lines in an uncorrected lens.
(Source: Reproduced from Abramowitz ( -
Figure 2.2.9 Curvature of field, in which the image of a flat specimen is curved and therefore not in a uniform plane of focus.
(Source: Reproduced from Abramowitz ( -
Figure 2.2.10 Illustration of the light gathering ability of three lenses with increasing numerical aperture (NA), where n is the refractive index of the medium between the objective and the coverslip and µ is the half-angle of the cone of light captured by the objective.
(Source: Reproduced from Abramowitz ( -
Figure 2.2.11 Schematic, longitudinal section through a 60× Plan Apo objective lens, illustrating the numerous and complex elements of a highly corrected objective.
(Source: Diagram reprinted courtesy of Olympus America.) -
Figure 2.2.12 Diagrammatic representation of typical markings found on the case of an objective lens. -
Figure 2.2.13 Diagrammatic representation of an infinity-corrected objective system. The diagram illustrates parallel beams emerging from different azimuths (shaded) of an infinity-corrected objective being brought to focus by a tube lens.
(Source: Reproduced from Abramowitz ( -
Figure 2.2.14 Infinity space: the distance between the back of an infinity-corrected objective and the tube lens (schematic).
(Source: Reproduced from Abramowitz ( -
Figure 2.2.15 Objective system of finite tube length, showing the projection of the image by a finite objective to the intermediate image plane within the eyepiece tube.
(Source: Reproduced from Abramowitz (
Videos
Literature Cited
| Literature Cited | |
| Abramowitz, M. 1994. Optics: A Primer. Olympus America Inc., New York. | |
| Key References | |
| Abramowitz, M. 1985. Microscope Basics and Beyond. Olympus Corporation, New York. | |
| Abramowitz, M. 1987. Contrast Methods in Microscopy: Transmitted Light. Olympus Corporation, New York. | |
| Abramowitz, M. 1993. Fluorescence Microscopy: The Essentials. Olympus America Inc., New York. | |
|
Abramowitz,
1994.
See | |
| Bradbury, S. 1984. An Introduction to the Optical Microscope. Oxford University Press, Oxford, UK. | |
| Delly, J.G. 1988. Photography Through The Microscope. Eastman Kodak, Rochester, N.Y. | |
| Inoue, S. 1986. Video Microscopy. Plenum Press, New York. | |
| Leitz, E. 1938. The Microscope And Its Application. Ernst Leitz, Wetzlar, Germany. | |
| Mollring, F.K. 1976. Microscopy From The Very Beginning. Carl Zeiss, Oberkochen, Germany. | |
| Spencer, M. 1982. Fundamentals of Light Microscopy. Cambridge University Press, Cambridge, U.K. | |
| Internet Resources | |
| http://www.microscopyu.com | |
|
Web site featuring technical support and timely information about all aspects of optical microscopy. | |
