4. Microscopy

High-power objectives

High-power objectives pose several design problems. Because the focal length of an objective decreases as the N.A. and magnifying power increase, the working distance, or distance from the front of the objective to the top of the slide, is shorter for higher-power objectives. The need to use additional elements in the lens system for high magnifications further shortens the working distance to only 10 to 20 percent of the focal length. Thus, a 40× objective of 4-mm (0.2-inch) focal length may have a working distance of less than 0.4 mm (0.02 inch), so objectives with an increased working distance have been designed. These use a negative lens element between the object and the eyepiece, which has the added attraction of providing some field flattening as well. These objectives are especially of value in use with video systems.

In objectives with magnifying powers of 25× or greater, meniscus-shaped aplanatic elements are designed into the microscope objective in the space between the object and the pairs of doublets that carry out the relayed imaging. These aplanatic components have the property of converging the light without adding spherical aberration to the image and provide an increase in the N.A. without introducing significant aberration.

The highest-power microscope objective available is the immersion objective. When this type of objective is used, a drop of oil must be placed between the object on the microscope slide and the objective. The oil used has an R.I. that matches that of the glass in the first component of the objective.

The first component of immersion objectives is generally a hyper-hemisphere (a small optical surface shaped like a hemisphere but with a boundary curve exceeding 180°), which acts as an aplanatic coupler between the slide and the rest of the microscope objective. An immersion objective with a high N.A. typically consists of a hyper-hemisphere followed by one or two aplanatic collectors and then two or more sets of doublets. Such objectives are made with magnifying powers greater than 50×, the extreme being about 100×.

Water-immersion lenses are also available. These use water as an immersion liquid and allow biologists to examine specimens in a watery medium without the burden of a cover slip confining the living organisms.

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