III.

Special-Purpose Optical Microscopes

Different microscopes have been developed for specialized uses. The stereoscopic microscope, two low-powered microscopes arranged to converge on a single specimen, provides a three-dimensional image.

The petrographic microscope is used to analyze igneous and metamorphic rock. A Nicol prism or other polarizing device polarizes the light that passes through the specimen. Another Nicol prism or analyzer determines the polarization of the light after it has passed through the specimen. Rotating the stage causes changes in the polarization of light that can be measured and used to identify and estimate the mineral components of the rock.

The dark-field microscope employs a hollow, extremely intense cone of light concentrated on the specimen. The field of view of the objective lens lies in the hollow, dark portion of the cone and picks up only scattered light from the object. The clear portions of the specimen appear as a dark background, and the minute objects under study glow brightly against the dark field. This form of illumination is useful for transparent, unstained biological material and for minute objects that cannot be seen in normal illumination under the microscope.

The phase microscope also illuminates the specimen with a hollow cone of light. However, the cone of light is narrower and enters the field of view of the objective lens. Within the objective lens is a ring-shaped device that reduces the intensity of the light and introduces a phase shift of a quarter of a wavelength. This illumination causes minute variations of refractive index in a transparent specimen to become visible. This type of microscope is particularly effective for studying living tissue.

A typical optical microscope cannot resolve images smaller than the wavelength of light used to illuminate the specimen. An ultraviolet microscope uses the shorter wavelengths of the ultraviolet region of the light spectrum to increase resolution or to emphasize details by selective absorption (see Ultraviolet Radiation). Glass does not transmit the shorter wavelengths of ultraviolet light, so the optics in an ultraviolet microscope are usually quartz, fluorite, or aluminized-mirror systems. Ultraviolet radiation is invisible to human eyes, so the image must be made visible through phosphorescence (see Luminescence), photography, or electronic scanning.

The near-field microscope is an advanced optical microscope that is able to resolve details slightly smaller than the wavelength of visible light. This high resolution is achieved by passing a light beam through a tiny hole at a distance from the specimen of only about half the diameter of the hole. The light is played across the specimen until an entire image is obtained.

The magnifying power of a typical optical microscope is limited by the wavelengths of visible light. Details cannot be resolved that are smaller than these wavelengths. To overcome this limitation, the scanning interferometric apertureless microscope (SIAM) was developed. SIAM uses a silicon probe with a tip one nanometer (1 billionth of a meter) wide. This probe vibrates 200,000 times a second and scatters a portion of the light passing through an observed sample. The scattered light is then recombined with the unscattered light to produce an interference pattern that reveals minute details of the sample. The SIAM can currently resolve images 6500 times smaller than conventional light microscopes.