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Introduction to Photomicrography

The use of photography to capture images in a microscope dates back to the invention of the photographic process. Early photomicrographs were remarkable for their quality, but the techniques were laborious and burdened with long exposures and a difficult process for developing emulsion plates. The primary medium for photomicrography was film until the past decade when improvements in electronic camera and computer technology made digital imaging cheaper and easier to use than conventional photography. This section will address photomicrography both on film and with electronic analog and digital imaging systems.

The quality of a photomicrograph, either digital or recorded on film, is dependent upon the quality of the microscopy. Film is a stern judge of how good the microscopy has been prior to capturing the image. It is essential that the microscope be configured using Köhler illumination, and that the field and condenser diaphragms are adjusted correctly and the condenser height is optimized. When properly adjusted, the microscope will yield images that have even illumination over the entire field of view and display the best compromise of contrast and resolution.

Almost all microscopists will, at some point, have the need or desire to record the images seen through the microscope. The main mechanism, for many years, of producing such photomicrographs was through the use of film, although in recent years most scientists have begun to capture images by means of electronic cameras. The main purpose of this tutorial is to enable the microscopist to record the observed images on film or digital media, and to do so with accuracy of image reproduction and with fidelity of color when color film is being used. The further aim is to empower the photomicrographer to secure excellent pictures without having to struggle through the already existing, far more complex reference literature. Use the links below to navigate to various topics in our discussions of photomicrography.

Troubleshooting Problems in Photomicrography - Photography through the microscope is undergoing a transition from film to digital imaging. New digital technologies are producing higher resolution micrographs, but the quality still falls short of that obtainable with film. Microscope configuration errors represent the greatest obstacle to quality photomicrographs, followed by errors in filter selection, film choice, aberration, dirt and debris, and processing mistakes.

Film Basics - There are a number of films made by manufacturers around the world that are suitable for photomicrography. This section discusses the properties, use, and technology of common photographic film.

Fundamentals of Film Exposure - Exposure of film, both black & white and color, is dependent upon the amount of time the film is exposed to light and the intensity of that light. Topics covered in this section include the reciprocity law, characteristic curves, exposure bracketing, exposure calculations, and filter factors.

Filters for Black & White Photomicrography - In black & white photography through the microscope, filters are used primarily to control contrast in the final image captured on film. They are also useful for overcoming optical aberration and increasing resolution by restriction of illuminating wavelengths.

Spectral Characteristics of Common Biological Stains - Compare the visible light absorption spectral data for common biological stains to determine suitability for use in black & white photomicrography.

Kodak Wratten Filters for Black & White Photomicrography - Visible light transmission regions for Kodak Wratten color filters are presented to aid microscopists in determining filter suitability for contrast and resolution control in black & white photomicrography.

Color Temperature - Based on the relationship between the temperature of a black body radiator and the energy distribution of its emitted light, this concept is of paramount importance in color photomicrography.

Filters for Color Photomicrography - A wide spectrum of filters are available to assist the microscopist in achieving the best quality images. These include color compensating filters, neutral density filters, didymium filters, filters to block ultraviolet light, and heat-absorbing filters.

Color Compensating Filters: Specifications and Spectral Data - Kodak color compensating filters are frequently used to fine-tune the color balance of tungsten-halogen microscope light sources for photomicrography with color films. This page is an index to our data on color compensating filter specifications, including spectral data for each individual filter set.

Film Cameras for Photomicrography - In photomicrography, regular camera lenses are not used because the microscope optical train, from the light source to the photoeyepiece, constitutes the image-forming lens assembly and illumination system. This section discusses various aspects of attaching a camera to the microscope and reviews automatic exposure cameras designed specifically for photomicrographic applications.

  • Olympus PM-30 Automatic Camera System - The Olympus PM-30 automatic camera system described in this section can easily handle the usual modes of microscopy including brightfield, phase contrast, differential interference contrast, polarized light, Hoffman modulation contrast, and darkfield.

Creative Photomicrography - By employing multiple exposure photomicrography, we have succeeded in generating a series of unusual micrographs which we have termed microscapes. These micrographs are intended to resemble surrealistic/alien landscapes.

Interactive Java and Flash Tutorials - Visit our gallery of interactive Java and Flash tutorials for user-controlled demonstrations about complex topics in photography through the microscope. Included are tutorials covering filters, color temperature, color balance and correction, aberrations, film exposure, and concepts in digital imaging technology.

Contributing Authors

Mortimer Abramowitz - Olympus America, Inc., Two Corporate Center Drive., Melville, New York, 11747.

Kenneth R. Spring - Scientific Consultant, Lusby, Maryland, 20657.

Brian O. Flynn, John C. Long, Kirill I. Tchourioukanov, and Michael W. Davidson - National High Magnetic Field Laboratory, 1800 East Paul Dirac Dr., The Florida State University, Tallahassee, Florida, 32310.


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