Light emitting diodes: Light engines to simplify and economize advanced microscopy

Abstract

THE old adage, ‘‘A picture is worth a thousand words,’’ is especially true in biological research, with microscopic images serving to mark the development of key concepts in biology since Hooke first described ‘‘cells’’ in thin slices of cork (1). More than 400 years later, microscopic imaging still serves as a primary data source for biological research, with increasingly advanced techniques that provide information on molecular scale interactions and dynamics. Such advanced molecular imaging techniques take advantage of ever more sophisticated light sources, optics, detectors, and image processing methods, many of which can be quite complex and expensive. At the same time, new technologies and production methods, many driven by the consumer market, have resulted in key components of imaging systems becoming smaller, simpler, and cheaper. The convergence of these effects is having profound effects on our options for light sources for microscopic imaging. The properties of the illumination source are critical to many advanced optical microscopy methods. Lasers play a key role in many of these owing to the monochromatic and coherent nature of their light, plus their ability to be pulsed and otherwise controlled. However, illumination with high-pressure mercury gas discharge or xenon lamps is still the standard for most imaging applications. These light sources provide bright, broad-spectrum excitation (300–900 nm) that can be easily selected by excitation filters for fluorescent imaging applications. However, there are quite a few disadvantages that limit the wide-spread of mercury or xenon lamps-based microscopy system in biological applications. First, sometimes these traditional light sources are too bright and harmful for the living organisms being studied. Excessive brightness can cause rapid photo-bleaching of the dyes used to stain the cells/ tissues. UV light especially, if not blocked, can have significant deleterious effects on live cells. Second, there are substantial costs involved in using mercury or xenon lamps because of their relatively short lifespan. The life of a mercury lamp is usually several hundred hours, and the intensity of these lamps decays progressively during this time. In addition, gas discharge lamps require some time to warm up before lamp intensity reaches steady state. Therefore, once mercury lamps are turned on, they have to be left on for hours to enable fluorescent measurements as needed without delay, thereby further shortening the lamp lifetime. Third, there are also some hazards issues of using mercury or xenon lamps. These lamps generate a significant amount of heat and therefore introduce complications when used in a confined space. When mercury lamps die, they can explode and damage the lenses or mirrors within the lamp housing. These factors combine to make gas discharge lamps inconvenient for lab-based applications and impractical for field applications, including use in portable, miniature, or low cost imaging devices. By contrast, light-emitting diodes (LEDs) show great pro