TRANSCENDING THE APPARENT LIMITATIONS OF LASER SCANNING OPTICAL MICROSCOPY THROUGH CREATIVITY: FLUORESCENT LABELS, DETECTION SYSTEMS AND INSTRUMENTATION

Karl Garsha

Specialist in Light Microscopy, Imaging Technology Group, Beckman Institute, UIUC

1/31/2002

3rd Floor tower, 3269 Beckman 

Although laser scanning microscopy has only relatively recently become a practical solution for many researchers, the theory and technology behind these instruments represents the fruits of several decades of multi-disciplinary research.  Confocal and multiphoton laser scanning microscopes showcase the synergistic application of contributions from many fields including microscopy, video technology, electronics, computer science, laser technology, and optics.  Laser scanning confocal and multiphoton microscopies have become exceedingly popular modalities of imaging for a variety of disciplines including biological research, chemical analysis and materials testing.  Confocal and, more recently, multiphoton microscopy have gained acceptance as important imaging technologies owing to their versatility with regard to specimen preparation and the impressive capability to obtain images free from out of focus information.  The popularity and power of the laser scanning approach to microscopic imaging has encouraged researchers to further augment the versatility of these instruments by exploiting advancements from a variety of fields.  Existing and emerging approaches to super-fast laser illumination, clever fluorochrome design and combination, time-resolved signal detection, directed evolution of fluorescent protein markers, and sophisticated computational post-processing allows present day investigators to seek answers to questions that are ostensibly beyond the resolution and/or detection limits of the individual hardware components that make up a laser scanning microscope.  Knowledge of these approaches may serve to enhance investigators' individual research programs as well as accelerate the development and implementation of new and emerging methods of laser scanning microscopy.