ABSTRACT

The opportunity for exploring basic fungal activity from the subcellular to the tissue level has never been greater. A multitude of technological advances in molecular techniques, fluorescent probe chemistry, and optical, computer, and laser technologies have contributed to an exponential growth in imaging for solving critical biological problems. Since its inception, confocal microscopy has proven itself to be an invaluable approach and a whole new way of visualizing biological cells and tissues. The ability to extract high-contrast, high-resolution noninvasive optical sections in both time and space has led to a transformation in how imaging is done in a variety of biological disciplines. Nearly 50 years after its invention, the technology continues to make rapid advancements that expand its utility beyond simple image acquisition. Hardware improvements have been made in all aspects of systems design, including acousto-optical tunable filters; solid-state, diode, and pulsed lasers; use of fiber optics; significant improvements in detection efficiency and the collection of spectral information, to name a few. Multiphoton microscopy has optical sectioning capabilities, much like confocal microscopy, and although these technologies share many attributes and capabilities, multiphoton extends the potential for viewing deeper into samples, is inherently less affected by highly scattering samples, and by virtue of nearinfrared (NIR) light source, can be less harmful to living organisms.