Modern Laser Scanning Confocal Microscopy.

Since its commercialization in the late 1980's, confocal laser scanning microscopy (CLSM) has since become one of the most prevalent fluorescence microscopy techniques for three-dimensional structural studies of biological cells and tissues. The flexibility of the approach has enabled its application in a diverse array of studies, from the fast imaging of dynamic processes in living cells, to meticulous morphological analyses of tissues, and co-localization of protein expression patterns. In this chapter, we introduce the principles of confocal microscopy and discuss how the approach has become a mainstay in the biological sciences.

Nonlinear optical imaging of extracellular matrix proteins.

Over the last 2 decades, nonlinear imaging methods such as multiharmonic imaging microscopy (MHIM) have become powerful approaches for the label-free visualization of biological structures. Multiharmonic signals are generated when an intense electromagnetic field propagates through a sample that either has a specific molecular orientation or exhibits certain physical properties. It can provide complementary morphological information when integrated with other nonlinear optical imaging techniques such as two-photon excitation (TPE).

Laminar circuit formation in the vertebrate retina.

Neuronal function depends on the accurate wiring between pre- and postsynaptic cells. Determining the mechanisms underlying precision in neuronal connectivity is challenging because of the complexity of the nervous system. In diverse parts of the nervous system, regions of synaptic contact are organized into distinct parallel layers, or laminae, that are correlated with distinct functions.