Recent advances in optical microscopy methods for subcellular imaging of thick biological tissues.

Abstract

Optical microscopy has been widely applied in cellular and subcellular imaging. Conventional light microscopes, however, have rather limited imaging depth and are limited to imaging only mechanically sectioned thin samples. Multiphoton microscopy and optical coherence microscopy are common techniques for diffraction-limited imaging beyond an imaging depth of 0.5 mm. Focal modulation microscopy is a novel method that combines confocal spatial filtering with focal modulation to reject out-of-focus backgrounds. Focal modulation microscopy has demonstrated an imaging depth comparable to those of multiphoton microscopy and optical coherence microscopy, near-real-time image acquisition, and capability with a multiple contrast mechanism.