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We present endoscopic focal modulation microscopy (FMM) for minimally invasive imaging of deep tissue with high contrast, and then compare the results using endoscopic confocal microscopy (CM). Deep tissue imaging is achieved using a needle-like endoscopic probe based on gradient-index (GRIN) lenses. A tissue-like phantom with fluorescent micro-beads and rat kidney tubules were imaged through the endoscopic probe with FMM. FMM effectively rejected the background signals from the out-of-focus plane, thereby enhancing the image contrast and the optical sectioning ability. The combination of the GRIN endoscopic probe and FMM provides deep tissue imaging with better contrast than endoscopic CM. [ABSTRACT FROM AUTHOR]

Abstract: Image formation in focal modulation microscopy using quadrant apertures (QFMM) is presented. The spatial resolution is discussed: compared with confocal microscopy, QFMM can simultaneously enhance the axial and transverse resolution. In contrast to focal modulation microscopy using D-shaped apertures (DFMM), QFMM can maintain x–y symmetry of spatial resolution, thus causing less confusion in imaging interpretation. The capability of background rejection is investigated, showing QFMM has the potential to increase imaging penetration depth. The signal level is also analyzed, indicating that the preferred detector pinhole radius is greater than for DFMM, resulting in a higher signal level. [ABSTRACT FROM AUTHOR]

The historical development of multiphoton microscopy is described, starting with a review of two-photon absorption, and including two- and three-photon fluorescence microscopies, and second- and third-harmonic generation microscopies. The effects of pulse length on signal strength and breakdown are considered. Different contrast mechanisms, including use of nanoparticles, are discussed. Two new promising techniques that can be applied to multiphoton microscopy are described. [ABSTRACT FROM AUTHOR]