Scattering suppression and confocal detection in multifocal multiphoton microscopy

Martini J, Andresen V, Anselmetti D. Scattering suppression and confocal detection in multifocal multiphoton microscopy. JOURNAL OF BIOMEDICAL OPTICS. 2007;12(3): 034010.We have developed a new descanned parallel (32-fold) pinhole and photomultiplier detection array for multifocal multiphoton microscopy that effectively reduces the blurring effect originating from scattered fluorescence photons in strongly scattering biological media. With this method, we achieve a fourfold improvement in photon statistics for detecting ballistic photons and an increase in spatial resolution by 21% in the lateral and 35% in the axial direction compared to single-beam non-descanned multiphoton microscopy. The new detection concept has been applied to plant leaves and pollen grains to verify the improvements in imaging quality. (C) 2007 Society of Photo-Optical Instrumentation Engineers