Improving signal levels in intravital multiphoton microscopy using an objective correction collar Export

@article{citeulike:2862657, abstract = {Multiphoton microscopy has enabled biologists to collect high-resolution images hundreds of microns into biological tissues, including tissues of living animals. While the depth of imaging exceeds that possible from any other form of light microscopy, multiphoton microscopy is nonetheless generally limited to depths of less than a millimeter. Many of the advantages of multiphoton microscopy for deep tissue imaging accrue from the unique nature of multiphoton fluorescence excitation. However, the quadratic relationship between illumination level and fluorescence excitation makes multiphoton microscopy especially susceptible to factors that degrade the illumination focus. Here we examine the effect of spherical aberration on multiphoton microscopy in fixed kidney tissues and in the kidneys of living animals. We find that spherical aberration, as evaluated from axial asymmetry in the point-spread function, can be corrected by adjustment of the correction collar of a water immersion objective lens. Introducing a compensatory positive spherical aberration into the imaging system decreases the depth-dependence of signal levels in images collected from living animals, increasing signal by up to 50\%.}, author = {Muriello, Pamela A. and Dunn, Kenneth W.}, booktitle = {Optics in Life Sciences}, citeulike-article-id = {2862657}, citeulike-linkout-0 = {http://dx.doi.org/10.1016/j.optcom.2007.05.070}, citeulike-linkout-1 = {http://www.sciencedirect.com/science/article/B6TVF-4R34CP4-1/1/bd46e42e0f69fc93127ab491c44aa05b}, day = {1}, doi = {10.1016/j.optcom.2007.05.070}, journal = {Optics Communications}, keywords = {2pe, correction\_collar, method, psf, spherical\_abberation}, month = {April}, number = {7}, pages = {1806--1812}, posted-at = {2008-06-04 21:55:50}, priority = {2}, title = {Improving signal levels in intravital multiphoton microscopy using an objective correction collar}, url = {http://dx.doi.org/10.1016/j.optcom.2007.05.070}, volume = {281}, year = {2008} }

TY - JOUR ID - citeulike:2862657 L3 - citeulike-article-id:2862657 N2 - Multiphoton microscopy has enabled biologists to collect high-resolution images hundreds of microns into biological tissues, including tissues of living animals. While the depth of imaging exceeds that possible from any other form of light microscopy, multiphoton microscopy is nonetheless generally limited to depths of less than a millimeter. Many of the advantages of multiphoton microscopy for deep tissue imaging accrue from the unique nature of multiphoton fluorescence excitation. However, the quadratic relationship between illumination level and fluorescence excitation makes multiphoton microscopy especially susceptible to factors that degrade the illumination focus. Here we examine the effect of spherical aberration on multiphoton microscopy in fixed kidney tissues and in the kidneys of living animals. We find that spherical aberration, as evaluated from axial asymmetry in the point-spread function, can be corrected by adjustment of the correction collar of a water immersion objective lens. Introducing a compensatory positive spherical aberration into the imaging system decreases the depth-dependence of signal levels in images collected from living animals, increasing signal by up to 50%. IS - 7 JF - Optics Communications EP - 1812 TI - Improving signal levels in intravital multiphoton microscopy using an objective correction collar VL - 281 T2 - Optics in Life Sciences SP - 1806 KW - 2pe KW - correction_collar KW - method KW - psf KW - spherical_abberation AU - Muriello, Pamela AU - Dunn, Kenneth PY - 2008/04/01/ UR - http://dx.doi.org/10.1016/j.optcom.2007.05.070 ER -