NDLI: Spatial calibration of structured illumination fluorescence microscopy using capillary tissue phantoms.

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Spatial calibration of structured illumination fluorescence microscopy using capillary tissue phantoms.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Lee, Grace S. Miele, Lino F. Turhan, Aslihan Lin, Miao Hanidziar, Dusan Konerding, Moritz A. Mentzer, Steven J.
Description	Country affiliation: United States Author Affiliation: Lee GS ( Laboratory of Adaptive and Regenerative Biology, Brigham and Women's Hospital, Department of Surgery, Harvard Medical School, Boston, Massachusetts 02115, USA.)
Abstract	Quantitative assessment of microvascular structure is relevant to the investigations of ischemic injury, reparative angiogenesis and tumor revascularization. In light microscopy applications, thick tissue specimens are necessary to characterize microvascular networks; however, thick tissue leads to image distortions due to out-of-focus light. Structured illumination confocal microscopy is an optical sectioning technique that improves contrast and resolution by using a grid pattern to identify the plane-of-focus within the specimen. Because structured illumination can be applied to wide-field (nonscanning) microscopes, the microcirculation can be studied by sequential intravital and confocal microscopy. To assess the application of structured illumination confocal microscopy to microvessel imaging, we studied cell-sized microspheres and fused silica microcapillary tissue phantoms. As expected, structured illumination produced highly accurate images in the lateral (X-Y) plane, but demonstrated a loss of resolution in the Z-Y plane. Because the magnitude of Z-axis distortion was variable in complex tissues, the silica microcapillaries were used as spatial calibration standards. Morphometric parameters, such as shape factor, were used to empirically optimize Z-axis software compression. We conclude that the silica microcapillaries provide a useful tissue phantom for in vitro studies as well as spatial calibration standard for in vivo morphometry of the microcirculation.
File Format	HTM / HTML
ISSN	1059910X
e-ISSN	10970029
DOI	10.1002/jemt.20647
Journal	Microscopy Research and Technique
Issue Number	2
Volume Number	72
Language	English
Publisher	Wiley-Liss
Publisher Date	2009-02-01
Publisher Place	United States
Access Restriction	Open
Subject Keyword	Discipline Chemistry Techniques Capillaries Ultrastructure Image Processing, Computer-assisted Microscopy, Confocal Microscopy, Fluorescence Microvessels Phantoms, Imaging Animals Calibration Carbocyanines Chemistry Data Interpretation, Statistical Instrumentation Mice Mice, Inbred C57bl Microcirculation Microspheres Nonlinear Dynamics Silicon Dioxide Research Support, N.i.h., Extramural
Content Type	Text
Resource Type	Article
Subject	Anatomy Histology Instrumentation Medical Laboratory Technology

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Spatial Calibration of Structured Illumination Fluorescence Microscopy using Capillary Tissue Phantoms

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