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Automated microscopy system for mosaic acquisition and processing
Author(s) -
CHOW S. K.,
HAKOZAKI H.,
PRICE D. L.,
MACLEAN N. A. B.,
DEERINCK T. J.,
BOUWER J. C.,
MARTONE M. E.,
PELTIER S. T.,
ELLISMAN M. H.
Publication year - 2006
Publication title -
journal of microscopy
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.569
H-Index - 111
eISSN - 1365-2818
pISSN - 0022-2720
DOI - 10.1111/j.1365-2818.2006.01577.x
Subject(s) - image stitching , computer science , computer vision , software , mosaic , artificial intelligence , image processing , microscope , data acquisition , biological specimen , profiling (computer programming) , context (archaeology) , microscope image processing , digital image processing , computer graphics (images) , optics , image (mathematics) , biology , physics , paleontology , archaeology , history , programming language , operating system
Summary An automatic mosaic acquisition and processing system for a multiphoton microscope is described for imaging large expanses of biological specimens at or near the resolution limit of light microscopy. In a mosaic, a larger image is created from a series of smaller images individually acquired systematically across a specimen. Mosaics allow wide‐field views of biological specimens to be acquired without sacrificing resolution, providing detailed views of biological specimens within context. The system is composed of a fast‐scanning, multiphoton, confocal microscope fitted with a motorized, high‐precision stage and custom‐developed software programs for automatic image acquisition, image normalization, image alignment and stitching. Our current capabilities allow us to acquire data sets comprised of thousands to tens of thousands of individual images per mosaic. The large number of individual images involved in creating a single mosaic necessitated software development to automate both the mosaic acquisition and processing steps. In this report, we describe the methods and challenges involved in the routine creation of very large scale mosaics from brain tissue labelled with multiple fluorescent probes.