Target-locking acquisition with real-time confocal (TARC) microscopy

We present a real-time target-locking confocal microscope that follows an object moving along an arbitrary path, even as it simultaneously changes its shape, size and orientation. This Target-locking Acquisition with Realtime Confocal (TARC) microscopy system integrates fast image processing and rap...

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Bibliographic Details
Published in:Optics express 2007-07, Vol.15 (14), p.8702-8712
Main Authors: Lu, Peter J, Sims, Peter A, Oki, Hidekazu, Macarthur, James B, Weitz, David A
Format: Article
Language:English
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Summary:We present a real-time target-locking confocal microscope that follows an object moving along an arbitrary path, even as it simultaneously changes its shape, size and orientation. This Target-locking Acquisition with Realtime Confocal (TARC) microscopy system integrates fast image processing and rapid image acquisition using a Nipkow spinning-disk confocal microscope. The system acquires a 3D stack of images, performs a full structural analysis to locate a feature of interest, moves the sample in response, and then collects the next 3D image stack. In this way, data collection is dynamically adjusted to keep a moving object centered in the field of view. We demonstrate the system's capabilities by target-locking freely-diffusing clusters of attractive colloidal particles, and activelytransported quantum dots (QDs) endocytosed into live cells free to move in three dimensions, for several hours. During this time, both the colloidal clusters and live cells move distances several times the length of the imaging volume.
ISSN:1094-4087
1094-4087
DOI:10.1364/OE.15.008702