Pin-hole array correlation imaging: highly parallel fluorescence correlation spectroscopy.
In this work, we describe pin-hole array correlation imaging, a multipoint version of fluorescence correlation spectroscopy, based upon a stationary Nipkow disk and a high-speed electron multiplying charged coupled detector. We characterize the system and test its performance on a variety of samples, including 40 nm colloids, a fluorescent protein complex, a membrane dye, and a fluorescence fusion protein. Our results demonstrate that pin-hole array correlation imaging is capable of simultaneously performing tens or hundreds of fluorescence correlation spectroscopy-style measurements in cells, with sufficient sensitivity and temporal resolution to study the behaviors of membrane-bound and soluble molecules labeled with conventional chemical dyes or fluorescent proteins.
Needleman
Daniel J
DJ
Department of Systems Biology, Harvard Medical School, Boston, Massachusetts, USA. daniel_needleman@hms.harvard.edu
Xu
Yangqing
Y
Mitchison
Timothy J
TJ
D003102:Colloids; D010799:Phycoerythrin / Q000648:ultrastructure; D013050:Spectrometry, Fluorescence / Q000295:instrumentation* / Q000379:methods
D003102:Colloids; D010799:Phycoerythrin