Pin-hole array correlation imaging: highly parallel fluorescence correlation spectroscopy

Biophys J. 2009 Jun 17;96(12):5050-9. doi: 10.1016/j.bpj.2009.03.023.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Colloids
  • Phycoerythrin / ultrastructure
  • Spectrometry, Fluorescence / instrumentation*
  • Spectrometry, Fluorescence / methods

Substances

  • Colloids
  • Phycoerythrin