Abstract

In confocal fluorescence correlation microscopy (FCM) it is important to ensure that the correlation measurement is actually performed at the chosen location of the three-dimensional image of the specimen. We present a confocal FCM design that provides an automatic real-time readout of the location in the confocal microscopic image, which is aligned with the detector of the fluorescence correlation spectrometer. The design accomplishes this without using any special positioning device. The design is based on an apertured fluorescence detector placed close to the back aperture of the objective lens and can be easily incorporated into virtually any confocal microscope. We demonstrate the method by performing FCM measurements of a dye diffusing on a cell membrane.

© 2005 Optical Society of America

Low-cost, scalable laser scanning module for real-time reflectance and fluorescence confocal microscopy

Derrick R. Chou, Bradley A. Bower, and Adam Wax
Appl. Opt. 44(11) 2013-2018 (2005)

EMCCD-based spectrally resolved fluorescence correlation spectroscopy

Felix Bestvater, Zahir Seghiri, Moon Sik Kang, Nadine Gröner, Ji Young Lee, Kang-Bin Im, and Malte Wachsmuth
Opt. Express 18(23) 23818-23828 (2010)

Fluorescence correlation spectroscopy: inception, biophysical experimentations, and prospectus

Watt W. Webb
Appl. Opt. 40(24) 3969-3983 (2001)

Electron multiplying CCD based detection for spatially resolved fluorescence correlation spectroscopy

Markus Burkhardt and Petra Schwille
Opt. Express 14(12) 5013-5020 (2006)

Dual-Color Fluorescence Cross-Correlation Spectroscopy on a Single Plane Illumination Microscope (SPIM-FCCS)

Jan Wolfgang Krieger, Anand Pratap Singh, Christoph S. Garbe, Thorsten Wohland, and Jörg Langowski
Opt. Express 22(3) 2358-2375 (2014)