Combined analysis of intracellular calcium with dual excitation fluorescence photometry and imaging

Dietmar Uttenweiler; Reinhold Wojciechowski; Makoto Makabe; Claudia Veigel; Rainer H.A. Fink

doi:10.1117/12.210736

1 October 1995 Combined analysis of intracellular calcium with dual excitation fluorescence photometry and imaging

Dietmar Uttenweiler, Reinhold Wojciechowski, Makoto Makabe, Claudia Veigel, Rainer H.A. Fink

Author Affiliations +

Optical Engineering, Vol. 34, Issue 10, (October 1995). https://doi.org/10.1117/12.210736

Abstract

We have developed an integrated microscopy system combining fast dual-excitation fluorescence photometry and digital image analysis with high spatial resolution, based mainly on standard components. With the combination of these well-established techniques in one setup it is possible to monitor intracellular calcium with both sufficiently high temporal and high spatial resolution on the same preparation for many biological applications. Our system consists of a commercially available dual-excitation photometric system, an attached intensified charge coupled device (ICCD) camera, and a frame grabber board. With this integrated setup one can easily switch between the fast photometric mode (v_ratio = 100 Hz) and the imaging mode (v_ratio = 4.l7 up to 17 Hz). We used the system to record Fura-2 calcium images (340/380 nm ratios), which were correlated with the faster spot measurements and were analyzed by means of image processing. As an example for its application we reconstructed caffeine-induced calcium transients released from the sarcoplasmic reticulum of isolated and permeabilized skeletal muscle fiber preparations. Such a combined technique will also be important for cellular studies using other fluorescence indicators. Additionally, the described system has an external trigger facility that enables combination with other cell physiological methods, e.g., electrophysiological techniques.

Citation Download Citation

Dietmar Uttenweiler, Reinhold Wojciechowski, Makoto Makabe, Claudia Veigel, and Rainer H.A. Fink "Combined analysis of intracellular calcium with dual excitation fluorescence photometry and imaging," Optical Engineering 34(10), (1 October 1995). https://doi.org/10.1117/12.210736

Published: 1 October 1995

ACCESS THE FULL ARTICLE

INSTITUTIONAL
Select your institution to access the SPIE Digital Library.

SELECT YOUR INSTITUTION

PERSONAL
Sign in with your SPIE account to access your personal subscriptions or to use specific features such as save to my library, sign up for alerts, save searches, etc.

PERSONAL SIGN IN

No SPIE Account? Create one

PURCHASE THIS CONTENT

SUBSCRIBE TO DIGITAL LIBRARY

50 downloads per 1-year subscription

Members: $195

Non-members: $335 ADD TO CART

25 downloads per 1 - year subscription

Members: $145

Non-members: $250 ADD TO CART

PURCHASE SINGLE ARTICLE

Includes PDF, HTML & Video, when available