HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) Supplemental File All Versions of this Article: 293/3/C1073    most recent 00586.2006v1 Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in ISI Web of Science Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via ISI Web of Science (3) Citing Articles via Google Scholar Google Scholar Articles by Picht, E. Articles by Bers, D. M. Search for Related Content PubMed PubMed Citation Articles by Picht, E. Articles by Bers, D. M.

METHODS IN CELL PHYSIOLOGY

SparkMaster: automated calcium spark analysis with ImageJ

Department of Physiology, Stritch School of Medicine, Loyola University Chicago, Maywood, Illinois

Submitted 21 November 2006 ; accepted in final form 20 March 2007

Ca sparks are elementary Ca-release events from intracellular Ca stores that are observed in virtually all types of muscle. Typically, Ca sparks are measured in the line-scan mode with confocal laser-scanning microscopes, yielding two-dimensional images (distance vs. time). The manual analysis of these images is time consuming and prone to errors as well as investigator bias. Therefore, we developed SparkMaster, an automated analysis program that allows rapid and reliable spark analysis. The underlying analysis algorithm is adapted from the threshold-based standard method of spark analysis developed by Cheng et al. (Biophys J 76: 606–617, 1999) and is implemented here in the freely available image-processing software ImageJ. SparkMaster offers a graphical user interface through which all analysis parameters and output options are selected. The analysis includes general image parameters (number of detected sparks, spark frequency) and individual spark parameters (amplitude, full width at half-maximum amplitude, full duration at half-maximum amplitude, full width, full duration, time to peak, maximum steepness of spark upstroke, time constant of spark decay). We validated the algorithm using images with synthetic sparks embedded into backgrounds with different signal-to-noise ratios to determine an analysis criteria at which a high sensitivity is combined with a low frequency of false-positive detections. Finally, we applied SparkMaster to analyze experimental data of sparks measured in intact and permeabilized ventricular cardiomyocytes, permeabilized mammalian skeletal muscle, and intact smooth muscle cells. We found that SparkMaster provides a reliable, easy to use, and fast way of analyzing Ca sparks in a wide variety of experimental conditions.

myocytes; sarcoplasmic reticulum; confocal microscopy

This article has been cited by other articles:

				J. D. Pickering, E. White, A. M. Duke, and D. S. Steele DHPR activation underlies SR Ca2+ release induced by osmotic stress in isolated rat skeletal muscle fibers J. Gen. Physiol., May 1, 2009; 133(5): 511 - 524. [Abstract] [Full Text] [PDF]

				T. Hayashi, M. E. Martone, Z. Yu, A. Thor, M. Doi, M. J. Holst, M. H. Ellisman, and M. Hoshijima Three-dimensional electron microscopy reveals new details of membrane systems for Ca2+ signaling in the heart J. Cell Sci., April 1, 2009; 122(7): 1005 - 1013. [Abstract] [Full Text] [PDF]

				H. Tateishi, M. Yano, M. Mochizuki, T. Suetomi, M. Ono, X. Xu, H. Uchinoumi, S. Okuda, T. Oda, S. Kobayashi, et al. Defective domain-domain interactions within the ryanodine receptor as a critical cause of diastolic Ca2+ leak in failing hearts Cardiovasc Res, February 15, 2009; 81(3): 536 - 545. [Abstract] [Full Text] [PDF]

				A. V. Zima, J. Qin, M. Fill, and L. A. Blatter Tricyclic antidepressant amitriptyline alters sarcoplasmic reticulum calcium handling in ventricular myocytes Am J Physiol Heart Circ Physiol, November 1, 2008; 295(5): H2008 - H2016. [Abstract] [Full Text] [PDF]

				A. V. Zima, E. Picht, D. M. Bers, and L. A. Blatter Termination of Cardiac Ca2+ Sparks: Role of Intra-SR [Ca2+], Release Flux, and Intra-SR Ca2+ Diffusion Circ. Res., October 10, 2008; 103(8): e105 - e115. [Abstract] [Full Text] [PDF]

				H. Cheng and W. J. Lederer Calcium Sparks Physiol Rev, October 1, 2008; 88(4): 1491 - 1545. [Abstract] [Full Text] [PDF]

				T. L. Domeier, A. V. Zima, J. T. Maxwell, S. Huke, G. A. Mignery, and L. A. Blatter IP3 receptor-dependent Ca2+ release modulates excitation-contraction coupling in rabbit ventricular myocytes Am J Physiol Heart Circ Physiol, February 1, 2008; 294(2): H596 - H604. [Abstract] [Full Text] [PDF]