Supriya Balaji Ramachandran1, Kevin D Gillis2. 1. Department of Bioengineering, 254 Agricultural Engineering, Columbia, MO, USA; Dalton Cardiovascular Research Center, University of Missouri, Columbia, MO, USA. 2. Department of Bioengineering, 254 Agricultural Engineering, Columbia, MO, USA; Department of Medical Pharmacology and Physiology, 1 Hospital Dr., MO, USA; Dalton Cardiovascular Research Center, University of Missouri, Columbia, MO, USA. Electronic address: GillisK@missouri.edu.
Abstract
BACKGROUND: Quantal exocytosis of oxidizable neurotransmitters can be detected as spikes of amperometric current using electrochemical microelectrodes. Measurements of spike parameters indicate the maximal transmitter flux, flux duration, and amount of transmitter released from individual vesicles. Automated analysis algorithms need to reject spikes that overlap in time. In addition, many spikes are preceded by small amplitude "foot" signals, attributed to slow release of transmitter through a fusion pore. Accurate pre-spike baseline determination is essential for estimating fusion-pore duration and the amount of transmitter released through the fusion pore. NEW METHOD: We developed an estimation approach that is based on fitting a multi-exponential function to the data. Our previously described matched-filter algorithm is used to identify the sections of data to fit and provides seed values to facilitate convergence of the iterative fit. The new estimation algorithm includes overlap rejection, a two-step fitting procedure and a novel baseline estimation procedure. RESULTS: Histograms of spike parameters demonstrate excellent agreement of the new approach with manually computed parameters. COMPARISON WITH EXISTING METHODS: Parameter estimates generated using the new approach are closer to blind manual estimates than commonly used existing methods. The improved performance is due to better detection of valid spikes and rejection of overlapping spikes. Moreover, since the complete time course of the spike is fit to a function, more complete information about the spike time course is captured. CONCLUSIONS: The matched-filter seeded algorithm reliably rejects overlaps and estimates spike and foot signal parameters in a fully automated manner.
BACKGROUND: Quantal exocytosis of oxidizable neurotransmitters can be detected as spikes of amperometric current using electrochemical microelectrodes. Measurements of spike parameters indicate the maximal transmitter flux, flux duration, and amount of transmitter released from individual vesicles. Automated analysis algorithms need to reject spikes that overlap in time. In addition, many spikes are preceded by small amplitude "foot" signals, attributed to slow release of transmitter through a fusion pore. Accurate pre-spike baseline determination is essential for estimating fusion-pore duration and the amount of transmitter released through the fusion pore. NEW METHOD: We developed an estimation approach that is based on fitting a multi-exponential function to the data. Our previously described matched-filter algorithm is used to identify the sections of data to fit and provides seed values to facilitate convergence of the iterative fit. The new estimation algorithm includes overlap rejection, a two-step fitting procedure and a novel baseline estimation procedure. RESULTS: Histograms of spike parameters demonstrate excellent agreement of the new approach with manually computed parameters. COMPARISON WITH EXISTING METHODS: Parameter estimates generated using the new approach are closer to blind manual estimates than commonly used existing methods. The improved performance is due to better detection of valid spikes and rejection of overlapping spikes. Moreover, since the complete time course of the spike is fit to a function, more complete information about the spike time course is captured. CONCLUSIONS: The matched-filter seeded algorithm reliably rejects overlaps and estimates spike and foot signal parameters in a fully automated manner.
Authors: R M Wightman; J A Jankowski; R T Kennedy; K T Kawagoe; T J Schroeder; D J Leszczyszyn; J A Near; E J Diliberto; O H Viveros Journal: Proc Natl Acad Sci U S A Date: 1991-12-01 Impact factor: 11.205
Authors: Yan Yang; Tim J Craig; Xiaohui Chen; Leonora F Ciufo; Masami Takahashi; Alan Morgan; Kevin D Gillis Journal: J Gen Physiol Date: 2007-03 Impact factor: 4.086