Removing bleaching artifacts from voltage sensitive dye recordings with ICA

Experimentally, we recorded from Retzius cells of an isolated midbody ganglion of the electrophysiologically well- characterized leech nervous system. VSD imaging (Figure A) based on a new VSD dye with a very good signal-to-noise ratio [2] was combined with simultaneous intracellular recording of the membrane potential (Figure B) and electrical stimulation. VSD signals obtained from the region of the Retzius cell body (Figure 1A and 1C, red) and from a slightly larger region (Figure 1A and 1C, blue) were used as input signals to ICA. Of the two components returned by the algorithm one decreased exponentially, while the other resembled neuronal activity. Figure 1D shows an exponential fit of the component representing bleaching (green), and the estimated neuronal activity (red). Taking the lower sampling rate of the VSD signal (100 Hz vs 10,000 Hz) into account, this estimate reflects the dynamics of the intracellular recording (1B) well. While PCA was not able to separate bleaching from neuronal signals (not shown), the detrend method also yielded good results (Figure 1E). This method fits a piecewise linear line (1E, green) to the VSD signal, representing bleaching effect. However, the difference (1E, red) between the original trace and this line represents the graded de-and hyperpolarisations of the membrane potential seen in the intracellular recording (1B) less well than the ICA result (1D, red).