In this work, we combined the advantages of second-harmonic generation (SHG) with a random access (RA) excitation scheme to realize a new microscope (RA-SHG) capable of optically recording fast membrane potential events occurring in a wide-field configuration. The RA-SHG microscope in combination with a bulk staining method with FM4-64 was used to simultaneously record electrical activity from clusters of Purkinje cells (PCs) in acute cerebellar slices. Spontaneous electrical activity was also monitored simultaneously in pairs of neurons, where APs were recorded in a single trial without averaging. These results show the strength of this technique to describe the temporal dynamics of neuronal assemblies.
Action potential detection by non-linear microscopy / L. Sacconi; J. Lotti; R. P. O'Connor; J. Mapelli; D. Gandolfi; E. D'Angelo; F. S. Pavone. - STAMPA. - (2009), pp. 71613F-716113F. (Intervento presentato al convegno Progress in Biomedical Optics and Imaging).
Action potential detection by non-linear microscopy
SACCONI, LEONARDO;LOTTI, JACOPO;PAVONE, FRANCESCO SAVERIO
2009
Abstract
In this work, we combined the advantages of second-harmonic generation (SHG) with a random access (RA) excitation scheme to realize a new microscope (RA-SHG) capable of optically recording fast membrane potential events occurring in a wide-field configuration. The RA-SHG microscope in combination with a bulk staining method with FM4-64 was used to simultaneously record electrical activity from clusters of Purkinje cells (PCs) in acute cerebellar slices. Spontaneous electrical activity was also monitored simultaneously in pairs of neurons, where APs were recorded in a single trial without averaging. These results show the strength of this technique to describe the temporal dynamics of neuronal assemblies.
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