FEMTO3D-ACOUSTOOPTIC

MEASUREMENT OF PROPAGATION SPEED IN MULTIPLE DENDRITES SIMULTANEOUSLY


Four dendritic segments were simultaneously measured in 3D. Dots in the diagramm of the cells represents the points of measurement. 3D Ca2+ responses recorded during a SPW-EPSPs are represented on the right side of the image.


Following signal propagation and integration across a neuron and recording the concerted activity of hundreds of neurons pose distinct challenges, and the design of the Femto3D-AcoustoOptic imaging system has been focused on tackling both of these problems.




        


EXAMPLE OF MULTIPLE SITE DENDRITIC IMAGING



                                                                                                       

3D AO scanning (top, z stack): a 3D view of the dendritic arbor of a CA1 pyramidal cell imaged; spheres represent the measurement locations. Maximum intensity z-projection image of the same neuron; recorded dendrites are numbered. Schema of the apical trunk and the dendritic branches of the neuron is showing calcium transients recorded near-simultaneously in each of the dendrites, averaged from five traces. Repetition rate of the 3D point scanning was 80 Hz. Dendritic Ca2+ transients measured from the same dendritic point of the apical trunk (average of five traces) and corresponding somatic voltage traces (Vm).



EXAMPLE FOR 3D-AO TWO-PHOTON IN VIVO MEASUREMENTS 



Three dimensional random-access measurement of network activity in mouse V1. (a) In vivo experimental arrangement (sketch). Visual stimulation was induced by continuously moving bars (in 45-degree steps at eight directions of motion). (b) Maximal intensity side- and z-projection of the z-stack. 375 autodetected neuronal locations. (c) Spheres: from the same 375 neuronal location 3D Ca2+ responses was measured (visual stimulation: moving bar at +45°). Rows: single cells from a single 3D measurement. (d) Ca2+ transients examples from responding neurons in c. (e) Stimulation with 90° oriented stimulus  (at +135°) induced smaller responses from the same neurons with a different activation pattern.


See also:
Gergely Katona, Gergely Szalay, Pál Maák, Attila Kaszás, Máté Veress, Dániel Hillier, Balázs Chiovini, E Sylvester Vizi, Botond Roska & Balázs Rózsa
Fast two-photon in vivo imaging with three-dimensional random-access scanning in large tissue volumes,
Nature Methods (2012)

MEASUREMENT OF PROPAGATING CA2+ WAVES

      
     

Left panel shows the 3D Ca2+ responses along a single dendrite during a sharpe-wave event. Note that the high scanning speed makes the propagation 

clearly distingusihable. The left panel shows the half maximum of the local transients versus distance from the soma. Propagation speed can be 

determined by linear fit.

 
See also:
B Chiovini, G F Turi, G Katona, A Kaszas, D Palfi, P Maak, G Szalay, M F Szabo, Z Szadai, Sz Kali and B Rozsa
Dendritic spikes induce ripples in parvalbumin interneurons during hippocampal sharp waves. 
Neuron (2014)

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