SCAPE Imaging Geometry and Neuronal Firing in Apical Dendrites in Mouse Brain (IMAGE)
Caption
This schematic depicts SCAPE's imaging geometry. The sample is illuminated by a thin sheet of light (blue), incident at an oblique angle. SCAPE achieves high speed imaging by sweeping this light sheet back and forth within the sample, achieved using a scanning mirror configured similarly to confocal microscopy. This optically sectioned plane is imaged onto a high speed sCMOS camera via the same objective lens. Unique de-scanning and image rotation optics ensure that the illuminated plane is always co-aligned with the camera plane, throughout its scan position. The end result is data equivalent to conventional light-sheet microscopy, but requiring a single, stationary objective lens, no sample translation, and consequently very high speed 3-D imaging. This unique configuration permits volumetric imaging of intact tissues including the awake, behaving mouse brain. While limited in penetration depth (since SCAPE is currently implemented with a 488 nm laser) spontaneous activity in apical dendrites in layers one and two of the mouse cortex can be resolved at >10 volumes per second. Panels show dendrites rendered from SCAPE data acquired in an awake behaving mouse with layer five neurons labeled with GCaMP5g. Renderings show dendritic branches corresponding to the colored time-courses shown below. Temporal resolution and signal to noise are sufficient to discern different properties of onset and decay dynamics within individual dendritic branches for single events.
Credit
Elizabeth Hillman, Columbia Engineering
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