Acousto-optical Scanning–Based High- Speed 3D Two-Photon Imaging in Vivo
  • Metaadatok
Tartalom: http://real.mtak.hu/65065/
Archívum: REAL
Gyűjtemény: Status = Published
Subject = Q Science / természettudomány: QC Physics / fizika
Subject = Q Science / természettudomány: QC Physics / fizika: QC02 Optics / fénytan
Type = Book Section
Cím:
Acousto-optical Scanning–Based High- Speed 3D Two-Photon Imaging in Vivo
Létrehozó:
Rózsa, Balázs
Gergely, Szalay
Gergely, Katona
Kiadó:
Springer
Dátum:
2016-01-15
Téma:
QC Physics / fizika
QC02 Optics / fénytan
Tartalmi leírás:
Recording of the concerted activity of neuronal assemblies and the dendritic and axonal signal integration of downstream neurons pose different challenges, preferably a single recording system should perform both operations. We present a three-dimensional (3D), high-resolution, fast, acousto-optic two-photon microscope with random-access and continuous trajectory scanning modes reaching a cubic millimeter scan range (now over 950 × 950 × 3000 μm3) which can be adapted to imaging different spatial scales. The resolution of the system allows simultaneous functional measurements in many fine neuronal processes, even in dendritic spines within a central core (>290 × 290 × 200 μm3) of the total scanned volume. Furthermore, the PSF size remained sufficiently low (PSFx < 1.9 μm, PSFz < 7.9 μm) to target individual neuronal somata in the whole scanning volume for simultaneous measurement of activity from hundreds of cells. The system contains new design concepts: it allows the acoustic frequency chirps in the deflectors to be adjusted dynamically to compensate for astigmatism and optical errors; it physically separates the z-dimension focusing and lateral scanning functions to optimize the lateral AO scanning range; it involves a custom angular compensation unit to diminish off-axis angular dispersion introduced by the AO deflectors, and it uses a high-NA, wide-field objective and high-bandwidth custom AO deflectors with large apertures. We demonstrate the use of the microscope at different spatial scales by first showing 3D optical recordings of action potential back propagation and dendritic Ca2+ spike forward propagation in long dendritic segments in vitro, at near-microsecond temporal resolution. Second, using the same microscope we show volumetric random-access Ca2+ imaging of spontaneous and visual stimulation-evoked activity from hundreds of cortical neurons in the visual cortex in vivo. The selection of active neurons in a volume that respond to a given stimulus was aided by the real-time data analysis and the 3D interactive visualization accelerated selection of regions of interest.
Nyelv:
magyar
Típus:
Book Section
PeerReviewed
Formátum:
text
Azonosító:
Rózsa, Balázs and Gergely, Szalay and Gergely, Katona (2016) Acousto-optical Scanning–Based High- Speed 3D Two-Photon Imaging in Vivo. In: Advanced Patch-Clamp Analysis for Neuroscientists. Neuromethods . Springer, pp. 51-61. ISBN 978-1-4939-3411-9
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