Data from: Spatiotemporal rank filtering improves image quality compared to frame averaging in 2-photon laser scanning microscopy
Data files
May 05, 2016 version files 492.24 MB
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Beads + masks.zip
46.78 MB
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VPD 450 T cell 30 frames 82 Zslices.tif
413.96 MB
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XCR1Venus.tif
31.51 MB
Abstract
Live imaging of biological specimens using optical microscopy is limited by tradeoffs between spatial and temporal resolution, depth into intact samples, and phototoxicity. Two-photon laser scanning microscopy (2P-LSM), the gold standard for imaging turbid samples in vivo, has conventionally constructed images with sufficient signal-to-noise ratio (SNR) generated by sequential raster scans of the focal plane and temporal integration of the collected signals. Here, we describe spatiotemporal rank filtering, a nonlinear alternative to temporal integration, which makes more efficient use of collected photons by selectively reducing noise in 2P-LSM images during acquisition. This results in much higher SNR while preserving image edges and fine details. Practically, this allows for at least a four fold decrease in collection times, a substantial improvement for time-course imaging in biological systems.