Spatiotemporal Rank Filtering Improves Image Quality Compared to Frame Averaging in 2-Photon Laser Scanning Microscopy

PLoS One

Kaitlin Corbin¹  Matthew F. Krummel²  Henry Pinkard³ 

[1] Biological Imaging Development Center, University of California San Francisco, San Francisco, California, United States of America;Computational Biology Graduate Group, University of California Berkeley, Berkeley, California, United States of America;Department of Pathology, University of California San Francisco, San Francisco, California, United States of America

关键词: Signal filtering;    Background signal noise;    Fluorescence imaging;    Photons;    Lymph nodes;    Imaging techniques;    In vivo imaging;    Fluorescence;   

DOI  :  10.1371/journal.pone.0150430

学科分类：医学（综合）

来源: Public Library of Science

【 摘 要 】

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.

【 授权许可】

CC BY