Time-Dependent Image Restoration of Low-SNR Live-Cell Ca2 Fluorescence Microscopy Data

International Journal of Molecular Sciences

Sukanya A. Kannabiran ¹  Björn-Philipp Diercks ¹  Valerie J. Brock ¹  Andreas H. Guse ¹  René Werner²  Lena-Marie Woelk²  Christian Lohr ³  Christine E. Gee ⁴ 

[1] Department of Biochemistry and Molecular Cell Biology, University Medical Center Hamburg-Eppendorf, 20246 Hamburg, Germany;Department of Computational Neuroscience, University Medical Center Hamburg-Eppendorf, 20246 Hamburg, Germany;Division of Neurophysiology, Institute of Zoology, University of Hamburg, 20146 Hamburg, Germany;Institute of Synaptic Physiology, University Medical Center Hamburg-Eppendorf, 20246 Hamburg, Germany;

关键词: Ca²⁺ imaging;    fluorescence microscopy;    live-cell imaging;    low signal-to-noise ratio;    deconvolution;    image restoration;   

DOI  :  10.3390/ijms222111792

来源: DOAJ

【 摘 要 】

Live-cell Ca${}^{2+}$ fluorescence microscopy is a cornerstone of cellular signaling analysis and imaging. The demand for high spatial and temporal imaging resolution is, however, intrinsically linked to a low signal-to-noise ratio (SNR) of the acquired spatio-temporal image data, which impedes on the subsequent image analysis. Advanced deconvolution and image restoration algorithms can partly mitigate the corresponding problems but are usually defined only for static images. Frame-by-frame application to spatio-temporal image data neglects inter-frame contextual relationships and temporal consistency of the imaged biological processes. Here, we propose a variational approach to time-dependent image restoration built on entropy-based regularization specifically suited to process low- and lowest-SNR fluorescence microscopy data. The advantage of the presented approach is demonstrated by means of four datasets: synthetic data for in-depth evaluation of the algorithm behavior; two datasets acquired for analysis of initial Ca${}^{2+}$ microdomains in T-cells; finally, to illustrate the transferability of the methodical concept to different applications, one dataset depicting spontaneous Ca${}^{2+}$ signaling in jGCaMP7b-expressing astrocytes. To foster re-use and reproducibility, the source code is made publicly available.

【 授权许可】

Unknown