Comparing synthetic refocusing to deconvolution for the extraction of neuronal calcium transients from light fields.
Comparing synthetic refocusing to deconvolution for the extraction of neuronal calcium transients from light fields.
复制标题
比较合成重新聚焦与从光场中提取神经元瞬变的反卷积。
DOI：
10.1117/1.nph.9.4.041404
复制
发表时间：
2022-10
期刊：
影响因子：
5.3
通讯作者：
Foust AJ
中科院分区：
文献类型：
--
作者：
Howe CL;Quicke P;Song P;Verinaz-Jadan H;Dragotti PL;Foust AJ
光场显微镜（LFM）可以快速、高效、体积成像的神经元活动与钙指标。从不同算法重构的体中提取钙瞬态信号时，其时间信噪比（tSNR）和空间约束不同。我们评估了钙荧光成像的两种光场重建算法的能力和局限性。我们在急性小鼠脑切片中获得了大量标记或细胞内填充红色钙染料CaSiR-1的神经元光场图像序列。我们比较了在有和没有全变分正则化的情况下，通过合成重聚焦和Richardson-Lucy三维反卷积重建的体积中提取的钙信号的tSNR和空间约束。合成重聚焦和理查森-露西反卷积都能在三维空间中分辨来自单细胞和神经元树突的钙信号。与合成重聚焦相比，增加反卷积迭代次数改善了空间约束，但降低了tSNR。与在匹配平面上获得的交错宽视场图像序列相比，体积光场成像并没有降低钙信号的tSNR。LFM能够对单细胞体细胞（体积标记）和树突（细胞内负载）中的钙瞬态进行高体积率、体积成像。对tSNR、空间限制和计算成本的权衡表明，合成重聚焦或反卷积哪种方法能更好地实现未来LFM钙成像应用的科学要求。
Light-field microscopy (LFM) enables fast, light-efficient, volumetric imaging of neuronal activity with calcium indicators. Calcium transients differ in temporal signal-to-noise ratio (tSNR) and spatial confinement when extracted from volumes reconstructed by different algorithms. We evaluated the capabilities and limitations of two light-field reconstruction algorithms for calcium fluorescence imaging. We acquired light-field image series from neurons either bulk-labeled or filled intracellularly with the red-emitting calcium dye CaSiR-1 in acute mouse brain slices. We compared the tSNR and spatial confinement of calcium signals extracted from volumes reconstructed with synthetic refocusing and Richardson–Lucy three-dimensional deconvolution with and without total variation regularization. Both synthetic refocusing and Richardson–Lucy deconvolution resolved calcium signals from single cells and neuronal dendrites in three dimensions. Increasing deconvolution iteration number improved spatial confinement but reduced tSNR compared with synthetic refocusing. Volumetric light-field imaging did not decrease calcium signal tSNR compared with interleaved, widefield image series acquired in matched planes. LFM enables high-volume rate, volumetric imaging of calcium transients in single cell somata (bulk-labeled) and dendrites (intracellularly loaded). The trade-offs identified for tSNR, spatial confinement, and computational cost indicate which of synthetic refocusing or deconvolution can better realize the scientific requirements of future LFM calcium imaging applications.