Widefield Two-Photon Excitation without Scanning: Live Cell Microscopy with High Time Resolution and Low Photo-Bleaching.
Widefield Two-Photon Excitation without Scanning: Live Cell Microscopy with High Time Resolution and Low Photo-Bleaching.
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DOI：
10.1371/journal.pone.0147115
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发表时间：
2016
期刊：
影响因子：
3.7
通讯作者：
McConnell G
中科院分区：
文献类型：
--
作者：
Amor R;McDonald A;Trägårdh J;Robb G;Wilson L;Abdul Rahman NZ;Dempster J;Amos WB;Bushell TJ;McConnell G
我们通过双光子激发演示了荧光成像，无需像 Hwang 及其同事之前描述的那样扫描生物样本，但具有增加的场尺寸和高达 100 Hz 的帧速率。在装载荧光 Ca2+ 指示剂 Fluo-4 AM 的原代大鼠海马神经元培养物中记录突触驱动的 Ca2+ 事件期间，我们观察到与单光子激发相比，光漂白大大减少。这种方法不需要对显微镜进行昂贵的添加，有望在需要高时间分辨率的工作中发挥作用。
We demonstrate fluorescence imaging by two-photon excitation without scanning in biological specimens as previously described by Hwang and co-workers, but with an increased field size and with framing rates of up to 100 Hz. During recordings of synaptically-driven Ca2+ events in primary rat hippocampal neurone cultures loaded with the fluorescent Ca2+ indicator Fluo-4 AM, we have observed greatly reduced photo-bleaching in comparison with single-photon excitation. This method, which requires no costly additions to the microscope, promises to be useful for work where high time-resolution is required.