Department of NanoBiophotonics, Max Planck Institute for Biophysical Chemistry, Göttingen, Germany.
J Biophotonics. 2010 Jul;3(7):417-24. doi: 10.1002/jbio.201000038.
We compare the performance of video-rate Stimulated Emission Depletion (STED) and confocal microscopy in imaging the interior of living neurons. A lateral resolution of 65 nm is observed in STED movies of 28 frames per second, which is 4-fold higher in spatial resolution than in their confocal counterparts. STED microscopy, but not confocal microscopy, allows discrimination of single features at high spatial densities. Specific patterns of movement within the confined space of the axon are revealed in STED microscopy, while confocal imaging is limited to reporting gross motion. Further progress is to be expected, as we demonstrate that the use of continuous wave (CW) beams for excitation and STED is viable for video-rate STED recording of living neurons. Tentatively providing a larger photon flux, CW beams should facilitate extending fast STED imaging towards imaging fainter living samples.
我们比较了视频速率受激发射损耗（STED）和共聚焦显微镜在活神经元内部成像中的性能。在每秒 28 帧的 STED 电影中观察到 65nm 的横向分辨率，比共聚焦显微镜的空间分辨率高 4 倍。STED 显微镜能够分辨高空间密度的单个特征，而共聚焦显微镜则不能。在 STED 显微镜中可以揭示轴突受限空间内的特定运动模式，而共聚焦成像则仅限于报告总体运动。随着我们展示使用连续波（CW）束进行激发和 STED 对于活神经元的视频速率 STED 记录是可行的，预计会有进一步的进展。CW 束暂时提供更大的光子通量，应该有助于将快速 STED 成像扩展到更弱的活样本成像。
