在完整小鼠大脑深处用 GCaMP6 标记的神经元的体内三光子成像。
In vivo three-photon imaging of activity of GCaMP6-labeled neurons deep in intact mouse brain.
机构信息
School of Applied and Engineering Physics, Cornell University, Ithaca, New York, USA.
Nancy E. and Peter C. Meining School of Biomedical Engineering, Cornell University, Ithaca, New York, USA.
出版信息
Nat Methods. 2017 Apr;14(4):388-390. doi: 10.1038/nmeth.4183. Epub 2017 Feb 20.
Abstract
High-resolution optical imaging is critical to understanding brain function. We demonstrate that three-photon microscopy at 1,300-nm excitation enables functional imaging of GCaMP6s-labeled neurons beyond the depth limit of two-photon microscopy. We record spontaneous activity from up to 150 neurons in the hippocampal stratum pyramidale at ∼1-mm depth within an intact mouse brain. Our method creates opportunities for noninvasive recording of neuronal activity with high spatial and temporal resolution deep within scattering brain tissues.
摘要
高分辨率光学成像是理解大脑功能的关键。我们证明，在 1300nm 激发下的三光子显微镜能够实现超过双光子显微镜深度极限的 GCaMP6s 标记神经元的功能成像。我们在完整小鼠大脑内约 1mm 深的海马锥体层中记录了多达 150 个神经元的自发活动。我们的方法为在散射脑组织深处进行具有高时空分辨率的神经元活动的非侵入性记录创造了机会。
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本文引用的文献
1
Sensitive red protein calcium indicators for imaging neural activity.
Elife. 2016 Mar 24;5:e12727. doi: 10.7554/eLife.12727.
2
three-photon microscopy of subcortical structures within an intact mouse brain.
Nat Photonics. 2013 Mar 1;7(3):205-9. doi: 10.1038/nphoton.2012.336.
3
Ultrasensitive fluorescent proteins for imaging neuronal activity.
Nature. 2013 Jul 18;499(7458):295-300. doi: 10.1038/nature12354.
4
Long-term dynamics of CA1 hippocampal place codes.
Nat Neurosci. 2013 Mar;16(3):264-6. doi: 10.1038/nn.3329. Epub 2013 Feb 10.
5
In vivo two-photon microscopy to 1.6-mm depth in mouse cortex.
J Biomed Opt. 2011 Oct;16(10):106014. doi: 10.1117/1.3646209.
6
Two-photon calcium imaging of evoked activity from L5 somatosensory neurons in vivo.
Nat Neurosci. 2011 Jul 10;14(8):1089-93. doi: 10.1038/nn.2879.
7
Functional imaging of hippocampal place cells at cellular resolution during virtual navigation.
Nat Neurosci. 2010 Nov;13(11):1433-40. doi: 10.1038/nn.2648. Epub 2010 Oct 3.
8
Deep tissue multiphoton microscopy using longer wavelength excitation.
Opt Express. 2009 Aug 3;17(16):13354-64. doi: 10.1364/oe.17.013354.
9
Third harmonic generation microscopy.
Opt Express. 1998 Oct 26;3(9):315-24. doi: 10.1364/oe.3.000315.
10
Infrared multiphoton microscopy: subcellular-resolved deep tissue imaging.
Curr Opin Biotechnol. 2009 Feb;20(1):54-62. doi: 10.1016/j.copbio.2009.02.008. Epub 2009 Mar 25.
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