Laboratory of Chemical Pharmacology, Graduate School of Pharmaceutical Sciences, The University of Tokyo, Tokyo, Japan.
Neural Netw. 2010 Aug;23(6):669-72. doi: 10.1016/j.neunet.2010.02.004. Epub 2010 Feb 24.
Targeted patch-clamp recordings are a promising technique that can directly address the physiological properties of a specific neuron embedded in a neuronal network. Typically, neurons are visualized through fluorescent dyes or fluorescent proteins with fluorescence microscopy. After switching to transmitted light microscopy, neurons of interest are re-identified and visually approached in situ with patch-clamp pipettes. Here we introduce a simpler method for neuron targeting. With fluorophore-coated pipettes, fluorescently labeled neurons and the pipette tips are simultaneously imaged at the same fluorescence wavelength in the same microscope field, so that the neurons and even their neurites are targeted without suffering from chromatic aberration or mechanical complication in optics. We did not find that the coated fluorophores affected the electric properties of pipettes or neurons. The novel technique will be wildly available for pipette micromanipulation under online visual control.
靶向膜片钳记录是一种很有前途的技术,可以直接研究嵌入神经网络中的特定神经元的生理特性。通常,通过荧光染料或荧光蛋白在荧光显微镜下对神经元进行可视化。在切换到透射光显微镜后,重新识别感兴趣的神经元,并在原位用膜片钳管进行视觉接近。在这里,我们介绍一种更简单的神经元靶向方法。使用带有荧光染料的管,在同一显微镜视野中同时对荧光标记的神经元和管尖端进行成像,在同一荧光波长下,这样就可以对神经元进行靶向操作,甚至可以对其神经突进行靶向操作,而不会受到色差或光学机械复杂性的影响。我们没有发现涂层荧光染料会影响管或神经元的电学性质。这项新技术将广泛应用于在线视觉控制下的管微操作。