Nikolenko Volodymyr, Nemet Boaz, Yuste Rafael
Department of Biological Sciences, Columbia University, New York, NY 10027, USA.
Methods. 2003 May;30(1):3-15. doi: 10.1016/s1046-2023(03)00003-3.
Two-photon microscopy has revolutionized life sciences by enabling long-term imaging of living preparations in highly scattering tissue while minimizing photodamage. At the same time, commercial two-photon microscopes are expensive and this has prevented the widespread application of this technique to the biological community. As an alternative to commercial systems, we provide an update of our efforts designing custom-built two-photon instruments by modifying the Olympus FluoView laser scanning confocal microscope. With the newer version of our instrument we modulate the intensity of the laser beam in arbitrary spatiotemporal patterns using a Pockels cell and software control over the scanning. We can also perform simultaneous optical imaging and optical stimulation experiments and combine them with second harmonic generation measurements.
双光子显微镜通过实现对高度散射组织中活体标本的长期成像，同时将光损伤降至最低，彻底改变了生命科学。与此同时，商用双光子显微镜价格昂贵，这阻碍了该技术在生物界的广泛应用。作为商用系统的替代方案，我们提供了有关通过改装奥林巴斯FluoView激光扫描共聚焦显微镜来设计定制双光子仪器的工作进展。使用我们仪器的新版本，我们通过普克尔盒和扫描的软件控制，以任意时空模式调制激光束的强度。我们还可以进行同步光学成像和光学刺激实验，并将它们与二次谐波产生测量相结合。
