Howard Hughes Medical Institute, Janelia Farm Research Campus, Ashburn, VA 20147, USA.
Proc Natl Acad Sci U S A. 2012 Jan 3;109(1):22-7. doi: 10.1073/pnas.1109202108. Epub 2011 Dec 21.
The signal and resolution during in vivo imaging of the mouse brain is limited by sample-induced optical aberrations. We find that, although the optical aberrations can vary across the sample and increase in magnitude with depth, they remain stable for hours. As a result, two-photon adaptive optics can recover diffraction-limited performance to depths of 450 μm and improve imaging quality over fields of view of hundreds of microns. Adaptive optical correction yielded fivefold signal enhancement for small neuronal structures and a threefold increase in axial resolution. The corrections allowed us to detect smaller neuronal structures at greater contrast and also improve the signal-to-noise ratio during functional Ca(2+) imaging in single neurons.
在活体小鼠大脑的成像中，信号和分辨率受到样本诱导的像差的限制。我们发现，尽管光学像差会在样本中发生变化，并且随着深度的增加而增大，但它们在数小时内保持稳定。因此，双光子自适应光学可以恢复到 450 μm 深度的衍射极限性能，并改善数百微米视场的成像质量。自适应光学校正使小神经元结构的信号增强了五倍，轴向分辨率提高了三倍。这些校正使我们能够以更高的对比度检测到更小的神经元结构，并在单个神经元的功能 Ca(2+)成像中提高了信噪比。
