Bouwer James C, Mackey Mason R, Lawrence Albert, Deerinck Tom J, Jones Ying Z, Terada Masako, Martone Maryann E, Peltier Steven, Ellisman Mark H
National Center for Microscopy and Imaging Research, Center for Research in Biological Systems, University of California at San Diego, La Jolla, CA 92093-0608, USA.
J Struct Biol. 2004 Dec;148(3):297-306. doi: 10.1016/j.jsb.2004.08.005.
We describe the technique and application of energy filtering, automated most-probable loss (MPL) tomography to intermediate voltage electron microscopy (IVEM). We show that for thick, selectively stained biological specimens, this method produces a dramatic increase in resolution of the projections and the computed volumes versus standard unfiltered transmission electron microscopy (TEM) methods. This improvement in resolution is attributed to the reduction of chromatic aberration, which results from the large percentage of inelastic electron-scattering events for thick specimens. These improvements are particularly evident at the large tilt angles required to improve tomographic resolution in the z-direction. This method effectively increases the usable thickness of selectively stained samples that can be imaged at a given accelerating voltage by dramatically improving resolution versus unfiltered TEM and increasing signal-to-noise versus zero-loss imaging, thereby expanding the utility of the IVEM to deliver information from within specimens up to 3 microm thick.
我们描述了能量过滤、自动最可能损失（MPL）断层扫描技术在中压电子显微镜（IVEM）中的应用。我们表明，对于经过选择性染色的厚生物标本，与标准的未过滤透射电子显微镜（TEM）方法相比，该方法可显著提高投影和计算体积的分辨率。分辨率的提高归因于色差的降低，这是由于厚标本中大量的非弹性电子散射事件所致。这些改进在提高z方向断层扫描分辨率所需的大倾斜角度下尤为明显。该方法通过显著提高与未过滤TEM相比的分辨率以及与零损失成像相比的信噪比，有效增加了在给定加速电压下可成像的选择性染色样品的可用厚度，从而扩展了IVEM从厚度达3微米的标本中获取信息的效用。
