Prabhakar Neeraj, Peurla Markus, Shenderova Olga, Rosenholm Jessica M
Pharmaceutical Sciences Laboratory, Faculty of Science and Engineering, Åbo Akademi University, 20520 Turku, Finland.
Institute of Biomedicine, Faculty of Medicine, University of Turku, 20520 Turku, Finland.
Molecules. 2020 Dec 13;25(24):5897. doi: 10.3390/molecules25245897.
Correlative light and electron microscopy (CLEM) is revolutionizing how cell samples are studied. CLEM provides a combination of the molecular and ultrastructural information about a cell. For the execution of CLEM experiments, multimodal fiducial landmarks are applied to precisely overlay light and electron microscopy images. Currently applied fiducials such as quantum dots and organic dye-labeled nanoparticles can be irreversibly quenched by electron beam exposure during electron microscopy. Generally, the sample is therefore investigated with a light microscope first and later with an electron microscope. A versatile fiducial landmark should offer to switch back from electron microscopy to light microscopy while preserving its fluorescent properties. Here, we evaluated green fluorescent and electron dense nanodiamonds for the execution of CLEM experiments and precisely correlated light microscopy and electron microscopy images. We demonstrated that green color emitting fluorescent nanodiamonds withstand electron beam exposure, harsh chemical treatments, heavy metal straining, and, importantly, their fluorescent properties remained intact for light microscopy.
correlative light and electron microscopy (CLEM) is revolutionizing how cell samples are studied. CLEM provides a combination of the molecular and ultrastructural information about a cell. For the execution of CLEM experiments, multimodal fiducial landmarks are applied to precisely overlay light and electron microscopy images. Currently applied fiducials such as quantum dots and organic dye-labeled nanoparticles can be irreversibly quenched by electron beam exposure during electron microscopy. Generally, the sample is therefore investigated with a light microscope first and later with an electron microscope. A versatile fiducial landmark should offer to switch back from electron microscopy to light microscopy while preserving its fluorescent properties. Here, we evaluated green fluorescent and electron dense nanodiamonds for the execution of CLEM experiments and precisely correlated light microscopy and electron microscopy images. We demonstrated that green color emitting fluorescent nanodiamonds withstand electron beam exposure, harsh chemical treatments, heavy metal straining, and, importantly, their fluorescent properties remained intact for light microscopy.
相关光电子显微镜（CLEM）正在彻底改变细胞样本的研究方式。CLEM提供了关于细胞的分子和超微结构信息的组合。为了进行CLEM实验，应用多模态基准标记来精确叠加光学显微镜和电子显微镜图像。目前应用的基准标记，如量子点和有机染料标记的纳米颗粒，在电子显微镜下可能会因电子束照射而不可逆地淬灭。因此，一般来说，样本首先用光学显微镜进行研究，然后再用电子显微镜进行研究。一个通用的基准标记应该能够在保留其荧光特性的同时，从电子显微镜切换回光学显微镜。在这里，我们评估了绿色荧光和电子致密纳米金刚石用于CLEM实验，并精确关联了光学显微镜和电子显微镜图像。我们证明，发出绿色荧光的纳米金刚石能够承受电子束照射、苛刻的化学处理、重金属染色，重要的是，它们的荧光特性在光学显微镜下保持完整。
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