Cell Volume (3D) Correlative Microscopy Facilitated by Intracellular Fluorescent Nanodiamonds as Multi-Modal Probes

• Main Authors: Neeraj Prabhakar, Ilya Belevich, Markus Peurla, Xavier Heiligenstein, Huan-Cheng Chang, Cecilia Sahlgren, Eija Jokitalo, Jessica M. Rosenholm
• Format: Article
• Language: English
• Published: MDPI AG 2021-12-01
• Series: Nanomaterials
• Subjects: correlative microscopy; 3D CLEM; volume imaging
• Online Access: https://www.mdpi.com/2079-4991/11/1/14

Three-dimensional correlative light and electron microscopy (3D CLEM) is attaining popularity as a potential technique to explore the functional aspects of a cell together with high-resolution ultrastructural details across the cell volume. To perform such a 3D CLEM experiment, there is an imperative requirement for multi-modal probes that are both fluorescent and electron-dense. These multi-modal probes will serve as landmarks in matching up the large full cell volume datasets acquired by different imaging modalities. Fluorescent nanodiamonds (FNDs) are a unique nanosized, fluorescent, and electron-dense material from the nanocarbon family. We hereby propose a novel and straightforward method for executing 3D CLEM using FNDs as multi-modal landmarks. We demonstrate that FND is biocompatible and is easily identified both in living cell fluorescence imaging and in serial block-face scanning electron microscopy (SB-EM). We illustrate the method by registering multi-modal datasets.