JoVE | Peer Reviewed Scientific Video Journal

TOPICAL COLLECTIONS

Single-Molecule Localization Microscopy for Nanoscale Visualization of Cellular Structures

Guest Editors

Geng Wang

Biomedical Engineering Department, Northwestern University

Dr. Geng Wang is a research associate at Northwestern University. He earned his PhD in Precision Instruments from Tsinghua University in 2018, followed by postdoctoral training at the University of Washington and the University of Illinois at Urbana-Champaign. His interdisciplinary research spans single-molecule localization microscopy (SMLM), nonlinear optics and imaging, and optical coherence tomography (OCT), with broad applications in biomedicine. Dr. Wang has published over 20 papers in peer-reviewed journals, including Light: Science & Applications, Advanced Science, and Laser & Photonics Reviews, along with one book chapter and 10 patents. Several of his innovations have been licensed and translated into industrial applications. He is a Co-PI on an NSF grant, a recipient of the NSF-AGEP Research Exchange Award, and has chaired scientific conferences, including SPIE Biomedical Applications of Light Scattering.

Vadim Backman

Biomedical Engineering Department, Northwestern University

Dr. Vadim Backman holds multiple esteemed positions at Northwestern University. He is the Sachs Family Professor of Biomedical Engineering and Medicine at the McCormick School of Engineering and Applied Sciences. Additionally, he serves as a professor of Medicine (Hematology/Oncology), Biochemistry, and Molecular Genetics at the Feinberg School of Medicine.  At the Robert H. Lurie Comprehensive Cancer Center, he is the associate director of Research Technology and Infrastructure and also leads the Cancer and Physical Sciences Program. Furthermore, Dr. Backman is the director of the Center for Physical Genomics and Engineering. He received his PhD in Medical Engineering from Harvard University and the Massachusetts Institute of Technology.  Dr. Backman conducts research at the intersection of physical and biological sciences to elucidate the function of the human genome and its dysregulation in disease. He develops new nanoscale imaging and computational technologies that facilitate elucidation, leading to novel methods for regulating global gene expression patterns and translating them into clinical applications for disease diagnostics and therapeutics.

Collection Overview

Single-Molecule Localization Microscopy (SMLM) encompasses a class of super-resolution imaging techniques, including PALM (PhotoActivated Localization Microscopy), STORM (Stochastic Optical Reconstruction Microscopy), and PAINT (Points Accumulation for Imaging in Nanoscale Topography), that enable visualization of subcellular structures at resolutions beyond the diffraction limit of light. By stochastically activating sparse subsets of fluorophores and computationally localizing their positions with nanometer precision, SMLM reconstructs high-resolution images of biological structures at scales down to ~10nm. This transformative approach was recognized with the 2014 Nobel Prize in Chemistry.

The Methods Collection aims to present a comprehensive and reproducible protocol for performing SMLM, including sample preparation, fluorophore labeling, microscope setup, image acquisition, and data reconstruction. SMLM has been instrumental in revealing the nanoscale organization of proteins, nucleic acids, and membrane structures, offering critical insights into processes such as synaptic transmission, signal transduction, and chromatin organization. By providing a visual, step-by-step guide, this collection will make SMLM more accessible to biologists with limited microscopy experience and facilitate its adoption in diverse research fields.