Nanoscale visualization of extracellular DNA on cell surfaces

• NCJ Number: 308299
• Author(s): Anita Olsen; Christopher J. Ehrhardt; Vamsi K Yadavalli
• Date Published: September 2020
• Length: 9 pages
• Grant Number(s): 2017-DN-BX-0186
• Agencies: NIJ-Sponsored
• Publication Type: Research (Applied/Empirical)
• Award: Nanoscale Imaging and Chemical Analysis of Extracellular DNA in Trace Evidence Samples

Annotation

In this article, the authors demonstrate the integration of optical microscopy with AFM for the visualization of topographical distribution of extracellular DNA using fluorescence imaging, and they suggest that this investigative technique will improve understanding of the biological role of extracellular DNA as well as be valuable to forensic investigations that require analysis of trace samples containing very few cells.

Abstract

Nanoscale analysis of extracellular DNA (eDNA) that is present on the surface of cells in trace biological samples can provide insight into the understanding of DNA transfer through touch, and thereby, the role of eDNA is a biologically and forensically relevant phenomenon. While various bulk scale tools and DNA analysis can be used to quantitatively obtain this information, obtaining a three-dimensional (3D) visualization of the eDNA can provide a unique look into the spatial and temporal dynamics at the cellular level. In this study, we show how atomic force microscopy (AFM) can be integrated with optical microscopy to visualize the distribution of surface associate eDNA at a single cell level. Using a nucleic acid fluorophore such as Diamond™ Dye, the surface eDNA can be observed and quantified using fluorescence microscopy. This informational channel can then be overlaid with surface topography and cellular elasticity to provide structural visualization. Finally, chemical force spectroscopy can be used to obtain the distribution of surface-associated eDNA on the cell surface at the molecular level. Such integrated techniques can enhance understanding of the biological role of eDNA, and can also be potentially valuable for investigating challenging trace samples, containing very few cells for various analyses. (Published Abstract Provided)