Virtual Event, Actual Science

Biophysical Society 66th Annual Meeting

Learn about innovative in vivo, in vitro, cellular, and computational techniques and technology for answering critical questions in biophysical research


Explore the latest advancements in fields ranging from the fundamental physical behavior of biomolecules to human health and disease.

These presentations cover:

  • Applications and foundations of SMLM and genomics imaging, the requirements to do successful experiments, and the technology behind the Bruker Vutara VXL; and
  • Investigating large and rough samples such as tissues and hydrogels using AFM, including how a newly developed hybrid of a motorized and piezo stage enables multi-region AFM probing over a large, rough sample area while providing additional correlative optical data sets.

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On-Demand Presentations

Title Speaker(s) Abstract Time (PST) Location
Single-Molecule Localization for Advanced Biological and Genomic Imaging: The Bruker Vutara VXL Winfried Wiegraebe, Ph.D., Product Manager Super-Resolution Microscopy, Bruker
Read More 10:30am -12:00pm Esplanade, Room 158
Application of Large Area Mapping AFM for Automated Structural and Mechanical Analysis of Developing Cells and Tissues
Giselle Fontes Evilsizer, Sales Applications Scientist, Bruker Nano Inc.
Read More 1:30pm - 3:00pm Esplanade, Room 157

Single-Molecule Localization for Advanced Biological and Genomic Imaging:
The Bruker Vutara VXL

Most sub-cellular structures are smaller than 300 nm. Because the size of these structures is below the classical resolution limit for fluorescence microscopy, we cannot use the specificity of this powerful technology to resolve them. The single-molecule localization technology (SMLM) implemented in the Bruker Vutara VXL overcomes this limitation. With an optical resolution of 20 nm, SMLM is uniquely qualified to address biological mysteries that require specific labeling as used in fluorescence microscopy but higher resolution than can be achieved with diffraction-limited microscopy. In addition to SMLM, the Vutara VXL implements imaging workflows for multiplexed chromatin tracing and smFISH applications. Thus the system allows imaging, resolving, and quantifying cellular structures, molecular machines, proteins, RNA, and chromosomal structures.
SMLM modulates the emission from dye molecules to prevent their point-spread-function (PSF) spatial overlap. Advanced fitting algorithms localize the position of each dye molecule with a resolution well below the optical resolution limit. The proprietary bi-plane technology of the Bruker Vutara VXL extends the traditional 2D fitting into the third dimension, far away from the coverslip. In STORM and PALM, a laser switches dye molecules actively on and off. Special imaging buffers create a similar blinking spontaneously in dSTORM. DNA-PAINT relies on oligonucleotides' transient binding, and genomic applications use sparse labeling and multiplexing.

The Bruker Vutara VXL brings these methods to every scientific lab. The instrument is small enough to fit on a classical lab bench. No darkroom is required. The easy-to-use workflow-oriented software includes everything needed to control the system, determine localizations, visualize the results, and apply advanced statistical analysis. Fluidics hardware controlled by the Bruker Vutara VXL enables data-rich multiplexing and genomics workflow.

This workshop discusses the applications and foundations of SMLM and genomics imaging, the requirements to do successful experiments, and the technology behind the Bruker Vutara VXL.

FOR MORE INFORMATION:

Find out more about the technology featured in this presentation or our other solutions for biological imaging:

 

Presented by Winfried Wiegraebe, Ph.D., Product Manager Super-Resolution Microscopy, Bruker

Application of Large Area Mapping AFM for Automated Structural and Mechanical Analysis of Developing Cells and Tissues

Active forces in biological systems define the interactions between single molecules, growing cells and developing tissues. Further development of novel biomaterials for tissue engineering is driven by the biomechanical and molecular cues provided to cells by their environment which are crucial parameters that influence motility, behavior, and the fate of progenitor cells.

AFM can be successfully applied for comprehensive nano-mechanical characterization of single molecules, cells and tissues, under near physiological conditions. Currently, the trend is to extend this by studying the mechanobiology of living cells while evaluating their structure and the interaction with their cell culture substrates. In particular, it is interesting to understand how cell behavior is driven by the cytoskeletal dynamics and cell mechanics in typical cell culture scaffold scenarios. We will introduce the concept of automated large area multiparametric characterization of densely packed cell layers and highly corrugated tissue samples, where full automation, smart mechanical sample analysis, multiple scanner technology, and optical integration is critical for data throughput and reliable correlative microscopy. We will discuss how these developments, in combination with advanced optical and super-resolution microscopy techniques, can overcome the inherent drawbacks of traditional AFM systems for characterizing challenging biological samples.

Cells adapt their shape and react to the surrounding environment by a dynamic reorganization of the F-actin cytoskeleton. We will demonstrate how cell spreading and migration in living KPG-7 fibroblasts and CHO cells, can be studied with high-speed AFM and associated with spatially resolved cytoskeletal reorganization events. We will further extend this with high-speed mechanical mapping of confluent cell layers, which in combination with optical tiling can be applied to automated analysis of large sample areas.

External mechanical stress is known to influence cell mechanics in correlation to the differences in actin cytoskeleton dynamics. As a tool for analyzing the complex cellular mechanobiology, we went beyond purely elastic models, and performed sine oscillations (up to 1 kHz, amplitude 5-60 nm) in Z while in contact with the surface to probe the frequency-dependent response of living fibroblasts. We will further discuss how to calculate the viscoelastic properties, characterized by the dynamic storage and loss modulus (E’, E’’) distribution in such samples.

In the past, investigating large and rough samples such as tissues and hydrogels using AFM was challenging due to the limited z-axis of the AFM. Using osteoarthritic cartilage as an example, we will demonstrate how a newly developed hybrid of a motorized and piezo stage enables multi-region AFM probing over a large, rough sample area while providing additional correlative optical data sets.

FOR MORE INFORMATION:

Find out more about the technology featured in this presentation or our other solutions for biological imaging:

 

Presented by Giselle Fontes Evilsizer, Sales Applications Scientist, Bruker Nano Inc.

Speakers

Winfried Wiegraebe, Ph.D.
Product Manager for Super-Resolution Microscopy, Bruker

Winfried Wiegraebe is the product manager for super-resolution microscopy at Bruker. He has close to 30 years of experience in advanced microscopy in biology - including AFM, FCS, confocal microscopy, and super-resolution microscopy. Before joining Bruker, Winfried managed the Stowers Institute for Medical Research microscopy infrastructure in Kansas City, MO, US, and built the imaging pipeline at the Allen Institute for Cell Science in Seattle, WA, US. He studied Physics at the Technical University in Munich, Germany, and did his Ph.D. studies at the Max-Planck Institute in Martinsried, Germany.

Giselle Fontes Evilsizer, Ph.D.
Sales Applications Scientist, Bruker Nano Inc.