Opterra Multipoint Confocal

Resolving Nanoscale Structures on Living Cells

Until now, AFM imaging of microvilli structures had to be conducted on fixed cells to accommodate the higher imaging forces of traditional AFM tapping modes. Although a living cell’s underlying cytoskeleton could be seen, it was not possible to obtain the resolution needed to clearly observe the microvilli on the surface of softer, unfixed living cells. BioScope Resolve is able to capture high-resolution images of microvilli on the surface of delicate, living cells using the low imaging forces of PeakForce Tapping®.

high-resolution AFM imaging, microvilli, living cells, PeakForceQNM
Bioscope Resolve with PeakForce Tapping now enables high-resolution imaging of microvilli on living cells. (25 µm and 10 µm images of MDCK cells captured at 1 kHz PeakForce QNM. Images obtained by Dr. Hermann Schillers, University of Munster.)

 

"It was previously impossible to resolve the finest structures of a live cell like microvilli, but now with the improved PeakForce Tapping on BioScope Resolve I can image them easily in one hour."

 

Dr. Hermann Schillers
University of Munster, Germany

 

 

Real-Time Cell Topography, Modulus, and Fluorescence Observation

BioScope Resolve utilizes PeakForce QNM to enable collection of both topography and modulus data simultaneously, for studies such as imaging living cells that have fluorescently labeled actin filaments. Using synchronization to capture correlated AFM and fluorescence images, real-time changes in cell topography, modulus, and fluorescence can be observed


For example, if cells are exposed to the actin-disrupting drug Cytochalasin, a simultaneous decrease in modulus and disappearance of the fluorescence signal is observed. This fluorescence disappearance correlates to the breakdown of the cell’s cytoskeleton.  Additionally, a time-lapse movie of atomic force microscopy and fluorescence images can be created to graph the fluorescence and modulus changes over time.

correlated AFM and optical imaging, cell topography
Line-by-line synchronization of atomic force microscopy and confocal microscopy for true correlation of AFM and optical data.

Molecular High-Resolution Imaging

BioScope Resolve provides best-in-class, highest resolution molecular imaging on an inverted optical microscope. The system’s design seamlessly integrates the AFM head with light microscope instrumentation so that neither technology is comprised in any way. With its nanomechanical characterization, highest resolution topography mapping, and optical microscopy abilities, this next-generation BioAFM is even able to resolve the individual subunits of bacteriorhodopsin (bR) trimers.

bacteriorhodopsin, AFM image of lattice structure
PeakForce Tapping image of the bacteriorhodopsin (bR) membrane protein lattice structure taken on an inverted optical microscope. The inset shows a single particle averaging of the bR trimer. The green circle shows a single lattice defect. The blue circle

Most Quantitative Cell Mechanics and Molecular Force Spectroscopy Data

Only Bruker’s exclusive techniques, PeakForce QNM and FASTForce Volume combine to provide the widest range of ramp frequencies and the most quantitative property mapping for biological samples. Additionally, higher imaging speeds and automated measurement capability provide more data in less time, leading to faster time to publication.

 

Bruker’s new FASTForce Volume mode complements PeakForce QNM to provide the widest range of ramp frequencies.

  • FASTForce volume data acquisition, from 0.1 Hz to 300 kHz
  • pN level trigger forces for the most sensitive, highest resolution force distance curves for force spectroscopy
  • Widest ramp frequency range, when combined with PeakForce QNM, from 0.1 Hz to 1 kHz in liquid or 2 kHz in air
AFM ramp frequencies
Widest range of ramp frequencies for more data in less time
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