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Nanonics Hydra BioAFM: Correlating Optical Super-resolution with AFM

HydraBio

Nanonics, the leading provider of cutting-edge AFM technology, is proud to present its newest breakthrough in bio atomic force microscopy: the Hydra BioAFMTM featuring VISTATM (Vivid Imaging Soft Touch AFM) mode. This innovation offers ultrasensitive single pN force mapping combined with the benefits of super-resolution optical/fluorescent imaging. Live cell imaging with <100nm optical resolution and direct correlation with AFM topography is now possible. The finest structures, such as microvilli, can now be analyzed with on-line mechanics. The Hydra BioAFM, with its single or multi-probe capability, allows versatile optical integration with dual microscopes & side illumination. The system enables super-resolution methods like STED, PALM, STORM, and live cell NSOM with any available dye.

Imaging Live Cells with Optical Integration

Image MDCK

Topography of live MDCK cells imaged with VISTA showing microvilli

The HydraTM  BioAFM is a revolutionary BioAFM that enables complete optical integration allowing the researcher to use the traditional biology toolkit with advanced SPM functionality in both air and liquid.  Applicable integrations are: Integration with any upright or inverted microscope, confocal , fluorescence NSOM and Raman spectroscopy. The HydraTM incorporates an extremely sensitive tuning fork feedback mechanism allowing for measuring single pN’s of force.  Finally with the largest Z range available and multiprobe capabilities the Hydra opens new directions in Bio AFM imaging and nano-manipulation.

The Hydra BioAFM features VISTATM (Vivid Imaging Soft Touch AFM) – enabling live cell imaging with the highest resolution. Combine highest resolution force mapping with controlled and known point of contact.

VISTA Offers

  • Lice cell imaging with probes exhibiting Q factors in the thousands even in liquid
  • Single pN force measurement – the highest force resolution of any commercial AFM
  • Force spectroscopy – no Jump to Contact, allowing for investigating surface forces to the last nm from the membrane surface
  • Ultralong extended probe tips with large aspect ratio and ideal angle for minimal cell perturbation
  • 90?m Z range – the largest of any commercial AFM for imaging biological samples

True Optical Integration

The Hydra BioAFM is designed for all forms of optical microscopy. The Hydra is the only BioAFM to integrate with either upright or inverted microscopes and dual (4pi) microscopes. Furthermore, side illumination for light sheet microscopy is available. The Hydra BioAFM has rendered obsolete solutions such as shuttling a sample from your AFM back and forth to an upright microscope. All of this is achieved with no compromise in force sensitivity and resolution.

The Largest Versatility in Optical Configuration

  • Upright
  • Inverted
  • Light sheet side illumination
  • Combined 4 pi illumination (dual upright and inverted microscope)
  • Brightfield & darkfield
  • Phase contrast
  • Epifluorescence
  • TIRF
  • Confocal fluorescence
  • Raman and all forms of spectral Imaging
  • Non linear two, three photon, SHG and STED imaging

See the Whole Picture

  • Highly scattering tissue
  • Bacterial cultures in Agar
  • Opaque samples, such as microfluidic networks

Optical Overlay

Biological research requires a comprehensive view of the sample; by combining multiple perspectives a deeper understanding emerges.

The Hydra BioAFM features a correlated view

  • AFM overlaid with real-time CCD optical imaging
  • Complete optical access combined with glass AFMprobes provides for an unobstructed view
  • Quick and easy probe-sample placement
  • Quick selection of the area of interest for AFMimaging within the CCD image
  • Generate beautiful 3D collages, correlating opticalview with topographical and force information

Over the last decade, a new powerful set of farfield optical tools has emerged for optical imaging of biological samples below the diffraction limit. These Super-resolution techniques have opened new horizons in the understanding of key biological phenomena. The Hydra BioAFM is compatible with all optical modes of Super-resolution. It permits the integration of a dual microscope for STED. PALM and STORM with total internal reflection prisms and an upright microscope are readily allowed.

The Hydra BioAFM can combine AFM characterization with the following Super-resolution techniques:

  • STED
  • PALM
  • STORM

NSOM: A New Frontier in Live Cell Imaging

Near-field Scanning Optical Microscopy (NSOM) marks a paradigm shift in Super-resolution imaging. Live cell imaging with this powerful technique has been compromised in the past by stiff AFM cantilevers. Today Nanonics, with VISTATM, achieves the potential of live cell imaging with NSOM.

Open New Frontiers of Super-resolution Optical Imaging (<100nm)

  • Permits absorption and fluorescence
  •  Avoids the need for special dyes
  •  Correlates all optical contrast methods with 3D topography
  •  Allows highly confined zeptoliter illumination for fluorescence correlation spectroscopy (FCS) with physiologically relevant dye concentrations
  •  Enables AFM correlated FRET and dynamic FRAP with time resolutions previously unachievable
  •  Opens the door for the study of near-membrane phenomena with highly confined Z illumination

 

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