High Performance Atomic Force Microscopy from Asylum Research Designed Specifically for Biological Applications

Topics Covered

Background
Uncompromised AFM on an Inverted Optical Platform
     AFM Is More Than Just Pretty Images
     The Best Force Measurements - from the Commercial Pioneer of Force Spectroscopy
     No Hole in Our Head
     Leadership in Force Spectroscopy and Force Mechanics
Light Up Your AFM
     Seamless Optical and AFM Integration
     Versatility from Above: View, Illuminate, Navigate
Control Your Sample Environment
Additional Options
Choose the MFP-3D-BIO
Real-time Optical Navigation
Cantilever Auto-Tune in Liquid
Real3DTM Imaging - View Your Data in Three Dimensions in Real Time as You Scan

Background

The MFP-3D-BIO provides the highest sensitivity and most accurate images and measurements possible on an inverted optical platform. The NPSTM closed loop nanopositioning sensors on all three axes ensure distortion-free images on samples as small as proteins and as large as cells - in both air and liquid. The MFP-3D measures the cantilever deflection to better than 20pm (8pm typical) without artifacts, making the MFP-3D ideal for force measurements such as unfolding single molecules or probing cell mechanics.

Uncompromised AFM on an Inverted Optical Platform

AFM Is More Than Just Pretty Images

From single molecule contour lengths to mammalian cell volumes, your published results are only as good as the data you acquire. The MFP-3D provides a truly quantitative three-dimensional map of your sample - a necessity for accurate measurements you can trust. Asylum's superior sensor and flexure stage technology coupled with our intuitive control software also gives you the ability to zoom and offset with a single mouse click - even from a previously captured image. Independent lateral (X-Y) and vertical (Z) positioning stages eliminate the crosstalk errors common to systems using tip-scanners. And the large Z range (up to 40µm) enables imaging of tall samples and cells, force curves on highly adhesive soft materials like gels, and other challenging applications. All standard AFM scanning modes are supported, including contact mode with lateral force, AC mode with phase, and advanced modes such as Dual ACTM, nanomanipulation, nanoindentation and more.

2.5µm scan of supported lipid bilayers (5nm tall) adsorbed onto mica and imaged in phosphate buffered saline.

The Best Force Measurements - from the Commercial Pioneer of Force Spectroscopy

Asylum Research pioneered commercialization of picoNewton-scale force spectroscopy with its first product, the Molecular Force Probe (MFP). Asylum continues that leadership to this day with the MFP-3D.

No Hole in Our Head

Some competitors literally have a hole through their AFM head. While permitting the use of some commercially-available optical condensers, the hole compromises the integrity of the AFM optical lever design and the stability of the AFM stage. This adds noise to their scans, degrades imaging resolution, and hinders quantitative force and friction measurements.

In contrast, the MFP-3D measures picoNewton forces and resolves the smallest features of your samples. The flexure-mounted optical lever mechanism moves as a single robust unit, virtually eliminating non-linearities, off-axis motion and interference fringes. Our custom-designed condenser column provides unobstructed top-down sample viewing and enables standard optical microscopy modes, including phase contrast.

The unfolding of fibril amyloid beta-sheets in algal adhesive reveals a distinct sawtooth pattern. The force curve was fitted to the worm-like chain (WLC) model (dotted lines) and a persistence length of 0.22nm was calculated. Data from Mostaert et al. (2006), J. Biol. Phys. 32(5):393.

Leadership in Force Spectroscopy and Force Mechanics

  • Simple setup, calibration, acquisition, display and analysis (any channel vs. any channel) with easy customization for advanced users.
  • Superior 8pm deflection noise (typical), 20pm guaranteed.
  • Accurate force measurements with no additional hardware required.
  • Users can choose between open loop sensored force curves for the ultimate in low noise performance or closed loop force curves for the most accurate velocity control.
  • MacroBuilderTM graphical macro language allows custom force measurements to be easily programmed, even by beginners.
  • Large vertical Z range (15µm standard, 40µm available) accommodates demanding applications such as adhesion measurements on cells and very soft gels.
  • Force Mapping enables mechanical measurements at a grid of points with automated extraction of material properties such as elastic modulus.
  • Force Clamping for single-molecule and bond-rupture force spectroscopy measures unfolding or rupture kinetics under constant force, allowing direct comparison to theoretical models.
  • Perpendicular drive, flexured nanoindenter (optional) provides accurate force and displacement control with superior resolution, sensitivity and precision.

Force map on fixed MRC-5 fibroblasts, 30µm. Deflection image (top image) shows location of selected force curves (middle graph). Automated force curve analysis generates an elasticity map (bottom image) which displays the measured modulus by color at each of the 1600 force curve points taken on the cells and substrate. The lamellipodia at the top center of the force map appear much softer than expected due to the presence of a second cell underneath.

Light Up Your AFM

Seamless Optical and AFM Integration

The MFP-3D-BIO sets the industry standard for combining optical and atomic force microscopies in a single integrated tool specifically designed for biologists. Our team of biologists and optical engineers have optimized the MFP-3D for use with the leading inverted optical microscopes to ensure that you get the maximum benefit and productivity from the combination of these powerful techniques. High numerical aperture, water immersion, and TIRF objectives are all accommodated. No other AFM can match the MFP-3D's performance for optical integration, high resolution imaging, quantitative force measurements, and environmental controls. All of the following optical microscopy techniques are supported:

  • Brightfield
  • Phase Contrast
  • Epifluorescence
  • Confocal Laser Scanning
  • TIRF
  • FRET
  • FCS
  • FRAP
  • Ion Indicators (e.g. Ca2+ response)

Multiply-labeled fibroblasts imaged in buffer using contact mode AFM (a), and fluorescence microscopy (b). The MFP-3D's standard overlay feature produced the composite image (c). Our proprietary IR filter blocks the AFM laser, enabling clean, full-spectrum fluorescence imaging - including far-red fluorophores.

MFP-3D's Exclusive X-Y Scanned-Sample Design is Essential and Enabling For:

  • Apertureless Near-field Microscopy (ANSOM)
  • Raman and Tip-enhanced Raman Spectroscopy
  • Tip-enhanced Fluorescence
  • Fluorescence Lifetime Imaging (FLIM)

Locate a cell with Zernike phase contrast (gray) or fluorescence, examine features such as cytoskeletal structures (red) or the nucleus (blue), then zoom in for high-resolution topography or force measurements with AFM (copper). Overlay optical data on AFM topography for 3D analysis and presentation (below).

Versatility from Above: View, Illuminate, Navigate

The MFP-3D extends the capability of the inverted optical microscope by allowing you to view transparent and opaque samples from above while scanning with the AFM. The integrated top-view optics enable in situ laser alignment and tip positioning - without removing the AFM head and without ancillary equipment. Our unique design also allows the top-view objective lens to double as a high-quality condenser for phase contrast illumination. Top- or bottom-view optical images can be used to navigate the tip to any feature on the sample, scan that area at the nanoscale with the AFM, and then select specific locations for force curves - easily and seamlessly. Optical images can be overlayed on AFM data, and captured for documentation and publication.

The optical path through the head of the MFP-3D-BIO utilizes a high-quality objective which can be used to view the top of opaque samples and to align the laser onto the tip. The objective can also be used as a condenser for brightfield or phase contrast illumination on transparent samples by using a series of lenses and mirrors to illuminate the sample. Using these optical elements, high quality illumination is achieved for observation with the inverted optical microscope objective while not undermining the fidelity of the optical lever or Z-flexure design of the AFM.

Control Your Sample Environment

  • Petri Dish Holder - Specially designed sample plate for imaging cultured cells and other biological samples in Petri dishes.
  • Petri Dish Heater - Petri Dish Holder with the additional capability for heating temperature-sensitive samples from ambient to 45°C.
  • Closed Fluid Cell - Sealed chamber with 10 inlet and/or outlet ports for exchange of liquid or gas media.
  • BioHeaterTM - Closed Fluid Cell for imaging in liquid between ambient and 80°C.
  • Fluid Cell Lite - Economical portless fluid cell. Ideal for individual users at multi-user facilities.
  • CoolerHeater - Heats and cools samples with a Peltier element. Continuous temperature control from -35°C to +120°C.
  • Humidity Sensing Cell - Measures humidity within a sealed sample cell.
  • iDriveTM - Simplifies imaging in liquid with cantilever auto-tuning and elimination of mechanical resonance peaks.

Additional Options

  • High-voltage PFM - Piezoresponse Force Microscopy for very high sensitivity, high bias and crosstalk-free measurements for piezoelectrics and biological systems.
  • ORCATM Conductive AFM - Provides current-voltage measurement capability.
  • MFP NanoIndenter - True instrumented nanoindenting for quantitative measurements with unprecedented accuracy.

Choose the MFP-3D-BIO

Asylum has made our full capability system easy to learn and use for novices while providing advanced functionality for the most experienced users. Don't compromise your research by choosing an "easy to use" system with limited capabilities. Choose the MFP-3D-BIO for both full capability and ease of use.

Start with the basics. Press one button in ModeMasterTM (top) and the AFM is configured for you, showing a simplified view with only the preset parameters you need for your experiment.

Take control of your AFM. Control the instrument precisely for more advanced experiments, specifying exactly what you want to do in our full-featured interface. Don't be held back by inflexible software.

Automate your work. For your most demanding experiments, access the full programmability of the MFP-3D AFM easily from our visual MacroBuilder panel. Spread your wings - no computer coding required!

Real-time Optical Navigation

Guide the tip to any location for AFM imaging, force mapping or individual force curves using any real-time optical input - brightfield, phase contrast, fluorescence, etc. This capability is of particular importance when working with functionalized tips or soft samples that preclude AFM scanning prior to force spectroscopy experiments.

40X phase contrast image of MRC-5 fibroblasts on a Petri dish. With a single click you can direct the tip (red dot) to any point, or select a new scan area (green box) within the scan range (red box).

Cantilever Auto-Tune in Liquid

Tuning the cantilever for AC mode takes only a single mouse click with Asylum's exclusive iDrive electromagnetic actuation.

Real3DTM Imaging - View Your Data in Three Dimensions in Real Time as You Scan

 

The MFP-3D's exclusive real-time 3D display allows you to view a realistic representation of your sample while the image is still being acquired. You can overlay the AFM phase or other data channels, rotate the image to any point of view, and adjust the scaling and lighting - all during the scan. This powerful capability is only available on Asylum Research AFMs.

Sequence of real-time 3D renderings of E. coli on glass showing structure and fimbriae, 5µm scan.

And once you've acquired your image our ARgyleTM image processing software helps you:

  • Render your data for journal-quality presentation and publication.
  • Control color height maps, lighting angle and intensity, dynamic data range scaling, interactive real-time rotation, zooming and panning
  • Create Quicktime® movies and 3D stereo anaglyphs
  • Overlay different AFM and optical channels. For example fluorescence or phase contrast can be mapped onto AFM topography to correlate the data/information

Source: Asylum Research

For more information on this source please visit Asylum Research

Date Added: Jul 26, 2006 | Updated: Jun 11, 2013
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