LS-AFM

Life Sciences AFM
For soft-sample applications

The LS-AFM is used in life sciences applications when an inverted optical microscope is required for locating cells or other bio-materials on a surface. The LS-AFM can be retrofitted to almost any inverted optical microscope, or it can be purchased with the AFMWorkshop inverted optical microscope.

Price Range*
$58,945.00 -
$145,500.00
* Prices vary depending on options purchased, importation taxes, and installation - training fees.

Click to Submit Inquiries or Questions

Description

 

LS-AFM Details

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Overview
Stage
EBOX
Software
Video Microscope
Probe Holder
Gallery
Modes
Options
Specifications
LS-AFM Overview
Available with AFMWorkshop inverted microscope Turnkey system with guaranteed results
Glass slides and petri dish sample holder No additional sample holding options required for most applications
Includes liquid scanner Readily scan samples in ambient air and liquids
Closed loop XY scanner Zoom to feature with accurate positioning for F/D curves
LabVIEW software with USB communication Readily adaptable to new operating systems
Probe exchange tool included Reduce time for probe exchange (& use any manufacturer's probes)
Includes top view video microscope Facilitates tip approach and laser alignment
Includes vibrating, non-vibrating, phase, LFM, and advanced F/D Most common scanning modes included for life sciences applications
Pricing
 
From $58,945.00
Download: LS-AFM Product Datasheet PDF
3-D model of LS-AFM
Measuring and Understanding Force-Distance Curves
 

The LS-AFM is used in biology applications in conjunction with an inverted optical microscope. Customers can buy the LS-AFM in two variations:

LS-AFM-A

For customers who already own an inverted optical microscope: In this configuration, AFMWorkshop fabricates a special plate that pairs the LS-AFM with the customer's existing inverted optical microscope.

LS-AFM-B

This configuration of the LS-AFM includes a fully-featured inverted optical microscope.

Features of the LS-AFM include:

  • Dry and Liquid Z Scanner
  • AFM Adapter Plate for Inverted Microscopes
  • Linearized XY Scanner
  • Advanced Force Distance Curves
  • Glass Slide and Petri Dish Sample Holder
  • Precision AFM Alignment System with Lock-Down
  • Included Modes: Vibrating, Non-Vibrating, Phase and LFM
  • Direct Drive Z Motor
  • Compatible With Standard AFM probes
  • Intuitive LabVIEW™ Software Interface
  • High Resolution Zoom Video Camera
  • High Resolution 24 Bit Scanning
  • USB EBox Interface
  • Available With AFMWorkshop's Inverted Optical Microscope (or Without)

 

 

LS-AFM Stage

The AFM Stage is secured on an adapter plate that is attached to the inverted optical microscope. There is an XY translation stage for moving the sample under the AFM Probe. Additionally there is an XY translation stage for moving the AFM over the inverted optical microscope axis.

 

LS-AFM highlighting features

 

 

 

Sample Stage for the LS-AFM

LS-AFM sample stage diagram

 

 

 

LS-AFM Diagram of Stage

 

 

Inverted Microscope (LS-AFM-B Only)

The LS-AFM may be purchased as an integrated AFM/Inverted Microscope. The Inverted Microscope includes all the options for Fluorescence, Phase Contrast, and standard Illumination imaging.

Included Items

  • Lamp Chamber for Florescence Microscopy
  • UV, V, B, G excitation Filters
  • Stage with 2" X 3" microscope slide translator
  • AFM Stage Adapter Plate(supplied by AFMWorkshop)
  • Objectives
    • Infinity LWD plan achromatic objective 10x/0.25 WD9.67
    • Infinity LWD plan achromatic objective 20x/0.40 WD7.97,
    • Infinity LWD plan achromatic objective 40x/0.60 WD3.76
    • Infinity LWD plan phase contrast objective 20x/0.40 WD7.97
  • Centering Telescope
  • DIC Polarizer
  • Lambda Plate
  • Bulb Cover
  • Phase Slide
  • C- mount port
  • Main Body

Not Shown

  • Power supply for florescence lamp
  • Power supply for illumination lamp
  • Video Camera

LS-AFM Stage

Back and side view of the LS-AFM stage without the AFM/ video microscope. The feet at the bottom may be removed if the stage is rigidly mounted to a surface.

 

 

LS-AFM EBOX

Electronics in the LS-AFM are constructed around industrystandard USB data acquisition electronics. The critical functions, such as XY scanning, are optimized with a 24-bit digital to analog converter. With the analog Z feedback loop, the highest fidelity scanning is possible. Vibrating mode scanning is possible with both phase and amplitude feedback using the high sensitivity phase detection electronics.

24 bit scan DAC

Scanning waveforms for generating precision motion in the X-Y axis with the piezo scanners are created with 24 bit DACS driven by a 32 bit micro controller. With 24 bit scanning, the highest resolution AFM images may be measured. Feedback control using the xy strain gauges assures accurate tracking of the probe over the surface.

Phase and Amplitude Detector Circuit

Phase and amplitude in the EBox are measured with highly stable phase and amplitude chips. The system can be configured to feed back on either phase or amplitude when scanning in vibrating mode.

Signal Accessible

At the rear of the EBox is a 50 pin ribbon cable that gives access to all of the primary electronic signals without having to open the EBox.

Precision Analog Feedback

Feedback from the light lever force sensor to the Z piezoceramic is made using a precision analog feedback circuit. The position of the probe may be fixed in the vertical direction with a sample-and-hold circuit.

Variable Gain High Voltage Piezo Drivers

An improved signal to noise ratio, as well as extremely small scan ranges are possible with the variable gain high voltage piezo drivers.


E-Box with features
Rear view of E-Box

LS-AFM Software

Software for acquiring images is designed with the industry standard LabVIEW™ programming visual interface instrument design environment. There are many standard functions, including setting scanning parameters, probe approach, frequency tuning, and displaying images in real time. LabVIEW™ facilitates rapid development for those users seeking to enhance the software with additional special features. LabVIEW also enables the LS-AFM to be readily combined with any other instrument using LabVIEW.

Prescan Window

Pre-Scan Window

A pre-scan window includes all of the functions that are required before a scan is started. The functions are presented in a logical sequence on the screen.

 

Scan Window

Scan Window

Once all of the steps in the pre-scan window are completed, the scan window is used for measuring images. Scan parameter, Z feedback parameters, and image view functions may be changed with dialogs on this screen.

 

Force/Distance Curves

Force/Distance Curves

There is a tab for measuring F/D curves in the AFMWorshop software. Data is exported to a .csv file for analysis in standard programs such as Microsoft Excel™.

 

LabVIEW programming window

LabVIEW Window

Industry standard programming environment. Readily customized and modified for specialized applications. Instrumentation already using LabVIEW can be added to the LS-AFM to create new capabilities.

 

 

Image Analysis Software

Included with the LS-AFM is the Gwyddion open source SPM image analysis software. This complete image analysis package has all the software functions necessary to process, analyze and display SPM images.


Image Analysis Software

 

  • visualization: false color representation with different types of mapping
  • shaded, logarithmic, gradient- and edge-detected, local contrast representation, Canny lines
  • OpenGL 3D data display: false color or material representation
  • easily editable color maps and OpenGL materials
  • basic operations: rotation, flipping, inversion, data arithmetic, crop, resampling
  • leveling: plane leveling, profiles leveling, three-point leveling, facet leveling, polynomial background removal, leveling along user-defined lines
  • value reading, distance and angle measurement
  • profiles: profile extraction, measuring distances in profile graph, profile export
  • filtering: mean, median, conservative denoise, Kuwahara, minimum, maximum, checker pattern removal
  • general convolution filter with user-defined kernel
  • statistical functions: Ra, RMS, projected and surface area, inclination, histograms, 1D and 2D correlation functions, PSDF, 1D and 2D angular distributions, Minkowski functionals, facet orientation analysis
  • statistical quantities calculated from area under arbitrary mask
  • row/column statistical quantities plots
  • ISO roughness parameter evaluation
  • grains: threshold marking and un-marking, watershed marking
  • grain statistics: overall and distributions of size, height, area, volume, boundary length, bounding dimensions
  • integral transforms: 2D FFT, 2D continuous wavelet transform (CWT), 2D discrete wavelet transform (DWT), wavelet anisotropy detection
  • fractal dimension analysis
  • data correction: spot remove, outlier marking, scar marking, several line correction methods (median, modus)
  • removal of data under arbitrary mask using Laplace or fractal interpolation
  • automatic xy plane rotation correction
  • arbitrary polynomial deformation on xy plane
  • 1D and 2D FFT filtering
  • fast scan axis drift correction
  • mask editing: adding, removing or intersecting with rectangles and ellipses, inversion, extraction, expansion, shrinking
  • simple graph function fitting, critical dimension determination
  • force-distance curve fitting
  • axes scale calibration
  • merging and immersion of images
  • tip modeling, blind estimation, dilation and erosion
LS-AFM Video Microscope

With the LS-AFM incorporates a top view optical microscope that is useful viewing of opaque samples and aligning the laser in the light lever. The inverted optical microscope is useful for locating cells and biomaterials for scanning.


LS-AFM 3 microscope views of ecoliThe LS-AFM presents three views of e. coli, from video optical microscope, inverted optical microscope, and atomic force microscope


Inverted Light Microscope

The AFM rests on a custom designe plate that facilitates direct viewing of the probe and sample with the inverted light microscope. With the inverted light microscope high resolution images of cells can be made, additionall a high resolution video image of the probe at the end of cantilever can be visualized.


Inverted Light Microscope

Inverted Light Microscope


Top View optical microscope

Included with the LS-AFM is a top view optical microscope with a 7:1 mechanical zoom tube, and a co-axial alumination source. The top view microscope is essenstial when using the LS AFM for imaging non-transparent samples.


Top View optical microscope

Top View optical microscope

LS-AFM Probe Holder

The LS-AFM utilizes a unique probe holder/exchange mechanism. Probes are held in place with a spring device that mates with a probe exchange tool. This combination makes changing probes fast and easy on the LS-AFM.


LS-AFM Probe Holder and Exchange

LS-AFM Image Gallery

The LS-AFM is designed for the most widely used types of measurements made with an AFM, including measuring F/D curves and imaging cells in a dry and liquid environment.

Measuring Stiffness of Biomaterials

Monitoring the deflection of a cantilever as it is pushed against a sample results in a force/distance curve. From the force distance curve many parameters may be measured, such as stiffness of the sample and probe-sample adhesion.

In biological samples, the most common application is measurement of intermolecular forces. For example, this could be used to measure the interaction force between an antigen and an antibody directly. Cell-cell adhesion forces and cellular stiffness can also be measured.

Advanced Force Distance Curve AFM software

The above screen shot demonstrates Advanced Force Distance Curve software measuring an AFM image.

1. Force-Distance data display region
2. Slider indicates the extension of the Z piezoelectric ceramic
3. Control parameter selection options
4. AFM Image for selecting locations for force-distance measurements

The Force/Distance Curve Measurement Software Interface includes all the features required for making advanced measurements. F/D curves may be made on single or multiple points of a sample surface. Control parameters include extend/contract rate, turn around trigger, and number of measurements per selected region.

Imaging Cells

Images of cells are readily scanned in both a liquid and dry environment with the LS-AFM. The inverted optical microscope facilitates direct placement of the probe on an area of interest for scanning. Additionally the inverted microscope can be operated in epifluorescence mode.

E. Coli cell and cheek cell by Atomic Force Microscope

Imaging cells in combination with an inverted optical microscope

The inverted optical microscope facilitates direct placement of the probe on an area of interest for scanning. Additionally the inverted microscope can be operated in epiflourescence mode.

Neutrophil A Cells

Life Sciences inverted optical microscope image of neutrophil -A cells

Inverted optical microscope image of neutrophil A cells. The dotted outline is the area scanned with the AFM.

Light shaded AFM image neutrophil-A cells

Light Shaded AFM image of the cells visualized in the optical microscope image.

 

Caco-2 Cells

Life Sciences AFM Caco-2 cells

Inverted optical microscope image of Caco-2 cells in the LS-AFM.

Clearly visible is the AFM cantilever on the right side of the image. A box identifies the area for AFM scanning.

3-D scale of Caco-2 cell 48 µm x 48 µm

3-D color scale image of the Caco-2 cell.

The scan range is 48 µm x 48 µm.

 

Epifluorescence and topographic AFM image of Caco-2 and quantum dots

CACO-2 cell structure in the presence of low concentration of quantum dots.

Left: Epifluorescence, showing brightfield (red), DAPI (blue), 2.2nm quantum dot PL emission at 560nm (green).

Right: Topographic AFM image of the indicated area.

LS-AFM Modes

Modes included with the LS-AFM:

Non-Vibrating (contact) In non-vibrating mode the deflection of the cantilever is held constant as the sample is scanned. This mode is used for hard samples, and for training purposes.
Vibrating (tapping) A piezoelectric ceramic is used to vibrate the cantilever at resonance. The amplitude of vibration is held constant as the sample is scanned. Both soft and hard samples are scanned with vibrating mode.
Phase While scanning in vibrating mode and holding the cantilever vibration amplitude constant, the phase shift between the drive signal and photo-detector are displayed. The phase image gives a map of the relative hardness at a sample's surface.
Frictional Force With the 4 segmant photodetector in the TT-2 AFM the L-R signal can be captured and displayed while scanning.
Advance F/D The advanced forc distance modes software allows force mapping, making curves from a non-feedback position, and controlling the inbound and outbound rates.
Dunk-n-Scan With the dunk-n-scan liquid cell, images of samples submerged in liquids can be made.

Z Scanner for Liquid Imaging


Diagram of liquid imagingDunk-n-Scan cell for liquid imaging of samples


Optional LS-AFM Modes:

The following modes may be purchase as options with the LS-AFM.

Conductive AFM (C-AFM)

Magnetic Force Microscopy (MFM)

Lithography

Scanning Thermal Microscopy (SThM)

Scanning Tunneling Microscope (STM)

Electric Force Microscopy (EFM)

LS-AFM Options

XY Sample stage

The optional base with XY translator gives added flexibility to the LS-AFM. By removing the inverted light microscope, noise floors as low as 100 picometers are achievable. The XY translator range is 12 X 12 mm with a resolution of 1 micron. At the top of the XY translator are magnets for holding standard AFM sample disks.


SA with translator



Vibration Solution

Inverted Light Microscope




Performance of the LS-AFM is greatly improved with the acostic enclosure combined with an active vibration table. The option includes:

- Acoustic Cabinet
- Active Vibration Table
- Base


Modes

The following modes are available for the LS-AFM:

Conductive AFM (C-AFM)

Magnetic Force Microscopy (MFM)

Lithography

Scanning Thermal Microscopy (SThM)

Scanning Tunneling Microscope (STM)

Electric Force Microscopy (EFM)

LS-AFM Specifications

40 Micron XY Scanner
Type Modified Tripod
xy Linearity < 1%
xy Range > 40 μm
xy Resolution < 3 nm closed loop
< 0.3 nm open loop
xy Actuator type Piezo
xy Sensor type Strain Gauge


16 Micron Z Scanner / Probe Holder
Includes both Air, and Dunk And Scan
Noise < 0.2 nm
Strain Gauge Resolution 1 nm
Tip Angle 10 °
Z Linearity < 5%
Z Linearity-Sensor < 1%


Light Lever AFM Force Sensor
Probe Types Industry-Standard
Probe Insertion Manual
Probe Exchange Tool
Probe Holding Mechanism Clip
Vibrating Mode Piezo
Electrical Connector to Probe
Laser/Detector Adjustment Range +/- 1.5 mm
Adjustment Resolution 1 µm
Minimum Probe to Objective 25 mm
Laser Type 670 nm Diode, < 3 mW
Laser Focus < 25 μm
Probe sample angle 10°

Detector
Type 4 Quadrant
Band Width > 500 kHz
Signals Transmitted TL, BL, TR, BR
Gain Low, High Settings

Digital Data Input Output
Connection USB

Scanning DAC
Number 2
Bits 24
Frequency 7 kHz

Control DAC
Number 2
Bits 14
Frequency 2 kHz

ADC
Number 8
Bits 14
Frequency 48 kHz

Z Motion
Type Direct Drive
Range 25 mm
Drive Type Stepper Motor
Min. Step Size 330 nm
Slew Rate 8 mm/minute
Limit Switch Top, Bottom
Control Software – Rate, Step Size


Analog Electronics
» Vibrating Mode
Freq Range 2 kHz – 800 kHz
Output Voltage 10 Vpp
Demod. Freq TBD

» Z Feedback
Type PID
Bandwidth > 3 kHz
Sample Hold Yes
Voltage 0-150 V

» XY Scan
Voltage 0 – 150 V
Bandwidth > 200 Hz
Pan & Zoom 22 Bits

» Tip Approach Cutoff < 20 μm sec.


Software
Environment LabVIEW
Operating System Windows
Image Acquisition Real Time Display
(2 of 8 channels)

Control Parameters
PID Yes
Setpoint Yes
Range Yes
Scan Rate Yes
Image Rotate 0 and 90°
Laser Align Yes
Vibrating Freq. Display Yes
Force Distance Yes
Tip Approach Yes
Oscilloscope Yes
Image Store Format Industry Standard
Image Pixels 16 x 16 to 1024 x 1024
H.V. Gain Control XY and Z
Real Time Display Line Level, Light Shaded,
Grey Color Palette
Calibration System Window
Probe Center  

Video Microscope

 
Minimum Zoom
Maximum Zoom
 
Field of view
2 x 2 mm
300 x 300 μm
 
Resolution
20 μm
2 μm
 
Working Distance
114 mm
114 mm
 
Magnification
45X
400X
 


Computer

  • Industry-standard Computer & Monitor (laptop available upon request)
  • Windows
  • AFMWorkshop LabVIEW.exe installed

* Z Noise performance depends greatly on the environment in which the LS-AFM operates. Best Z noise performance is obtained in a vibration-free environment. Contact AFMWorkshop for more information on our vibration isolation equipment and recommendations.

*Z Noise on the inverted microscope is <1nm.

**Every effort is made to present accurate specifications within this document. However, due to circumstances beyond the control of AFMWorkshop, specifications are subject to change without notice.

LS-AFM Product Datasheet PDF

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