PowerPointPrsentation

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LEAD THE WAY
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Company Details

• Established 1991
• Headquarters Glattbrugg, Zürich, Switzerland
• Branches MMI GmbH, Germany
• MMI Inc., USA
• Employees 25
• Represented in more than 65 Countries

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mmi Product range
mmi CellManipulator
mmi CellCut Plus
mmi ApplicationSupport
mmi Cellector
mmi Service Support
mmi SmartCut Plus
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Getting to grips with microscopic manipulation
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mmi CellManipulator

• Optical traps for ultra precise contact free
  manipulation of microscopic particles

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System Components

• Inverted Microscope CCD Camera High Precision
  xy-Stage IR Laser Intuitive Software Computer

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The Principle of Optical Tweezers

• hold particles in three dimensions trapping
  laterally (x-y plane) and
• axially (z-direction)

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How does the multibeam works
Cells in solution
Objective
Focusing lens
Laser 1064 nm 3.0 W
Galvo scanners

• Laser deflection with two fast galvanometric
  mirrors

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mmi CellManipulator System Components

• Inverted mircoscope
• Specific IR Laser, fully motorized xy-stage
• Software, Galvo Scanner, CCD Camera

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mmi CellManipulator Software functions

• Plug in Software, modular integration into mmi
  CellCut
• Live video presentation
• Scanning of samples
• Documentation of samples through screenshots
• Positioning of up to 10 beams
• Moving particles in the whole field of view
• Grouping beams for rotation or scaling
• Full control over all system components

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Moving Chloroplasts
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Main Applications

• Research in biological motors
• Study of cell-cell interactions
• Measurement of binding forces, elasticity or
  viscosities
• Cell Sorting, Cell Positioning
• Observation of cells by FCS
• Colloid and Interface Science
• Forming specific groups of cells
• In vitro fertilization IVF
• Cell Fusion in combination with mmi CellCut
• Microsurgery in combination with mmi CellCut

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Biological Motors

• Restoring force is proportional to the
  displacement out of the trap
• The bead is constantly moving with Brownian
  motion
• Whenever it leaves the center the restoring force
  pulls it back
• If a molecular motor pull the bead away from the
  center of the trap restoring force would be
  imparted to the bead and thus to the motor

Measurement of the 8-nm steps of Kinesin against
a 5-pN force
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Actin / Myosin Model

• Studie of interactions between filaments which
  are linked to microbeads
• Force measurement is possible

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How to use tweezers

• to investigate the interactions responsible for
  producing movements
• inside cells.
• Explaining the mechanism of contraction in
  skeletal muscle on a molecular level Aspects of
  Actin-Myosin interactions at a nanometer
  resolution can be measured.
• The proteins Actin and Myosin form the motor
  system which makes our muscles contract. As the
  porteins interact, myosin converts molecules of
  ATP into mechanical energy, thus producing
  movement.

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Experimental arrangement

• The ends of a filament of actin are each attached
  to a microbead
• each bead is hold with a separate beam
• thus the filament is pulled like a tight string
• When this string of Actin interacts with Myosin,
  the actin filament contracts
• By monitoring the position of the beads, the
  amount of force and movement a single myosin
  molecule produces as one ATP molecule is broken
  down, can be measured.

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Measurement of binding forces

• Measurements involving DNA Study elasticity of
  DNA strands
• Single molecule studies
• Measure the force required to separate
  antigenantibody bond
• Antigen-coated microsphere is trapped and pulled
  away from an antibody-coated surface
• Force applied by the tweezers to break the
  microsphere-coupled antigenantibody bonds is
  measured.

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Colloid and Interface Science

• measuring the elastic properties of polymers,
  including biopolymers
• Manipulation of membrane structures
• Measuring the polymers' responses to controlled
  forces
• Measuring the forces exerted on DNA by RNA
  transcriptase
• Position colloidal particles reproducibly
• Position and release otherwise isolated pairs of
  charge-stabilized colloidal spheres

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Observation of cells

• Fluorescence correlation spectroscopy (FCS)
  measurement
• Measuring effect of drugs in the cell
• Confocal Microscopy
• Taking pictures

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Cell Sorting

• Sorting and assaying of different cells, bacteria
  or organelles to different reservoires
• Study the behavior of single organisms
• Immunology Sorting/Isolation of T-Helper Cells
• Microbiology Sorting of fluorescence marked
  bacteria

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In vitro fertilization - IVF

• Sterile working because of no contact with
  needles
• Selection of individual sperm possible
• Selective working

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Cell Fusion

• positioning of cells for single-cell
  electroporation and fusion
• In combination with e.g. Calcium Imaging
  techniques

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Microsurgery

• manipulation for cellular microsurgery and laser
  induced fusion
• manipulation and dissection of plant cells

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mmi CellManipulator Advantages

• Simultaneous control of up to 10 particles
• Contact free and precise moving of selected
  particles inside the whole field of view
• Each beam can be moved individually
• Upgradable with mmi CellCut and mmi Cellector
• Optional combination of solid state or nitrogen
  laser
• Option Measuring of viscosities or measuring of
  binding forces to highest precision

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MMI CellManipulator - References
Others Eberhard Karls Universitaet Tuebingen
Germany, University of Teramo Italy, The
University of Calgary Canada, Department for
Civil Engineering Calgary Canada, Department of
Biochemistry and Molecular Biology University of
Nevada USA, Center for Integrative Genomics
University of Lausanne Switzerland
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4 Quadrant Measurement - 4QD
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System Components of the 4 quadrant detection
system

• MBPS Controller
• Optics AssemblyDetector Head

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The Principle of Operation

• Position detection
• quadrant detector photodiode is sectioned into
  four sections
• Each section acts as a separate photodiode
• Incident light is focused to a spot on the
  detector
• Each quadrant generates a current in proportion
  to the light on the section
• Relative magnitudes of the currents are used to
  determine the centre of the light spot
• Spot diameter in the range of 1 2 mm is
  optimal.

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Advantages

• Force Measurement included in MMI CellTools
  Software
• Two independent channels of positional feedback
  in two axes
• Positioning within a few nanometers is achieved
• Each axis of each channel has manual micrometer
  adjustment to allow a wide range of operation
• Dedicated PID controllers generate the feedback
  signals in each axis for each channel
• User selectable gain within 104 to 108 volts/amp
• Electronic offset capability in both axes for
  both channels

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