Research Institute Facilities

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Research Institute Facilities

• By Christine Andrews, Karen Gogala Marja Simpson

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Horticulture Centre Equipment

• Laboratory facilities with large working space
• Trial Site Area
• Plant Growth Cabinets
• Large Capacity Dehydrating Oven
• Cool Room
• Video and scanner based image analysis system
• Glasshouses with automatic heating and cooling
• Steam generator
• Automatic weather stations

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Horticulture Centre Laboratory Trial Site
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Plant Growth Cabinets

• 1 large cabinet, 2 smaller cabinets
• Temperature and humidity controlled
• Lighting intensity variable by switching lamps
• 24 h timers provide control between instruments

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Dehydrating Oven Cool Room

• Operating range is 10oC to 200oC

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Video and scanner based image analysis system

• Captures images with coloured video camera,
  Delta-T SCAN splash-protected flatbed scanner
• WinDIAS and Delta-T SCAN image analysis
  software analyse images
• Usage WinDIAS- Measurement of the area of
  healthy and diseased plant leaves
• Delta-T SCAN- Leaf
  measurement, Root length measurement, object size
  analysis, eg. soil particles, seeds, measurements
  from photographs or copies, count objects, eg.
  seeds

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Glasshouses

• Glasshouse size 3m x 7.5m x 3m
• Automatic cooling and heating system
• Winter heating capacity 10oC overnight, 20oC day
• Summer cooling 20-25oC
• Lighting is provided by incandescent and
  fluorescent lamps which is 24 h timer controlled

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Steam Generator

• Soil and plastic container treatment to
  control soil borne fungal diseases, nematodes and
  weeds

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Automatic Weather Station

• 2 stations
• Records Wind speed, wind direction, air
  temperature, rainfall, relative humidity, solar
  radiation, logger calculates evaporation
• Data available online http//www.orange.usyd.edu.a
  u/research/weather/index.htm

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Soil Shaker

• The Endecotts EFL 2000 is a vibrating shaker that
  is used to carry out sieve tests in conjunction
  with sieve stacks for particle sizing of various
  material samples.
• Sizes of sieves available 1.0mm
• 2.0mm
• 500micron
• 250micron
• 125micron
• 63micron

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Equipment Research Lab

• Atomic Absorption Spectrometer
• UV/Visible Spectrometer
• Scanning Electron Microscope
• Fluorescence Microscope
• PCR System
• Automontage Microscope
• GIS System
• GC/MS
• HPLC

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Atomic Absorption Spectrometer (AAS)

• Measures the amount of light absorbed by atoms
• Liquid sample aspirated, aerosolized mixed with
  gas
• Ignited in flame
• Atoms reduced to free state which absorbs light

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UV/VIS Spectrophotometer

• Measures amount of light a sample absorbs
• A beam of light passes through onto a detector
• Amount of molecules in a sample can be detected
• Both UV visible spectra

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Scanning Electron Microscopy

• Creates magnified images by using electrons
  instead of light waves
• Shows 3D images at much higher magnification
• Samples prepared sputter coater

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Fluorescence Microscopy

• Sample you want to study is the light source
• Energy absorbed by atom it gets excited
• Electron jumps to a higher energy level
• Drops back to ground state, emits a photon
  (fluorescing)

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PCR Room

• Polymerase Chain Reaction is a molecular
  biological technique for amplifying DNA without
  using a living organism.
• PCR is commonly used in medical and biological
  research labs for a variety of tasks.

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Automontage Microscopy

• Perfectly focused 3D images
• Increased depth of field software
• Allows images of small insects almost as good as
  the specimen itself

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GIS System

• Manages spatial data and associated attributes.
• It is a computer system capable of integrating,
  storing, editing, analysing, and displaying
  geographically-referenced information.

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HPLC and GC/MS

• High Performance Liquid Chromatograph
• Gas Chromatograph coupled with a Mass
  Spectrometer

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High Performance Liquid Chromatograph

• Chromatographywhat is it?
• Liquid Chromatography
• Basic Operation
• Equipment used
• Types of Chromatography
• Applications for HPLC

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What is chromatography?

• Chromatography colour and to write
• Originally described by Tswett in 1906 who
  devised a method to separate plant pigments using
  a tube filled with CaCO3.
• Basically it is a broad range of physical methods
  used to separate and /or to analyse complex
  mixtures
• Components to be separated are distributed
  between two phasesa stationary phase bed and a
  mobile phase which flows through the stationary
  bed.
• Individual species are retained by the stationary
  phase (packing) based on various interactions
  such as surface adsorption, relative solubility
  of the sample in the mobile phase and charge.

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Chromatography

• LC-mobile phase is a solvent and stationary phase
  is a packed bed of silica particles.

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Liquid Chromatography

• HPLC is this process conducted at a high
  velocity and under pressure.
• Sample can be in an aqueous form or in an
  organic/aqueous form.
• Sample is injected onto the column and is pushed
  through by the mobile phase(eluent) under high
  pressure.
• Components are retained and separated on the
  column.They elute at different times depending on
  their chemical interaction with the packing in
  the column.
• The time at which they elute (retention time) is
  a characteristic of that compound.
• After compounds elute,they enter a detector(PDA)
  which creates an electronic signal. The greater
  the concentration of the compound, the greater
  the signal.

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Liquid Chromatography

• Chromatogram

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Basic Operation
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Equipment used

• Shimadzu HPLC

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Types of chromatography

• Adsorption

• Ion Exchange

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Applications

• Chemical separations
• Identification
• Quantification
• Purification
• Cosmetics,energy,food,life sciences
  pharmaceutical, biomedical, drug detection and
  identification.

Carbohydrates in vegetables
Herbicides
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Gas Chromatograph/Mass Spectroscopy (GC/MS)

• Gas Chromatograph
• Mass Spectrometer

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Gas Chromatograph

• Mobile phase is an inert gas such as helium
• Sample is injected into a heated injection port,
  becomes vapourised and travels onto the column
  by means of the carrier gas.
• Column is made of fused silica onto which is
  coated the liquid stationary phase and it is
  enclosed in a heated zone(oven)
• Compounds become separated as they interact with
  the column
• Variables are temperature,gas flow, and column
  specifications.
• Separated compounds identified by specific
  detector.(FID.NPD,ECD)

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Gas Chromatography

• Schematic diagram

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Mass Spectrometer

• Creates charged particles (ions) from molecules.
• Analyzes those ions to provide information about
  the MW of the compound and its chemical
  structure
• Many types of MS which allow wide range of
  analyses.
• GC/MS is the coupling of GC with MS

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GC/MS

• A. Capillary column interface which connects
  GC to mass spectrometer
• B. Sample enters ionization chamber
• Ionization occurs. A beam of electrons impacts
  the sample molecules which lose an electron
  becoming positive (M)
• C. A positive potential is applied to repel the
  ions out of the ionization chamber and into the
  mass analyser.(filter)

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GC/MS

• Mass analyser separates the positively charged
  particles under vacuum according to their mass.
• Particles then enter a detector which sends
  information to the computer and resulting
  chromatograms give a mass spectrum of the sample
  components.
• Identification of the compound relies on the
  fact that every compound has a unique
  fragmentation pattern.

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GC/MS

• Mass Spectrum Jamaican coffee

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Shimadzu GC/MS