Research Institute Facilities By Christine Andrews, Karen Gogala & Marja Simpson.

Research Institute Facilities

By Christine Andrews, Karen Gogala & Marja Simpson

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

Horticulture Centre Laboratory & Trial Site

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

Dehydrating Oven & Cool Room

Operating range is +10oC to + 200oC

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

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

Steam Generator

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

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.au/research/weather/index.htm

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

Equipment – Research Lab

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

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

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

Scanning Electron Microscopy

• Creates magnified images by using electrons instead of light waves

• Shows 3D images at much higher magnification

• Samples prepared – sputter coater

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)

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.

Automontage Microscopy

• Perfectly focused 3D images

• Increased depth of field software

• Allows images of small insects almost as good as the specimen itself

GIS System

• Manages spatial data and associated attributes.

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

HPLC and GC/MS

• High Performance Liquid Chromatograph

• Gas Chromatograph coupled with a Mass Spectrometer

High Performance Liquid Chromatograph

• Chromatography—what is it?

• Liquid Chromatography

• Basic Operation

• Equipment used

• Types of Chromatography

• Applications for HPLC

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 phases:a 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.

Chromatography

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

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.

Liquid Chromatography

• Chromatogram

Basic Operation

Equipment used

• Shimadzu HPLC

Types of chromatography

• Adsorption

•Ion Exchange

Applications• Chemical separations• Identification• Quantification• Purification

• Cosmetics,energy,food,life sciences pharmaceutical, biomedical, drug detection and

identification.

Herbicides Carbohydrates in vegetables

Gas Chromatograph/Mass Spectroscopy (GC/MS)

• Gas Chromatograph

• Mass Spectrometer

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)

Gas Chromatography

• Schematic diagram

Mass Spectrometer

• Creates charged particles (ions) from molecules.

• Analyzes those ions to provide information about the MW of the compound and it’s chemical structure

• Many types of MS which allow wide range of analyses.

• GC/MS is the coupling of GC with MS

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)

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.

GC/MS

• Mass Spectrum Jamaican coffee

Shimadzu GC/MS