Transcript Research Laboratory Instrumentation

Research Institute Facilities
By Christine Andrews, Karen Gogala & Marja Simpson
Horticulture Centre Equipment
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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
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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
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Captures images with coloured video camera, Delta-T SCAN splashprotected flatbed scanner
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WinDIAS and Delta-T SCAN image analysis software analyse
images
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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
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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
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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
geographicallyreferenced
information.
HPLC and GC/MS
• High Performance Liquid
Chromatograph
• Gas Chromatograph coupled with a
Mass Spectrometer
High Performance Liquid
Chromatograph
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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
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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
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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