Synergy HT

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Synergy HT

• Multi-Detection Microplate Reader

Andrew Jones Labtech International
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SynergyHT

• Synergy HT Multi-Detection Microplate ReaderThe
  Synergy HT utilizes a unique dual-optics
  design combined with monochromator wavelength
  selection to provide uncompromised performance in
  absorbance, fluorescence and luminescence reading
  modes.

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SynergyHT
Combination of a µQuant and an FLx800
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SynergyHT
Synergy HT
Multi-Detection Microplate Reader

• For absorbance measurements, the design includes
• Xenon flash lamp for UV/Vis range
• Monochromator for fast and easy wavelength
  selection between 200 and 999 nm
• High quality fiber optics with a Silicon
  Photodiode as detector
• A reference diode signal to reduce background
  signal

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SynergyHT
Xenon Flash Lamp

• Continuous spectrum from UV to IR
• High optical output
• Low heat radiation
• Long life
• No warm-up required

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SynergyHT
Optical System - Absorbance
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SynergyHT
Optical System - Absorbance

• Monochromator based system
• No filters
• Continuous wavelength selection
• Spectral scans

gtgt Great flexibility regarding applications
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SynergyHT
Optical System - Fluorescence
Light Source
Excitation Cartridge
Emission Cartridge
PMT
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SynergyHT
Synergy HT
Multi-Detection Microplate Reader

• For fluorescence / luminescence determinations,
  Synergy HT uses
• Custom mapped fiber optic bundles for maximum
  signal transfer and collection
• Tungsten halogen light source for highest
  sensitivity
• PMT detection

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SynergyHT
Optical System - Fluorescence

• Cartridge Based Excitation and Emission Filters
• 4 filters per Wheel
• Enclosed in a Cartridge
• Cartridge easy to change
• Users can have different cartridges for different
  application groups.

gtgt Easy to use, easy switch to new applications
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SynergyHT
Optical System - Fluorescence
Excitation Fibres
Bifurcated Mapped Fibre Optics
Emission Fibres
Consistent, even illumination and capture of
Fluorescence
gtgt High quality of results
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SynergyHT
Dual optic system with two lamps
Xenon Flash Lamp
and
Tungsten Halogen Lamp
Tg
Relative Energy
Xe
100
250
1100
750
500
Wavelength
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SynergyHT
Synergy HT with TRF
Multi-Detection Microplate Reader

• For Time resolved fluorescence, the design
  includes
• Xenon flash lamp and the monochromator to select
  excitation wavelength
• When TRF is selected, an additional fiber optic
  bundle routes light from the monochromator to the
  excitation side of the fluorescence optics
• The standard Excitation filter wheel is replaced
  by a so called TR Block and the Tungsten Lamp
  is switched off
• An angled mirror within the TR Block diverts
  light directly into the top (or bottom)
  excitation probe for time resolved fluorescence
  measurements

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SynergyHT
Synergy HT
Speed
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SynergyHT
Synergy HT
Incubation, injection and shaking

• Incubation and shaking are standard features
• Ambient 4C to 50C
• 3 shaking speeds
• Optional 2 injectors
• gtgt Increased range of applications

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What applications?

• Absorbance
• Fluorescence
• Luminescence
• and
• 6 384-well microplate formats

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Absorbance - Automatic Enhanced Reading Mode
Ensures the highest quality readings
OD Measured 2.700 after 8 flashes
OD Measured 2.678 after 64 flashes
1 0.005
ERROR
NON ENHANCED
OD Measured 0.675
ENHANCED
OD Measured 0.923
OD Measured 1.234
WHY ?
Plate
8 Flashes
8 Flashes
8 Flashes
8 Flashes
2
3
4
1
64 Flashes
OPTICAL DENSITY
If less than 2.0 OD then on to next sample
If Greater than 2.0 OD then 64 more flashes, then
on to next sample
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Absorbance

• ELISA (Enzyme Linked Immuno-Sorbant Assay)
• ADME Toxicity Drug Screening
• Protein Quantitation
• Nucleic Acid Quantitation
• Protein Cell Aggregation
• Chemical Measurements
• Cell Viability, Proliferation Death
• Equipment calibration

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What applications?

• Absorbance
• Fluorescence
• Luminescence

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Fluorescence

• Fluoresence Intensity
• Standard fluorescence
• FRET (fluorescence resonance energy transfer)
• Macromolecular interactions
• Accessibility of molecules
• Flash Fluorescence
• Follow and quantitate fast events e.g. calcium
  uptake
• TRF (time resolved fluorescence)
• Background free

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Fluorescence Intensity

• Direct quantitation (NADH, NADPH)
• GFP Molecular Biology
• Fluorescent substrates (enzyme activity
  monitoring, Fluorescent ELISA )
• Immuno-fluorescence
• Labeled antibodies used to locate or quantitate
  antigens
• Labeling (DNA, protein quantitation)

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FRET (fluorescence resonance energy transfer)

• 2 fluorescent molecules
• One is the donor (blue)
• One is the acceptor (green)
• When both are very close to each other, FRET
  occurs. The emission of the donor is reduced and
  the emission of the acceptor is increased
• Allows quantitation of interactions at a
  molecular level

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FRET Applications?

• Receptor / ligand binding
• Detection of nucleic acid hybridization
• Membrane fusion assays
• Distribution and transport of lipids
• Protein folding

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Flash Fluorescence Calcium uptake
F
F
CA
F
F
F
F
F
F

• Fast kinetic events
• Requires to read just after the injection of the
  test molecule

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TRF (time resolved fluorescence)
y
t
i
gtgt No background light
s
n
e
t
n
I
Time (µs)
0
2
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TRF (time resolved fluorescence)

• Why Time Resolved Fluorescence?
• Low Background Fluorescence
• Works well in the presence of analytical
  interference
• Scattered light
• Short-lived background fluorescence
• Cell lysate assays (highly auto-fluorescent
  samples)
• BUT
• Relatively low signal
• Expensive reagent
• Instruments were previously too expensive

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What applications?

• Absorbance
• Fluorescence
• Luminescence

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Luminescence ATP assay applications?

• Hygiene
• If there is no ATP in a sample/surface, the
  sample/surface is sterile (food, pharmacy)
• Cell proliferation
• If ATP increases, cell number is probably
  increasing!
• Cytotoxicity Apoptosis
• If ATP is decreasing, likely cell number is too!
• Reporter gene assays

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Key Benefits and Features

• Microplate instrument control and data analysis
  software
• SIMPLICITY
• Gen5s user interface has a thoughtful,
  uncluttered design providing access to all major
  functions in a variety of ways to accommodate
  multiple user styles
• On-line Help, Wizard, Tutorials and Samples
  provide the most comprehensive set of learning
  tools ever

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Graphical toolbar
Movable tree menu
Traditional drop-down menus
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Key Benefits and Features

• EFFICIENCY
• Gen5s Welcome Screen provides quick launch
  buttons to key functions, links to recently used
  files and direct access to an extensive list of
  pre-programmed protocols and experiments
• The exclusive StepWiseTM Tools makes creating
  procedure and data reduction steps fast and easy

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Key Benefits and Features

• POWER
• StepWiseTM procedure steps available based on
  the connected reader (read, dispense, shake,
  delay, incubate, etc.)
• Data reduction definition steps allow for
  multiple analyses on one plate or several
  (multiple data sets, multiple transformations,
  multiple cutoffs validations, multiple kinetic
  results, etc.)

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Key Benefits and Features

• VERSATILITY
• With Data Views, the user can create exactly the
  data set to be displayed, reported or exported in
  each protocol. Or, use the Default Protocol data
  views each time a new experiment is created.
• Data can be output to Excel immediately, or to a
  PowerExport, File Export or printed report (or
  some combination of all output types!)

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Key Benefits and Features

• COMPLIANCE
• For compliance to 21 CFR Part 11, Gen5 Secure
  provides unsurpassed system, data and protocol
  protection
• Extensive and Flexible user group settings allows
  much more granularity in defining protected
  functions and permissions

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