AU Biomaterials Design Lab:Equipment/LS55 PerkinElmer

Description

• Perkin Elmer LS 55 Fluorescence Spectrometer
• This is a 0.1nm resolution, compact spectrophotometer that uses the windows-compatible FL WinLab™ software that can measure fluorescence, phosphorescence and bio- and chemi-luminescence using a pulsed Xenon lamp source.

Purpose of use

Instrumentation The LS55 Perkin Elmer Fluorescence Spectrometer, 230V uses a pusled Xenon source to measure fluorescence, phosphorescence, chemi- and bio- luminescence. The FL WinLab™ software has different modes operation like Scan, TimeDrive, and Ratio Data Collection, increasing the versatility of the instrument. The excitation and emission monochromators can be used independently or simultaneously. The instrument also has the ability to prescan the sample to determine the excitation peak and maximum emission.

Included with the LS55 Perkin Elmer Fluorescence Spectrometer, 230V is a single-cell sample holder that can hold up to a 1cm pathlength cell and has the option of stirring. The automated polarizer has two filter wheels that have both a horizontal and vertical polarizing element. The polarizer is controlled by the software.

Applications

• Fluorescence, phosphorescence and bio- and chemi-luminescence
• 3D excitation/emission scans
• 3D synchronous and kinetic scans
• Flat samples, like TLC plates and electrophoresis gels, can be analyzed using the plate reader accessory
• Wavelength kinetics (both single and multiple wavelengths)
• Intracellular ion analysis

Location

• Beeghly 207
• American University
• Washington, DC
  • access to the laboratory is restricted; see Dr. Miller, Dr. Hartings or Dr. Fox for access.

Contact

• contact with questions or to schedule access via email
  • Dr. Matt Hartings
  • Dr. Abigail Miller

Use

• There is a on/off switch located on the left-hand side of the instrument's panel.
• When the switch is turned on, the neon lamp on the front lower left-hand side of the instrument is lit.
• It takes about 1 minutes to initialize.
• The instrument should be completely switched on before switching on the computer.
• Switch on the PC
• When Windows has completely loaded, start the FL WinLab software.
• After instruments calibrated and self checked, proceed to toolbar to select methods.
• Samples can be placed into the sample compartment.
• The door to sample compartment should be closed before samples are run.

Single Position Cellholder (Standard)

• This cell holder requires no alignment or set up.
• It holds 12.5mm square (10 mm pathlength) cuvettes and can be thermostatted using circulated water.

Stirred Single Position Cellholder

• This cell holder requires no alignment or set up.
• It holds 12.5mm square (10 mm pathlength) cuvettes and can be thermostatted using circulated water.
• The stirrer status can be set to low, high, or off by clicking the accessory icon.

Stirred Four-Position Chellchanger

• This cell holder requires no alignment or set up.
• It holds 12.5mm square (10 mm pathlength) cuvettes and can be thermostatted using circulated water.

Biokinetics Accessory

• This cell holder requires no alignment or set up.
• It holds 12.5mm square (10 mm pathlength) cuvettes and can be thermostatted using circulated water.
• The temperature sensor located in the block of the cellholder will report the temperature of the sample.
• After the sensor is calibrated using the FL WinLab LS55 Status application, the application will automatically record the temperature at the start of data collection in the result dataset header.
• Using the Read application, the intensity, polarization, or anisotropy data can be collected simultaneously with the temperature and saved as a spreadsheet.
• The event marker button on the front plate accessory can be used for identification of times when reagents are added to the cuvette.

Safety

• The equipment must be grounded.
• There are three Caution labels on the instrument, indicating that the user's manual needs to be consulted.
  • In the sample area, the caution label warns users of the potential for eye damage caused by staring into the beam in the sample compartment.
  • Near the mains inlet fuse, the caution label warns users that the operator replaceable fuse is a CSA approved fuse, not an IEC fuse.
  • At the front left of the instrument, by the hand screw that fixes the top cover, the caution label warns that removing the cover can expose hazardous voltages.
    • The caution label is coupled with a high voltage label.
• The other High voltage label is located inside the instrument on top of the monochromator cover.

Specifications

Spectral

• Wavelength range: 190 to 1100nm
  
• Spectral bandwidth: 1nm (190 to 1100nm)
  
• Wavelength accuracy: ±0.1nm
  
• Wavelength repeatability: ±0.5nm
  

Source

• Source: Xenon discharge lamp, equivalent to 20kW for 8μs duration
  
• Pulse Width at Half Height: <10μs
  

Detectors

• Sample Detector:
  • Gated photomultiplier with modified S5 response, max operation ~650nm
  • Red-sensitive R928 photomultiplier (optional), max operation ~900nm
• Reference detector: Photodiode, max operation ~900nm

Monochromators

• Monk-Gillieson type monochromators
  • Excitation:
    • Range: 200-800nm with zero order selectable
    • Spectral bandpass: 2.5-15.0nm, selected in 0.1nm increments
  • Emission:
    • Range: 200nm-650nm with standard photomultiplier with zero order selectable; 200-900m with optional R928 photomultiplier
    • Spectral bandpass:2.5-20.0nm, selected in 0.1nm increments
    • Filters:
      • Computer selectable cut-off (high pass) at 290, 350, 390, 430, and 515nm
      • A blank that acts as a shutter
      • A 1% T attenuator
      • Clear beam position
        

Phosphorescence Mode

• Variable delay and gate times
• Minimum total period: 12.0ms (50Hz)
  

Scanning speed

• Increments: 1nm
• Rate: 10-1500 nm/min
  

Sensitivity

• Signal to Noise: 500:1 rms
  • using the Raman band of water with excitation at 350 nm
  • excitation and emission bandpass: 10nm
    

Standard Cellholder

• A single position water thermostattable holder for 10mm cuvettes.

Physical

• Dimensions (W×D×H):
  

References

• LS 55 Fluorescence Spectrometer, 230V
• LS 55 Manual
• LS55 User Manual

Useful links

• AU Biomaterials Design Lab
• American University Chemistry

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