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DELFIA Instrument Options and Settings

Overview

DELFIA assays require a plate reader that is capable of TRF (time-resolved fluorescence) detection. This is a time-resolved fluorescence intensity assay. We recommend using a Revvity VICTOR™ Nivo, VICTOR™ X, EnSight™, EnVision™, or EnSpire™ Multilabel Plate Reader (view our instrument product pages). If you are using a plate reader manufactured by another company, you can use our Europium standard solution (catalog number B119-100) and perform a serial dilution in DELFIA Enhancement solution to determine the sensitivity of your instrument. We also offer Samarium standard solution (catalog number B115-100) and Terbium standard solution (catalog number C558-100) if you are interested in multiplexing.

  • Fluorometers (filter fluorometer) vs. spectrofluorometers/monochromators 
    We recommend a time-resolved filter-based fluorometer for DELFIA Europium assays. Very sensitive time-resolved spectrofluorometers may be suitable for certain assays. Most plate readers are capable of TRF-type assays - a flash lamp is needed for TRF analysis, which is an option on most plate readers. You could also use laser excitation on an appropriate EnVision instrument.
    • A TRF filter fluorometer plate reader utilizes bandwidth filters in front of the fiber optic for excitation and in front of the detector for emission wavelength choice. The grating-type spectrofluorometers are generally less sensitive than filter fluorometers, and loss in sensitivity can cause issues in highly sensitive assays such as TRF.
    • A TRF-capable spectrofluorometer utilizes a monochromator (usually an excitation and emission grating) to discriminate wavelength positions of study for fluorescence excitation and emission. Spectrofluorometers can be up to 1000-fold less sensitive than filter fluorometers, though spectrofluorometers may work in some DELFIA applications.
    • The new double-monochromator instruments claim to have almost similar sensitivity as the ‘original’ filter readers. These may be suitable for most DELFIA assays.
    • For cell-based assays, top illumination is recommended, as some clear-bottom plates may absorb excitation at 340 nm. Top illumination is naturally required with white and black plates.
  • DELFIA Instrument Settings when using DELFIA Europium 
    We recommend generic default instrument settings for all DELFIA assays. We always recommend you check with the manufacturer of your particular instrument to see if they have additional recommendations for DELFIA assays.

Table: Settings for Revvity instruments

  General  VICTOR Nivo  EnSight  EnVision  EnSpire  VICTOR X 
Excitation  320 or 340 wide-banded filter  320/75nm (HH35000947)  UV (TRF340) dug11 + wg320 filter, barcode 101 (HH34005010)  UV (TRF340) dug11 + wg320 filter, barcode 101 (2100-5010)  UV (TRF340) dug11 + wg320 filter, (2300-5010)  340/60 nm (built-in in VICTOR X4 and X5) 
Emission  615nm narrow filter bandwidth  615/8nm (HH35000935)  615nm (monochromator setting)  Europium 615/8.5 nm, barcode 203 (2100-5090)  615nm (monochromator setting)  616/8.5 nm (11440003) 
Dichroic    D400 (HH35000971)  n/a  LANCE/DELFIA D400 single mirror, barcode 412 (2100-4170)  n/a  n/a 
Delay  400µsec  400µsec  200µsec  400µsec  200µsec  400µsec 
Window  400µsec  400µsec  400µsec  400µsec  400µsec  400µsec 
Other settings    Measurement time: 500ms  Number of Sequential Windows: 1  Number of sequential windows: 1  Number of Sequential Windows: 1  Counting cycle: 1000µsec 
Flash energy: 100mJ  Number of Flashes: 100  Number of flashes: 100  Number of Flashes: 100  Flash energy mode: High 
    Time between flashes: 2000µsec     

 

  • DELFIA Instrument Settings when using DELFIA Terbium
    • Settings for a filter fluorometer:
      • Excitation: 340 nm
      • Emission: 545 nm
      • Delay: 500 µsec
      • Window: 1400 µsec
      • Cycle: 2000 µsec
      • Flash energy: high

        If you are working with Terbium and/or Samarium, please view the measurement information on our DELFIA multiplexing page.

 

For research use only. Not for use in diagnostic procedures.  

The information provided above is solely for informational and research purposes only. Revvity assumes no liability or responsibility for any injuries, losses, or damages resulting from the use or misuse of the provided information, and Revvity assumes no liability for any outcomes resulting from the use or misuse of any recommendations. The information is provided on an "as is" basis without warranties of any kind. Users are responsible for determining the suitability of any recommendations for the user’s particular research. Any recommendations provided by Revvity should not be considered a substitute for a user’s own professional judgment.