Eppendorf AF2200 Plate Reader - Manual (Page 65)

Predefined methods
Eppendorf ® PlateReader AF2200
English (EN)
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6.2.7 Cell viability (CellTiter-Blue Assay)
Introduction
The CellTiter-Blue Cell Viability Assay provides a homogeneous, fluorometric method for monitoring cell
viability. The assay is based on the ability of living cells to convert a redox dye (resazurin) into a fluorescent
end product (resorufin). Nonviable cells rapidly lose metabolic capacity and thus do not generate a
fluorescent signal.
Default measurement parameters
• Excitation wavelength: 535 (25) nm
• Emission wavelength: 595 (35) nm
• Number of flashes: 25
• Settle time: 0 ms
• Mode: Top
• Lag time: 0 μs
• Integration: 20 μs
• Gain: "optimal"
• Plate definition: Eppendorf Cell Culture Plate 96/F, black/clear" or comparable plates
User defined method parameters
• The measurement is performed at 535 / 595 nm with default measurement parameters.
• Define the number of blank replicates (possible number: 1 – 6).
• Define the number of control replicates (possible number: 1 – 6).
• Define number of samples and sample replicates (max. 58, depends on number of blanks and controls).
• Define appropriate sample identifier (default settings are ID_1, ID_2, ..).
• Check if entered parameters are correct.
• Required plate layout: Outer wells are left empty by default. Pipette in a vertical order, starting with
blanks at position B2, controls are pipetted in subsequent wells, samples must be placed in the
following wells.
• Press Start .
• For method results and data evaluation principle see chapter method results and evaluation procedure .
The method is restricted to 96-well microplates.
Because of faster evaporation the wells lying on the outer sides of the plate are not included in
the evaluation.
 Please fill them with buffer or distilled water.