2. General Description
18
Instructions for Use for INFINITE M1000 PRO No. 30064852 Rev. No. 1.0
2011-09
2.2.3 Luminescence
STOP
Caution
Switch on the instrument at least 15 minutes before starting a luminescence
measurement to ensure stable conditions for the measurement.
Glow Type Chemi- or Bioluminescence
The
INFINITE M1000 PRO
provides measurement of glow type chemi- or
bioluminescence. Glow type means that the luminescence assay glows much
longer than a minute. Luminescence substrates are available which provide
stable enough light output over hours.
As an example, luminescence can be measured to determine the activity of an
enzyme labeled compound (-peroxidase, -phosphatase). Light emission results
from a luminescence substrate being decomposed by the enzyme. Under excess
of substrate, the luminescence signal can be assumed to be proportional to the
abundance of the enzyme-labeled compound. As with enzyme-based assays,
control of environmental conditions is critical (temperature, pH-value).
For practical aspects of luminescence assays, see:
Bioluminescence Methods and Protocols, ed. R.A. LaRossa, Methods in
Molecular Biology 102, Humana Press, 1998
Flash Type Luminescence (with Injectors)
In flash-type luminescence assays, the measurement is only performed during
the dispensing of the activating reagent or after a short delay time.
Flash type luminescence is one of the measurement modes that can be
performed with injectors.
Note
The plate detection sensor is only active if one of the injectors is in use
(strips “injection” or “dispense”).
Note
During luminescence measurements, it is important to close the lid
which covers the syringes and bottles of the reagent system to minimize
background signal.
Dual-color Luminescence
Selected assays emit light of two different wavelengths at the same time.
For these assays, wavelength discrimination during luminescence detection may
be required.
Tecan luminescence filters are optimized for the Chroma-Glo
TM
Luciferase assay
system, for BRET and for BRET
2 TM
. Filters are built into the luminescence filter
wheel according to the demands of the applied assay:
•
‘Lumi Magenta’: wavelength range of 370 to 450 nm and 610 to 700 nm
•
‘Lumi Green’: wavelength range of 510 to 540 nm
•
‘Lumi Blue 1’: wavelength range of 370 to 480 nm
•
‘Lumi Green 1’: wavelength range of 520 to 570 nm
•
‘Lumi Blue’: wavelength range of 400 to 515 nm
•
‘Lumi Orange’: wavelength range of 550 to 630 nm
