Molecular Devices SpectraMax GEMINI EM, Руководство оператора

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Instrument Description
SpectraMax GEMINI EM Operator’s Manual 1-11
Luminescence
In luminescence mode, no excitation is necessary as the species being mea-
sured emit light naturally. For this reason, the lamp does not
fl ash, so no back-
ground interference occurs. A dark estimate is done over a dark reference,
and multiple readings are averaged together into one reading per well.
You can choose the wavelength where peak emission is expected to occur. In
addition, multiple wavelength choices allow species with multiple compo-
nents to be differentiated and measured easily. In luminescence read mode, no
emission cutoff
fi lter is used. The default setting for luminescence is the “zero
order” position where the grating monochromator acts as a mirror which
re
fl ects all light to the PMT detector.
The GEMINI EM is a microplate spectro fl uorometer with photomultiplier
tube detection. Some luminescence applications, such as gene reporter assays,
may require a luminometer with photon counting detection for greater sensi-
tivity.
Functional
Description
The SpectraMax GEMINI EM is designed to be operated using SoftMax Pro
software running on a computer connected to the instrument. Stand-alone
functions are limited to setting and enabling temperature control and opening
or closing the microplate drawer.
The information contained in this section provides an overview of the instru-
ment capabilities. For a complete description of the modes of operation, how
to choose instrument settings, etc., refer to the
SoftMax Pro User’s Manual .
Modes of Operation
The SpectraMax GEMINI EM can perform two types of readings: fl uorescence
and some luminescence applications. Within each read type, the SpectraMax
GEMINI EM can perform four modes of operation: endpoint, kinetic, spec-
trum, and well scanning. Instrument setup parameters for each read mode are
discussed in the
SoftMax Pro User’s Manual.
Endpoint Mode
Select from one to four excitation/emission pairs in endpoint mode to obtain
fl uorescent (RFU) or luminescent (RLU) readings for each well of a micro-
plate. Endpoint data is reported as RFU or RLU.
Kinetic Mode
Kinetic analysis can be performed for up to 99 hours. The kinetic read interval
depends upon the instrument setup parameters chosen in SoftMax Pro. At the
end of a reading, the default kinetic reduction is RFU/second or RLU/second
for each well. Other kinetic reductions (e.g., milliunits/minute) are also avail-
able.
Kinetic analysis has many advantages when determining the relative activity
of an enzyme in different types of microplate assays, including ELISAs and
the puri
fi cation and characterization of enzymes and enzyme conjugates.
Kinetic analysis is capable of providing improved dynamic range, precision,
and sensitivity relative to endpoint analysis.