Biotek Synergy LX, User Manual

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40 | Chapter 3 : Getting Started
Fluorescence Protocols
Before you create a Fluorescence protocol, define at least one fluorescence filter cube
on the reader; see page 26.
Plate: Tap the field to select a plate type. Options include standard 96- and 384-well plates
and Take3 if the plate has been configured on the reader (see page 46).
Blanks: Tap the field to open a plate matrix that corresponds with the plate type. Tap up to
12 wells to be identified as Blank and then tap Save . For a 384-well plate, the matrix is
displayed in quadrants; tap the 1-2-3-4 box to change the view.
Filter Set: If more than one fluorescence filter cube is defined on the reader, tap the field
to cycle through the available filter cube names. The EX/EM wavelength values (e.g.,
485/528) are displayed to aid in selection.
Gain: By default, when a plate read is initiated, the reader performs an AutoScale routine
(see below) to determine the optimum PMT gain setting for the wells to be measured.
Alternatively, tap to change AutoScale to Use Value and then enter a specific gain number
in the range of 25–255.
If AutoScale is selected, the reader measures the full plate (or the columns selected at read
time) as fast as possible, using the defined protocol parameters and a starting gain value of
40. If any wells overrange, the process is repeated using a lower gain value of 25. When all
wells return valid measurements, the well with the highest RFU value is measured again to
determine the optimum gain value that generates a target signal value for the dynamic
range used (scaled to 50,000 for Standard range or 1,000,000 for Extended range). The
optimum gain value is then used to measure all wells on the plate. It is also stored with the
results and will be included in the output to printer and/or USB flash drive.
Synergy LX User Manual
1501000N Revision A BioTek Instruments, Inc.