Biotek Synergy H1, Руководство оператора

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Filter Cube
Most Synergy H1 models are equipped with a filter cube that contains excitation and
emission filters, mirrors, and, if required, polarizing filters.
Excitation and emission filters are used for obtaining fluorescence and luminescence
measurements. The excitation filter selects the band of light to which the sample will be
exposed. The emission filter selects the band of light with the maximum fluorescence
signal of the sample, to be measured by the photomultiplier tube (PMT).
For filter-based, top-reading fluorescence analysis, the Synergy H1 uses mirrors to direct
the excitation and emission light paths. Mirrors are required for fluorescence polarization
(FP) measurements to direct light to the sample, because fibers cannot carry polarized
light. Mirrors also provide increased gain/sensitivity for fluorescence intensity (FI) and
time-resolved fluorescence (TRF) measurements. The filter cube stores up to two mirrors
and there are two possible mirror types:
l
A 50% mirror is a glass slide with silver dots. It works with any wavelength in the
range of 200 to 850 nm.
l
A dichroic mirror is wavelength-specific: It requires the excitation and emission
filters to fall within specific ranges. Dichroic mirrors provide better sensitivity than
50% mirrors, but they are dye-specific.
Filters and mirrors are stored in the filter cube as described in Filter Cube Overview
starting on page 44 . If you run different types of fluorescence or luminescence assays, you
can replace the entire filter cube with a different one; this is the BioTek-recommended
option. Alternatively, you can install different filters or mirrors in the cube. Contact BioTek
for information on purchasing additional cubes, filters, and mirrors.
The reader is delivered with a filter cube installed, and the reader’s
onboard software is configured with the filter and mirror values and
their locations in that filter cube. When Gen5 communicates with the
reader, it requests this information and stores the values in a Filter Cube
table.
It is critical that the values in Gen5 and onboard the reader
exactly match the contents of the installed filter cube.
If you
exchange the filter cube or modify its contents, you must update Gen5’s
Filter Cube table and send the new information to the reader; see
instructions on page
35 .
The filter cube is accessed through a hinged door in the front of the
instrument. Do not open the door to access the filter cube during
instrument operation! Doing so may result in invalid data.
Learn more about exchanging and modifying filter cubes in the Filters and
Mirrors chapter.
32 | Chapter 3: Getting Started
BioTek Instruments, Inc.