FluoroMax-3 v. 3.1 (3 Mar 2006)
Optimizing Data
5-14
Note:
The excita-
tion range is
240–600 nm; the
emission range
is 290–850 nm.
Correcting data
Introduction
Collecting accurate information about the fluorescent or phosphorescent properties of a
sample depends upon several factors:
•
Equipment specifications
•
Sample characteristics
•
Timing considerations.
To ensure that the spectra collected indicate the actual properties of the sample and not
external conditions, data often must be corrected. To
correct
data means to subtract in-
formation from the data not directly related to the properties of the sample.
Gratings,
detectors, and other spectrometer components have response characteristics that vary as
a function of wavelength. These characteristics may be
superimposed on spectra, thereby yielding a potentially
misleading trace. For accurate intensity comparisons,
such as those required for quantum-yield determinations,
response characteristics must be eliminated. Supplied
with the instrument are sets of excitation and emission
correction factors to eliminate response characteristics.
These files
1
,
xcorrect
and
mcorrect
, are included
with the software.
Items that may be convoluted into a
spectrum
Ways to remove these artifacts
Fluctuations caused by the light source
Monitoring lamp output using the
reference detector,
R
, and using the
signal ratio
S/R
to correct lamp profile
or temporal fluctuations
Influence of the sample holder
Running a blank scan (which is then
subtracted from the sample scan)
System hardware (e.g., optics, detectors).
Using radiometric correction factors
To use radiometric correction factors, either:
•
Select the ones supplied with the program, or
•
Select a set generated at your facility during or after acquisition, discussed in the
following section. Acquiring radiometric correction factors is explained in Chaper
8:
Producing Correction Factors.
Blank Subtraction
and
Dark Offset
functions are described in the on-line FluorEs-
sence™ help files.
1
Filenames include a three-letter extension. For the sake of clarity, we have omitted the extensions in this section. Refer to the
software manual for specifics regarding extensions.
Summary of Contents for FluoroMax-3
Page 5: ...FluoroMax 3 v 3 1 3 Mar 2006 v 15 Declaration of Conformity 15 1 16 Index 16 1...
Page 6: ...FluoroMax 3 v 3 1 3 Mar 2006 vi...
Page 18: ...FluoroMax 3 v 3 1 3 Mar 2006 Introduction 0 12...
Page 36: ...FluoroMax 3 v 3 22 Feb 2005 System Description 2 8...
Page 96: ...FluoroMax 3 v 3 1 3 Mar 2006 Maintenance 6 12...
Page 156: ...FluoroMax 3 v 3 1 6 Mar 2006 Automated Polarizers 10 22...
Page 208: ...FluoroMax 3 v 3 1 6 Mar 2006 Bibliography 14 4...
Page 210: ...FluoroMax 3 v 3 1 6 Mar 2006 Declaration of Conformity 15 2...