108
Recommended Modular Configurations
FL3-11
The
basic
Fluorolog
®
configuration is formed from
single-grating monochromators
in excitation and emission
positions,
a
T-sample
compartment, and a red-
sensitive photomultiplier. Add
any accessory now, or expand
your capabilities later. The
FL3-11 provides outstanding
sensitivity and performance at
the lowest price.
Nanolog
®
Alternate between the best in
scanning resolution and stray-
light rejection to the instantaneous
acquisition and spatial resolution
of an imaging spectrometer with
a CCD (or InGaAs array in the
IR). Our NanoLog
®
is the prime
example of this configuration,
specially optimized for analyzing
carbon nanotubes, quantum
dots, and other nanomaterials.
FL3-22
The ultimate in stray-light
rejection, the double-grating
monochromators in excitation
and emission positions are
perfect for highly scattering
biological samples like lipids
and proteins, or solids like
powders, semiconductors,
or phosphors. You also get
a bonus in sensitivity. The
additive grating design allows you to open the slits twice
as wide as for the same resolution you would get in a
single-grating monochromator. Standard center third slits
let you push the stray-light envelope even further.
FL3-11-MF
2
Switch
from
steady-state
measurement to picosecond lifetimes
with the optional MF2 automated
system as easily as clicking on a
mouse, without any realignment. MF2
is the fastest, most sophisticated
system for molecular dynamics as
you probe the microworld of energy
transfer, dynamic depolarization,
or an endless list of other time-
dependent applications. Take eight
frequencies simultaneously as fast as
10 ms per point.
Multiple, automated ports on the spectrograph, IR detectors, grating turrets—ask any Spex
®
Fluorolog
®
applications engineer
today to help you assemble your most versatile spectrofluorometers.
NanoLog
®
spectrofluorometer,
your best choice for analyzing
nanomaterials.
e
f
Near-IR emission spectrum
of singlet O
2
generated from
[Ru(bpy)
3
]Cl
2
in D
2
O.
5
FL3-TCSPC
For
time-domain
lifetime
measurements coupled with steady-
state fluorescence spectroscopy,
this configuration cannot be beat.
We incorporate TCSPC, with true
single-photon sensitivity, into the
Fluorolog
®
with multiple sources
as options, including solid-state
NanoLED sources, and spark
lamps for intense, wideband pulsed
light, as well as the standard xenon
CW lamp
100
Versatility
Fluorolog
®
Accessories
Fiber-optic platform
F-3000
Use this accessory for
remote-sensing from 250–
850 nm for samples that
cannot be placed in the
sample chamber.
Solid-sample holder
1933
The solid-sample holder is
designed for solids including
thin films, powders, pellets,
microscope slides, and
fibers. The holder consists
of a base upon which a
bracket, spring clip, and
sample-block rest.
Microscope interface
For recording fluorescence experiments under a
microscope, this accessory consists of a fiber-optic adapter
plus excitation and emission fiber-optic bundles that carry
the light-source to the microscope optics, and fluorescence
emission from the sample back to the Fluorolog
®
.
Chopper with Lock-in Amplifier FL-1069L
For improved data-acquisition in extended-IR applications,
try our chopper and lock-in amplifier accessory.
MicroMax 384
Microwell-plate reader
The MicroMax 384 can
rapidly scan hundreds of
tiny samples within minutes
automatically. Useful for
pharmaceuticals
and
nanomaterials, you can
determine the fluorescence
characteristics fast using
microwell plates with up to
384 wells.
Automated four-position
thermostated cuvette-holder
FL-1011
This cuvette-holder keeps up to four
samples at a constant temperature from
–10°C to +80°C. The temperature is
maintained by a liquid mixture pumped
through from an external circulating
temperature bath (F-1000/F-1001, not included). The holder
also includes a magnetic stirrer to mix a turbid or viscous
sample while positioned in the optical path. A dual-position
thermostated sample-holder (FL-1012) is also available.
Liquid-nitrogen Dewar FL-1013
To measure phosphorescence or
delayed fluorescence, samples
are often frozen at liquid-nitrogen
temperature to preserve the
fragile triplet state. A Dewar flask
is used to freeze and maintain the
temperature of the sample. The
sample is placed in a quartz cell,
and slowly immersed in the liquid-
nitrogen-filled Dewar. The Dewar is
on a pedestal within the Fluorolog
®
’s
sample compartment.
Automated single-position thermostated cuvette-
holder FL-1027
This cuvette-holder keeps a sample at a constant temperature
from –10°C to +80°C. The temperature is maintained by a
liquid mixture pumped through from an external circulating
temperature bath (F-1000/F-1001, not included). The holder
also includes a magnetic stirrer to mix a turbid or viscous
sample while positioned in the optical path.
Automated polarizers
The FL-1044 automated
L-format
polarization
accessory permits complete
control and calibration
of
your
polarization
experiments
from
the
computer keyboard. You
can automatically rotate
the polarizers to determine
VV, VH, HH, and HV
components. An optional
T-format configuration (FL-
1045) is also available.
6
64
Thermoelectric heater/
cooler F-3004
For heating and cooling
samples without external
circulating baths. You can
rapidly heat and cool your
fluorescent material through
a wide range of temperatures
using the Peltier effect. A
magnetic stirrer is included.
Stopped-flow
Accessory
The stopped-flow accessory
adds the dimension of
kinetics research to your
instrument, perfect for
analyzing
fluorescence
reactions on the millisecond
time-scale.
More accessories for the Fluorolog
®
:
Model
Item
F-3026
Standard-lamp correction factor kit
1914F
Thermoelectrically cooled R928P photomultiplier tube
1920
4 mL quartz cuvette with cap
F-3011
250 µL cylindrical quartz microcell adapter
F-3012
250 µL cylindrical quartz microcell (requires F-3011 adapter)
1925
4 mL quartz cuvette with stopper
1938
Set of 5 cut-on optical filters, 1’’ x 2’’
1939
Set of 5 cut-on optical filters, 2’’ x 2’’
1955
20 µL HPLC flow cell
F-1000/1001 External circulating temperature bath
F-3029
Integrating sphere for quantum yields
F-3023
Cryostat
FL-1001
Front-face viewing option
FL-1014
Sample-compartment electronics
FL-1030
Thermoelectrically cooled near-IR photomultiplier tube
QC-SK
Reduced-volume 1 mL cell, 5 mm x 5 mm, with adapter and magnetic stirrer
TRIG-15/25
Trigger accessory
Contact your Fluorescence Representative for an up-to-date list of new accessories to enhance your experiments.
No other company offers you the choice of time-domain or frequency-domain upgrades. Who else can supply the applications
support and service to get the full potential from your instrument? HORIBA Scientific has full applications laboratories in the
USA, Europe, and Asia, plus affiliates and representatives the world over. You can rest assured that you have the support you
expect only from HORIBA Scientific.
7
47
Real-World
Performance
Detecting fluorescence in highly scattering samples
With highly scattering samples, fluorescence signals may be
overwhelmed by stray or scattered light from the sample,
making quantitative and qualitative analytical determinations
impossible. However, a double-grating monochromator on
the emission side drastically improves stray-light rejection.
The rhodamine-B data below left compare the performance
of a single-grating and a double-grating system on the same
highly scattering sample: a thin monolayer of rhodamine-B
on a microscope slide. The sample was scanned in the
front-face fluorescence detection mode with our best single-
grating system, and then with a model with double-grating
monochromators on both excitation and emission. In plot
A, stray light from the sample masks the rhodamine-B’s
fluorescence. Plot B—measured using the double-grating
system— shows a well-defined fluorescence peak at 540 nm.
Carbon-tetrachloride data illustrate the unmatched stray-
light rejection of the Fluorolog
®
by revealing all four Raman
bands, at 350.7, 351.8, 353.6, and 357.7 nm for CCl
4
.
The excitation wavelength was 348 nm, and the bandpass
settings on the excitation and emission monochromators
were 0.5 and 0.7 nm, respectively. Narrow slit-widths and
the ability to step the monochromator in small increments
are critical in resolving the 350.7 and 351.8 peaks.
Whether you’re working in biochemistry or nanomaterials, measuring calcium-migration,
intermolecular distances, or laser crystals, the sensitivity and flexibility of a Fluorolog
®
spectrofluorometers will help you gather more information on more samples in a smaller amount
of time. When the focus of your research changes, so can the Fluorolog
®
, adapting modularly to
the demands of your work with upgrades and innovations. Here are just a few examples.
Emission scan in front-face mode of a monolayer of rhodamine-B fluorescence with
a (A) single-grating monochromator and (B) double-grating monochromator. Note the
improved resolution of the peak near 540 nm when the double-grating monochromator
rejects scatter from the sample.
The four peaks of the Raman spectrum of CCl4 are easily resolved with double-
grating monochromators in a Fluorolog
®
.
8
55
Synchronous scanning for characterizing complex
mixtures
The observed fluorescence spectrum of a complex mixture
often contains overlapping spectral features. Synchronous
scanning offers a solution to this problem by simultaneously
scanning the excitation and emission monochromators with
a constant offset between them (in units of wavelength or
wavenumbers).
provided by a variety of solid-state detectors, covering
different spectral regions, is available, as are choppers and
lock-in amplifiers for enhanced sensitivity. Only a Spex
®
Fluorolog
®
IR system includes these components as
integrated features.
Fluorolog
®
IR systems also have interchangeable gratings
and optional grating-turrets to enhance efficiency in the IR
region, giving Fluorolog
®
spectrofluorometers IR capabilities
unmatched by any other instruments.
The scan of a mixture of polynuclear aromatic hydro-
carbons (PAHs) compares a synchronous spectrum and a
conventional emission spectrum for a mixture of five PAHs.
The green line is the emission spectrum acquired on a
Fluorolog
®
system with constant-wavelength excitation.
When the sample is scanned synchronously (red line), five
individual components are resolved into unique sharp peaks.
Infrared fluorescence (CW and lifetime)
Our Fluorolog
®
systems can be equipped to detect IR
fluorescence, opening up totally new applications for
fluorescence spectroscopy. For example, manufacturers of
pharmaceuticals can employ IR fluorescence to identify toxic
agents. In the world of nanomaterials, IR fluorometry can
determine the composition of mixtures of single-wall carbon
nanotubes. Also, probes in the red avoid interference from
native fluorescence in the blue. An IR spectrofluorometer
must be equipped with a red -sensitive photomultiplier tube
(PMT) or solid-state detector whose response is effective far
into the IR region. For fluorescence detection at wavelengths
longer than 850 nm, there are two possible paths: the
simplest is to mount either a PMT or InGaAs array that takes
you as far 2.2 µm, depending on your choice; the alternative
Synchronous scan (red) versus conventional emission spectrum (green) of a mixture
of polynuclear aromatic hydrocarbons.
Normalized excitation (above) and emission (below) spectra from Nd-doped
phosphate laser-glass in the red to near-IR.
9
46
Real-World Performance, continued
Fluorescence from the singlet state usually occurs within a few
nanoseconds after excitation. Because triplet transitions are
more inhibited, the average phosphorescence-decay times
are longer, ranging from microseconds to seconds, offering
a longer observation period for monitoring reactions, viewing
effects of the local molecular environment on a sample, or
following changes in the hydrodynamic characteristics of
macromolecular systems.
In phosphorescence experiments, the Fluorolog
®
with the
FL-1040 dual-lamp housing—which includes a pulsed light
source—can excite your sample with synchronized user-
specified delay and sampling windows, and can record time-
resolved spectral data.
A delay permits acquisition of a phosphorescence spectrum
without fluorescence interference. This selectivity is particularly
important for samples in which the analyte can be overwhelmed
by strong fluorescence from extraneous materials. In the
matrix scan of an aqueous mixture of terbium-ligand and
europium ligand below (ligand = benzophenone-antenna
chromophore), the Eu-ligand luminescence (especially at
650 and 700 nm) decays faster (0.6 ms) than the Tb-ligand
luminescence (1.1 ms).
Low-temperature scans for enhanced fluorescence
One way to protect a sample from molecular collisions that
quench luminescence is to isolate the sample in a rigid matrix.
Thus, cooling with liquid nitrogen enhances the phenomenon
of fluorescence, even for an otherwise dormant sample. The
graph below compares the fluorescence spectra of pyrene
acquired at room temperature (upper) and at liquid-nitrogen
temperature (lower). The FL-1013 Dewar accessory was used
to chill the sample. As dramatically demonstrated in the lower
plot, the low-temperature technique intensifies fluorescence
emission for the pyrene, and sharpens peaks to reveal greater
structural detail. The superior resolution of a Fluorolog
®
double-grating system optimizes measurements under these
conditions.
Time-resolved data-acquisition also allows you to
acquire phosphorescence-decay curves and compute
phosphorescence lifetimes.
Time-gated matrix scan of an aqueous mixture of Tb-ligand and Eu-ligand.
Emission spectra for pyrene acquired at (top) room temperature, and (bottom) at 77 K.
10
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