WORLD
PRECISION
INSTRUMENTS
Instrumenting scientific ideas
SI-BF-100
Biofluorometer: Reliable, simple and affordable
zz
The new SI-BF-100 is an LED-based fluorometer
for life science applications. It is predominantly
used in one of two ways:
zz
zz
Stand-alone for probe-based applications
Connected to a microscope for
fluorescence imaging
It is ideally suited for ratiometric calcium
detection (FURA-2) and ATPase detection (via
NADH fluorescence). With up to seven LED
modules (wavelengths), the SI-BF-100 covers
many fluorometric applications in neuroscience
and cell biology.
Unique fiber optic
coupling probe
allows for highly
efficient transfer of
light and ease of
placement. Custom
probes fit your
existing systems.
Recent advancements in optics and LED
technology simplify ratiometric calcium imaging,
making this equipment more affordable. A
breakthrough in WPI patented technology
allows the SI-BF-100 to use wavelengths below
380nm and produce more light intensity at
those lower wavelengths. This technology
significantly cuts the cost of photometric
calcium imaging without sacrificing resolution
or quality.
zz
zz
zz
zz
03/08/2016
03/08/2016
LED light sources require less power, give
off less heat and are more compact and
affordable
Sampling rates up to 1kHz (1,000 ratios/s
max.). At lower speeds, signal averaging is
used for noise reduction.
The warm up time of less than one minute
is a dramatic improvement over the
common 20–60 minutes required by xenon
or mercury light sources
Two auto ranging photomultiplier inputs
for monitoring multiple wavelengths from
a single emission output that can be
comprised of any wavelength of light for
which an LED module is available
Heidelberg
A World Precision Instruments Company
zz
zz
Using a separate reference
channel, ultra-stable, continuous
ratio calculations automatically
compensate for LED intensity
drift–less noise and more
accurate measurements
zz Application-specific probes
available for existing tissue baths
and cuvette systems
zz Ratio noise is <0.05 peak to peak;
drift is less than 0.1 unit/hour
zz Replace the emission filter easily
or change the LED modules to
transform the SI-BF-100 into a
general purpose fluorometer for
many other applications
Can accept external light source for use of
specific wavelengths or intensities
BNC analog outputs (±10V) relate to the
emission intensities for both the individual
channels and a ratio
How it Works
Until now, calcium imaging systems
compensated for errors and noise introduced
by the complexity of their design. The systems
required mechanical filters and used expensive
xenon or mercury light sources. The beauty of
the SI-BF-100 is its simplicity. The elegance of
its design reduces the noise introduced into the
system and errors inherent in traditional designs.
Monochromatic LED light sources using WPI
patented technology eliminate the need for
complex and expensive white light sources and
filter wheels. Because the LED modules can be
pulsed, sampling frequencies up to 1,000 cycles
per second are possible.
The LED light source emits specific excitation
frequencies which travel through the probe.
The excitation light can be comprised of any
wavelength of light for which an LED module is
available. The probe returns a single emission
or dual emissions to one or two photomultiplier
inputs, which are independently filtered for
specific wavelengths. You can monitor multiple
wavelengths from a single emission output.
The SI-BF-100 LED light source makes this
ratiometric fluorometer more compact, energy
www.wpiinc.com
WORLD
PRECISION
INSTRUMENTS
Instrumenting scientific ideas
efficient and affordable. And, the low-power light source produces much less heat. It warms up in less than a minute!
This incredible design is not limited to calcium imaging either.
By simply replacing the emission filters in front of the photomultipliers with the desired wavelength filters, your SI-BF-100
becomes a general purpose fluorometer for any application
you can imagine.
The biofluorometer in this
setup is measuring calcium
transients from a muscle tissue
setup on an SI-MKB muscle
tester platform.
SI-BF-100 SPECIFICATIONS
Ca2+Excitation Wavelength...................................................................................................... 340, 380nm (others available)
Ca2+Emission Wavelength..................................................................................................................510nm (others available)
ATPase (NADH) Excitation Wavelength..........................................................................................360nm (others available)
ATPase (NADH) Emission Wavelength............................................................................ 470nm, 570nm (others available)
Fiber Optic Light Input/Output........................................................................................................................SMA terminated
Bandwidth.......................................................................................................................................................1000 ratios/second
Ratio Noise....................................................................................................................................................... < 0.1 peak to peak
Analog Output Range . ............................................................................. 0–10V (continuous, equivalent to a ratio 0–10)
Analog Output Impedance................................................................................................................................................... 100W
Power....................................................................................................12VDC, 0.5A, (universal power supply, 110/240VAC)
Warm Up Time................................................................................................................................................................ <1 minute
Dimensions............................................................................................................3.5"H x 17"W x 13"D (88 x 431 x 330mm)
Method
Description
WPI-Supported
Application
Single excitation/
Single emission
Sample is excited at one frequency of light. As a
result, a single (but different) wavelength of light
is emitted and directed to a photomultiplier tube
(PMT).
Calcium measurement
with Fluo-4
Excitation: 490nm
Emission: 510nm
Single excitation/
Double emission
Sample is excited at one frequency of light. As
a result, two separate wavelengths of light are
emitted. The emission light is directed to two
different PMTs which measure the light intensities
independently. The ratio of the emission (as a result
of each frequency) is used. Technically, TAMRA is
not required, but data are more consistent with its
inclusion.**
ATPase measurement with
TAMRA
Excitation: 360nm
Emission: 470nm (NADH),
570nm (TAMRA)**
Double excitation/
Single emission
Sample is excited as two different frequencies
of light flash alternately. As a result, a single
(but different) wavelength of light is emitted and
directed to a PMT. The ratio of the emission (as
a result of each frequency) is used to determine
calcium concentration.
Calcium measurement
with Fura-2
Excitation: 340nm, 380nm
Emission: 510nm
Double excitation/
Double emission
Sample is excited as two different frequencies of
light flash alternately. As a result, two separate
wavelengths of light are emitted. The emission light
is directed to two different PMTs which measure
the light intensities independently.
No currently supported
examples
**NADH is auto-fluorescent and indirectly measures ATPase activity. TAMRA is a secondary fluorophore
and corrects for movement artifacts and fluctuations in light intensity.
Heidelberg
A World Precision Instruments Company
SI-BF-100
SI-BF-100 Will Replace Expensive Light
Sources in Fluorescence Applications
Applications for the Biofluorometer are not
limited to use with the SI-H line of equipment.
The Biofluorometer can now be used as
an affordable light source for virtually any
fluorescence study. It can use high powered
LED modules for applications requiring a more
intense light source.
You can connect the Biofluorometer directly to
the epifluoresence port on your microscope
for fluorescence studies using a fiber optic
coupling.
The Biofluorometer offers many advantages
over the standard xenon and mercury light
sources typically used for fluorescence
microscopy.
For excitation, the Biofluorometer uses high
output LED modules with specific wavelengths
that are targeted for particular applications.
It also includes two photomultipliers and
the necessary filters to examine emissions.
Additionally, it performs the ratio calculations
and has outputs to monitor the emission
wavelengths and their ratio.
More Affordable
Xenon or mercury white light sources are
expensive, and their bulbs must be replaced
after about 700 hours of use. In contrast, highpower LED modules cost a fraction and last up
to 10,000 hours.
On this dual excitation
probe, the LEDs in the
center of the probe are
illuminated, while the
exterior ring of LEDs is
dark.
As an added bonus, LEDs require much
less power to output the same intensity of
monochromatic light, making them cheaper to
operate.
The new
Biofluorometer
with high output
LEDs includes the
light source, timing,
photomultipliers,
filters and capture
hardware in a single
case for less than
$20,000.
High Stability/Cool Operation
LEDs produce very little heat compared with the
alternatives. This improves the stability of the
output. Light intensity tends to fluctuate as the
temperature changes. While this is not nearly
as much of a concern for LED lights which are
highly stable and not nearly as hot as traditional
light sources, it still remains a consideration.
Any fluctuation in light is seen as noise on the
measured signal.
To compensate for LED intensity drift, the
Biofluorometer uses a separate reference
channel to monitor the actual excitation light
emitted. This information stream is fed back
into the system so that the excitation intensity
can be continuously adjusted, providing a very
stable light output.
This redundancy ensures that the light output
is ultra-stable. At the end of the day, that means
that you see less noise and end up with more
accurate measurements.
Instantly Available
Typical fluorescence light sources require
anywhere from 20 to 60 minutes to warm
up. This is not the case with LEDs, which are
instantly ready. The Biofluorometer is ready
when you are.
Compact
The Biofluorometer, which includes the light
source, the PMTs and filters, is a self-contained,
compact, benchtop unit.
Incredibly Flexible
When your application changes, swapping
LEDs and filters is a simple affair. Fluorescence
dyes are available for many applications, and
LED modules are available for almost as many
wavelengths. Now, you are only limited by your
own imagination.
The SI-BF-100 Biofluorometer is the perfect
choice for a high output, affordable light source
for fluorescence studies.
SI-BF-100
Heidelberg
A World Precision Instruments Company
Biofluorometer: Reliable, simple and affordable
Available for Use
with the Cell Tester
The size of a single cell is so small that up
until now fluorescence experiments have
required researchers to use high-power
external lights, because the standard
Biofluorometer probe is much too large to
use with a single cell. These expensive light
sources connected to the microscope to
project specific wavelength(s) of light through
the sample.
With the advent of the updated SI-BF-100
Biofluorometer, fluorescence studies of
single cells just became much simpler and
more affordable.
Wavelength
Epifluoresence
Port
Possible Applications
405nm
DAPI, Cascade Blue, Pacific Orange
430nm
AlexaFluor 430, CFP, Pacific Blue,
Calcium Yellow, Lucifer Yellow
470-480nm
AlexaFluor 488, GFP, DyLight 488,
FluoProbes 488, Cy2, NBD
500nm
Fluo-4, Oregon Green, Calcium
Green, YFP, Voltage-Sensitive
Dyes (DI-4-ANEPPS, DI-8-ANEPPS),
AlexaFluor 500, Rhodamine Green,
Fluorescein, BODIPY-FL, TrueRed,
FluorX, Sytox Green
530nm
TRITC, AlexaFluor 532, BODIPYTMR, JOE, HEX
550-560nm
dsRED, AlexaFluor 546, AlexaFluor
555, QSY 7, QSY 9, TAMRA,
Rhodamine Red, Calcium Orange,
Cy3, DyLight 550, FluoProbes 547,
Xytox Orange
580-590nm
mCherry, Calcium Crimson,
AlexaFluor 568, AlexaFluor 594,
AlexaFluor 610X, BODIPY-TR,
Texas Red, Cy3.5, DyLight 594,
FluoProbes 594
635nm
AlexaFluor 633, Cy5, BODIPY
630, DyLight 633, DyLight 560,
FluoProbes 647
670nm
AlexaFluor 660, AlexaFluor 680,
BODIPY 650, Cy5.5, DyLight 680,
FluoProbes 682
Emission Fiber
Excitation Fiber
W O RLD PRECISION IN STRU M EN TS, INC.
USA: International Trade Center, 175 Sarasota Center Boulevard, Sarasota FL 34240-9258 USA
Tel: 941-371-1003 • Fax: 941-377-5428 • E-mail: [email protected] • Internet: www.wpiinc.com
UK: 1 Hunting Gate, Hitchin, Hertfordshire SG4 0TJ England • Tel: 44 (0)1462 424700 • E-mail: [email protected]
Germany: Zossener Str. 55, 10961 Berlin, Germany • Tel: 030-6188845 • Fax: 030-6188670 • E-mail: [email protected]
China & Hong Kong: Rm 29a, No8 Donfang Rd., Pudong District, Shanghai 200120 PRC • Tel: +86 688 85517 • E-mail: [email protected]
Brazil: Conselheiro Nabias, 756 sala2611, Santos-Sao Paulo 11045-002 Brazil • E-mail: [email protected]