WPI SI-BF-100 User manual

SI-BF-100
Biofluorometer for Fluorescence Imaging
World Precision Instruments
INSTRUCTION MANUAL
Serial No._____________________
061413
www.wpiinc.com

SI-BF-100
World Precision Instruments iii
Copyright © 2013 by World Precision Instruments, Inc. All rights reserved. No part of this publication
may be reproduced or translated into any language, in any form, without prior written permission of
World Precision Instruments, Inc.
CONTENTS
ABOUT THIS MANUAL ..................................................................................................................... 1
INTRODUCTION.................................................................................................................................. 1
Warnings.......................................................................................................................................... 3
Parts List ........................................................................................................................................... 3
Unpacking ....................................................................................................................................... 3
INSTRUMENT DESCRIPTION.......................................................................................................... 4
Front Panel ...................................................................................................................................... 4
Back Panel....................................................................................................................................... 6
Setting Up the System ................................................................................................................. 6
Changing the PMT Filters........................................................................................................... 7
Adding an LED Module............................................................................................................... 9
Changing the Sample Averages of the Display................................................................11
Changing the Operating Mode ..............................................................................................11
Configuring the System.............................................................................................................12
Modifying the Sampling Frequency................................................................................12
Modifying the Gain Adjust .................................................................................................12
Modifying the Photomultiplier Gain Voltage...............................................................13
Modifying the Filter Frequency.........................................................................................13
Setting up LED Modules......................................................................................................14
OPERATING INSTRUCTIONS.........................................................................................................15
Changing the Display ................................................................................................................15
Parameters Display...............................................................................................................15
Ratio/Sample Averages Display.......................................................................................16
Turning On/Off the Modules...................................................................................................16
Configuration Parameters for Various Fluorophores......................................................17
MAINTENANCE..................................................................................................................................17
Monitoring the LED Output.....................................................................................................17
ACCESSORIES.....................................................................................................................................18
SPECIFICATIONS................................................................................................................................18
APPENDIX A: EXAMPLES OF CALCIUM MEASUREMENT..................................................19
Intracellular Calcium Concentration Measurement ........................................................19
Calcium Measurements with Fluo-4.....................................................................................19
Calcium Measurements with Indo-1 ....................................................................................21
Calcium Measurements with Fura-2.....................................................................................22
APPENDIX B: EXAMPLE OF ATPASE ACTIVITY MEASUREMENT.....................................23
INDEX ...................................................................................................................................................26
WARRANTY ........................................................................................................................................27

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SI-BF-100
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ABOUT THIS MANUAL
The following symbols are used in this guide:
This symbol indicates a CAUTION. Cautions warn against actions that can
cause damage to equipment. Please read these carefully.
This symbol indicates a WARNING. Warnings alert you to actions that can
cause personal injury or pose a physical threat. Please read these carefully.
NOTES and TIPS contain helpful information.
Fig. 1—The Biofluorometer is rack mountable.
INTRODUCTION
The SI-BF-100 is an LED-based fluorometer for life science applications. It is ideally
suited for detecting changes in calcium concentration and ATPase activity in cells.
With the capability of being configured with up to seven LED modules at a variety
of wavelengths, the SI-BF-100 can be utilized in many fluorometric applications in
neuroscience, muscle physiology and cell biology.
NOTE: LED modules, filters and probe configurations described in this manual are
examples of what can be accomplished with this incredibly flexible Biofluorometer.
For custom configurations, please contact WPI at [e-mail].
Until now, biofluorometers were complicated devices with powerful light sources,
filter wheels that selected the proper wavelengths of excitation light, multiple
photomultipliers tuned to detect emissions from bound and unbound indicators and
processors that compensated for fluctuations in light intensity, as well as motion
artifacts. Now, by using LED modules, the SI-BF-100 is designed to have greater
control over the fluorescence measurement cycle. The frequency, duration and
firing sequence of each LED can be programmed through controls on the front
panel of the SI-BF-100. Additionally, you can set the intensity of the LED output, the
sensitivity and filter frequency of the photomultiplier (PMT), the gain of the amplifier,
and the sampling average to optimize the emission signals that can be recorded.
The unit also has a feedback mechanism to monitor the actual light output of the
Biofluorometer and make required adjustments to the output readings.

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The Biofluorometer can be used for many applications, limited only by your
imagination. Some example applications are provided in the appendices:
• Specific information on calcium measurement with several examples is located
in. “Appendix A: Examples of Calcium Measurement” on page 19.
• Information on using NADH/NAD and TAMRA to measure ATPase activity can
be found in “Appendix B: Example of ATPase Activity Measurement” on page
23.
The table belows outlines the example applications used in this manual.
Excitation Wavelengths/Emission Filters for Biofluorescent Dyes and Molecules
Dye or
Molecule
Excited
Active
Molecule
Assayed
Excitation
Wavelength
(nm)
Emission Filter Wavelength
and Range for PMT1
Emission Filter Wavelength
and Range for PMT2
Center, Range
(nm) Filter Number
Center, Range
(nm) Filter Number
Fura-2 Calcium 340, 380 510, 84 84-081
FLUO-4/Fura
Red
Calcium 470 535, 43 67-017 650, 10 65-109
INDO-1 Calcium 360 405, 10 65-072 486, 10 65-085
NADH/TAMRA ATPase 360 472, 30 67-013 572, 28 84-084

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Warnings
CAUTION: The optical input feeds light into the photomultiplier, which is an
extremely sensitive light measurement device. To avoid damage to the
photomultiplier, it is imperative that the probe be attached to the optical
input or that the input be capped when the SI-BF-100 is powered on.
PHOTOMULTIPLIERS ARE EXPENSIVE TO REPLACE.
Parts List
After unpacking, verify that there is no visible damage to the system. Verify that all
items are included:
(1) SI-BF-100 Biofluorometer
(1) 12V Power adapter
(1) Species-specific probe (for example, calcium or ATPase)
(1) M3301 Manual Manipulator
(1) M10 Magnetic Stand
(1) Instruction Manual
Unpacking
Upon receipt of this instrument, make a thorough inspection of the contents and
check for possible damage. Missing cartons or obvious damage to cartons should be
noted on the delivery receipt before signing. Concealed damage should be reported
at once to the carrier and an inspection requested. Please read the section entitled
“Claims and Returns” on page 27 of this manual. Please contact WPI Customer Service
if any parts are missing at 941.371.1003 or [e-mail].
Returns: Do not return any goods to WPI without obtaining prior approval (RMA
# required) and instructions from WPI’s Returns Department. Goods returned
(unauthorized) by collect freight may be refused. If a return shipment is necessary,
use the original container, if possible. If the original container is not available, use a
suitable substitute that is rigid and of adequate size. Wrap the instrument in paper
or plastic surrounded with at least 100mm (four inches) of shock absorbing material.
For further details, please read the section entitled “Claims and Returns” on page
27 of this manual.

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INSTRUMENT DESCRIPTION
The SI-BF-100 works with the SI-H muscle tester systems like the SI-MKB.
Calcium Probe
SI-MKB Platform (sold separately) SI-BF-100 Biofluorometer
Fig. 2—The probe is mounted in a micromanipulator and positioned in the muscle tester
cuvette in close proximity to the tissue sample.
Front Panel
Probe
Excitation
Port
Probe
Emission
Port 1
Probe
Emission
Port 2
BNC Outputs
Display Control Buttons
For Future Use
Port
or
t 1
ort
2
t
N
N
NC Out
ut
NC Ou
NC Ou
p
LED Module
Status Indicator
For Future U
s
Power Button
Fig. 3—The Biofluorometer control unit is rack mountable.
Probe Excitation Port–Connect the excitation fiber of the probe to this port. All the
LED modules are connected inside the chassis to this port using fiber optic cables
and a combiner. When the LEDs are turned on, the light travels through this port to
the probe tip.
Probe Emission Ports–Connect the emission fiber(s) of the probe to these ports.
Fluorescent emissions from the tissue travel from the probe tip to these ports and
into their respective photomultipliers. Some applications (like calcium concentration

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measurement) only require one photomultiplier. Others (like the ATPase
measurement) require both photomultipliers.
BNC Outputs: The analog outputs can be connected to a data acquisition system.
They provide a normalized (relative) output based on the LED intensity and the PMT
(photomultiplier) gain factor. The output depends on the system application. For
example, for calcium measurement:
• Output A–Normalized output of the two 340nm LED modules
• Output B–Normalized output of the 380nm LED module
• Output A/B–Ratio of Output A to Output B
NOTE: The display shows the actual ratio of the emissions measured from the two
frequencies of light. To calculate the ratio based on the A/B BNC voltage output, use
the formula:
Ratio = A/B Voltage x 2
Gain Adjustment Factor
Display–Press the Display button to toggle between the system parameters and the
live ratio measurements. Press the Setup button to enter the Configuration mode
and change the system setup.
Fig. 4—The display (left side) shows the actual ratio measurement. The six control buttons
(right side) let you setup the configuration parameters, turn the LEDs off and on, and
manipulate the display.
Control Buttons–There are six control buttons.
• Display–Press this button to toggle the display between parameters and
ratio measurement.
• Setup–Press this button to enter the setup mode and change parameters.
Adjustable parameters include sampling frequency, gain adjustment,
photomultiplier gain voltage and filter frequency.
• Module Config–Press this button to configure any of the LED modules.
From this menu, you can enable/disable a module, set the current output,
set the delay before it illuminates and set the length of the period that the
LED will illuminate.
• LED On/Off–Press this button to turn all the enabled LED modules on or
off. When the enabled modules are on, the LED light flashes. When you are

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modifying the LED configuration the LED light illuminates without flashing
so you can see the measurement of the LED output on the display.
• Up/Down–Use the buttons to modify configuration setting or parameters.
Power Button–Use this toggle to turn the unit power off or on.
NOTE: The USB connection is reserved for future development.
Back Panel
LED Module Monitor Port
L
Power Supply Connector
L
L
P
Fig. 5—The back panel has a power supply connector and a connection to monitor the on/
off status of the LED modules.
Power Supply Connector–Connect the 12V power adapter to this port.
LED Module Monitor Port–This port gives you access to each LED module to trigger
activity based on a wavelength that is firing. It uses a 5.0V TTL pulse with high
indicating that the LED is firing and low indicating that the LED is off. The unit can
hold up to seven LED modules. Three modules are included with the SI-BF-100 that
is configured for calcium measurement.
Setting Up the System
1. Mount the M10 magnetic stand on the back of the muscle tester platform.
2. Mount the M3301 micromanipulator on the M10 stand.
3. Install the probe in the micromanipulator (Fig. 6).
Fig. 6—(Left) The probe is installed in the micromanipulator.
Fig. 7—(Right) The probe tip is positioned between the tissue holders next to the tissue.

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4. Position the probe so that it fits into the muscle tester cuvette between the
tissue mounts (Fig. 7).
5. If the probe has two fiber optic cables connected to the head, then plug the fiber
optic cable labeled Excitation into the port on the controller labeled LED Optical
Output. Plug the other fiber optic cable (labeled Emission) into the port labeled
Optical Input A. Some probes have two emission fibers. Plug the fiber optic
cable labeled Emission A into port labeled Optical Input A. Plug the fiber optic
cable labeled Emission B into port labeled Optical Input B.
Excitation
Fiber
Emission
Fiber
Fig. 8—Press the SMA connectors into place and tighten the nut to ensure that stray light
does not enter the fiber connection.
CAUTION: The optical input feeds light into the photomultiplier, which is an
extremely sensitive light measurement device. To avoid damage to the
photomultiplier, it is imperative that the probe be attached to the optical
input or that the input be capped when the SI-BF-100 is powered on.
PHOTOMULTIPLIERS ARE EXPENSIVE TO REPLACE.
Changing the PMT Filters
Each photomultiplier has a filter on the front of the SI-BF-100 unit. The filter limits
the emission light that gets sent to the photomultiplier to a single bandwidth of the
spectrum. The filter can be easily swapped out or removed. The filters are inside
the filter holders located behind the SMA connectors labeled Optical Input A and
Optical Input B.
1. Turn off the power to the SI-BF-100 and unplug the instrument.
CAUTION: The optical input feeds light into the photomultiplier, which is an
extremely sensitive light measurement device. To avoid damage to the
photomultiplier, it is imperative that the probe be attached to the optical
input or that the input be capped when the SI-BF-100 is powered on.
PHOTOMULTIPLIERS ARE EXPENSIVE TO REPLACE.

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Filter Holder
Mounting Screws
S
S
Fig. 9—The filters are located inside the filter holders.
2. Use a Hex key to remove the two mounting screws that secure the filter holder
to the face of the SI-BF-100 (Fig. 10).
Fig. 10—(Left) Remove the two mounting screws.
Fig. 11—(Right) When viewed from the back side of the filter holder, you can see the filter.
3. Pull the filter holder straight off of the chassis (Fig. 11).
4. Use a small hex key to loosen the set screw holding the filter in place. (Fig. 12).
Fig. 12—(Left) Use a hex key to loosen the set screw holding the filter in place.
Fig. 13—(Right) The filter was removed from this filter holder.
5. Gently invert the filter holder onto a piece of paper so that the filter falls out.
6. Position the new filter in the filter holder, replace the cover and tighten the set
screw.
7. Line up the holes on the filter holder with the screw holes on the face of the
chassis. The indents on the bottom of the filter holder fit over the screws heads
that protrude from the face of the chassis.

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Fig. 14—Line up the screw holes as illustrated here.
8. Insert the screws and tighten them down to re-mount the filter holder on the
face of the Biofluorometer chassis.
Fig. 15—The filter holder has been reinstalled.
Adding an LED Module
Each SI-BF-100 is configured for its intended application. For example, the
intracellular calcium (Fura-2) measurement system uses three LED modules and
one photomultiplier. The Biofluorometer can be configured with up to seven LED
modules and two photomultipliers. You can add additional LED modules, if required.
1. Turn off the power to the SI-BF-100 and unplug the instrument. Then, use a
Phillips screw driver to remove the four screws on the top of the Biofluorometer
box.
2. Lift the lid and remove it to reveal the circuitry inside. The seven LED ports are
located in the front of the box. The are labeled 0–6 as shown in Fig. 16.

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SI-BF100 Front
PMT 1
PMT 2
LED 2LED 0 LED 4 LED 6
LED 1 LED 3 LED 5
SI-BF100 Circuit Board
LED
Module
Remove Screws
Fig. 16—(Left) The circuit board inside the Biofluorometer holds up to seven LED modules
and has two photomultipliers.
Fig. 17—(Right) Each LED module is secured to the circuit board by two screws located on
either side of it.
3. To remove an LED module:
Disconnect the BNC terminated end of the fiber optic cable that comes out of
the back end of the LED module.
Loosen the two screws on the sides of the LED module (Fig. 17).
Lift the LED module off the circuit board, being careful not to bend the fiber
optic cable.
4. To install an LED module:
Align the serial connection on the bottom of the LED module with the
appropriate LED port on the circuit board.
Press the LED module into place and secure it by tightening the two screws on
the sides of the LED module.
Connect the BNC-terminated end of the fiber optic cable to the proper emission
port.
Fig. 18—(Left) There are seven, 9-pin LED module ports on the circuit board.
Fig. 19—(Right) The LED module is installed on the board.

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Changing the Sample Averages of the Display
The sample average is shown on the display with the actual emissions ratio. The
sample average (AVG SAMPLE) represents the number of samples that are averaged
to arrive at the ratio measurement displayed. The system displays a running average
of the measurements. The larger the number, the slower the system is to respond to
changes.
NOTE: When you modify the sample average, the measurement algorithm changes.
The system will need a few minutes to settle after any changes are made.
NOTE: Making changes here does not affect the data from the BNC outputs.
To change the number of samples to average:
1. If the Ratio and Avg Samples are not shown on the display, press the Display
button to toggle the display.
Fig. 20—Press the Display key until the AVG SAMPLES is shown on the display.
2. Press the Up or Down button to change the AVG Samples value. The default
number of samples is 20.
Changing the Operating Mode
The Biofluorometer has two operating modes, single emission and dual emission.
Choose the operating mode based on your
application. The operating mode displays on the
bottom line of the Parameters Display. (See
“Parameters Display” on page 15.)
Fig. 21—(Right) This Parameters Display window shows
that the system is operating in Single Emission mode.
If you need to change the operating mode:
1. Press and hold the Up and Down buttons simultaneously to access the
operating mode menu (Fig. 22). “Operating Mode Menu” displays briefly on the
screen. Then, the active operating mode displays (Fig. 23 or Fig. 24).
Fig. 22—(Left) The operating mode menu splash screen displays briefly.
Fig. 23—(Center) This example shows a unit that is operating in single emission mode.
Fig. 24—(Right) This example shows a unit that is operating in dual emission mode.
2. Press the Up or Down button to change the operating mode. The operating
mode changes on the main Parameter Display screen.

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Fig. 25—Dual display on the bottom line of the Parameter Display screen when the unit is
operating in Dual Emission mode.
3. Press the Display button to return to the main display.
Configuring the System
Press the Setup button to enter the system configuration menu. This menu toggles
between the options that allow you to adjust the sampling frequency, gain adjust,
photomultiplier gain voltage and filter frequency.
Sampling Frequency
Gain Adjust
PMT1 Gain Voltage
PMT2 Gain Voltage
Filter Frequency
Fig. 26—Press the Setup button to cycle through the configuration parameters.
Modifying the Sampling Frequency
The sampling frequency determines the length of the sampling period. In essence, it
sets how quickly measurements are taken. The default is 1000Hz, and settings can
range from 1–1000Hz.
1. Press the Setup button. The current sampling frequency is shown on the display.
Fig. 27—The sampling frequency can be set from 1–1000Hz.
2. Press the Up or Down button to change the sampling frequency.
3. Press the Display button to save the configuration and return to the main
display.
Modifying the Gain Adjust
By default the gain factor is set to 1.000. At a gain of 1.000 the Output A/B records
half of the actual measurement of the ratio. The gain adjust can be set from 1.00 to
4.10. The display always shows the actual emission reading. However, if you use the
BNC outputs, you must compensate for the gain adjustment factor. The actual ratio
is twice the Output A/B reading divided by the gain factor.
(2 x Output)/Gain Adjust = Actual ratio

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1. Press the Setup button until the Gain Factor parameter displays. The current
gain factor is shown on the display.
Fig. 28—The gain adjust can be set from 1.00–4.10.
2. Press the Up or Down button to change the gain adjust.
3. Press the Display button to save the configuration and return to the main
display.
Modifying the Photomultiplier Gain Voltage
By default the gain voltage is set to 0.702V. This voltage can be set from 0.5–1.0V.
The photomultiplier front end optical gain increases as this voltage increases.
NOTE: PMT2 Gain Voltage modulates the gain factor for the second photomultipier
in the same manner.
1. Press the Setup button until the PMT1 Gain Voltage or PMT2 Gain Voltage
parameter displays. The current PMT1 gain voltage is shown on the display.
Fig. 29—The gain voltage can be set from 0.5–1.0V.
2. Press the Up or Down button to change the gain voltage.
3. Press the Display button to save the configuration and return to the main
display.
NOTE: The noise increases as the gain voltage increases.
Modifying the Filter Frequency
A programmable low pass filter is used at the photomultiplier output for noise
reduction. By default the filter frequency is set to 2800Hz. This is the frequency of
the photomultiplier video signal filter applied to the emission wavelength.
1. Press the Setup button until the Filter Frequency parameter displays. The
current filter frequency is shown on the display.
Fig. 30—The filter frequency can be set from 10kHz to 100Hz
2. Press the Up or Down button to change the filter frequency.
3. Press the Display button to save the configuration and return to the main
display.

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Setting up LED Modules
Each LED module can be individually configured. It can be enabled/disabled. You
can set the output current, determine when (in a period) the LED illuminates and
how much of the period that the LED remains lit. You can also see the combined
output of the all enabled LEDs.
1. Press the Module Config button to enter the module configuration menu. The
menu for Module 0 displays. To configure a different module, press the Module
Config button again until the desired module appears. Note two example
configurations are shown in the table below
Module Fura-2 Calcium Measurement ATPase Measurement
Module 0 340nm 370nm
Module 1 340nm 370nm
Module 2 380nm 370nm
Module 3 Not used Not used
Module 4 Not used Not used
Module 5 Not used Not used
Module 6 Not used Not used
Fig. 31—Press the Module Config button to reveal the module configuration menu.
2. Press the Setup button until the arrow displays next to the parameter you wish
to adjust.
• Current–Increasing the current boosts the power and the light output of the
LED module. The current can be adjusted in 1mA increments.
• Delay–The delay determines how far into a period before the LED module
turns on. In this example, LED module 0 illuminates after a 1% delay time.
• Width–The width is the percentage of the period that the LED module
remains lit. In this example, LED module 0 will remain on for 33% of the
period (Fig. 32).
• Enable–Enable allows the module to illuminate during a cycle. Disabled
modules will not illuminate. You can also see the enable/disable status of
an LED module on the main parameters display. (See “Parameters Display”
on page 15.)
NOTE: OUTPUT is a read-only parameter that shows the active output of all enabled
LED modules.

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33% 33% 32%
1%
Sampling Frequency
1000Hz
Module 0 On
Module 1 On Module 2 On Dark
Calcium Measurement
50% 49%
1%
Sampling Frequency
1000Hz
Modules 0, 1, and 2 On Dark
ATPase Measurement
Fig. 32—Two example configurations are shown here. The default setup for a calcium
(Fura-2) measurement unit has modules 0 and 1 (340nm) delay for 1% of the period and
illuminate for 33%. After another 1% delay, module 2 illuminates for 33% of the period.
The rest of the period is dark. In the ATPase setup, the modules illuminate for 50% of the
period and then are dark.
3. Press the Up or Down button to modify the setting.
4. Press the Display button to save the configuration and return to the main
display.
OPERATING INSTRUCTIONS
Changing the Display
Press the Display button to toggle between the two main displays:
• Parameters display
• Ratio/AVG Samples display
Fig. 33—(Left) Parameters display
Fig. 34—(Right) Ratio/Average Samples display
Parameters Display
The Parameters Display screen shows setup configuration parameters:
Module State–The unit can hold up to seven LED modules (Fig. 36), and they are
numbered 0–6. “E” indicates that a module is enabled. “D” indicates that the module
is disabled. In Fig. 33 the first three modules (0–2) are enabled and the last four
(3–6) are disabled. (See “Setting up LED Modules” on page 14.)
LED 0-2: Enabled
LED: 0 1 2 3 4 5 6
LED 3-6: Disabled
Fig. 35—The Module State shows which LED modules are enabled.
Our example display shows a typical calcium measurement system. Only three LED
modules are installed on that system. In this calcium measurement system, Module
0 and Module 1 emit light at 340nm wavelength, and Module 2 emits light at the
380nm wavelength. Modules 3–6 are not used.

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Fig. 36—The LED modules inside the Biofluorometer are arranged as shown here.
PMT Voltage–The photomultiplier voltage can be set between 0.5 and 1.0V. By
default it is set to 0.7V. (See “Modifying the Photomultiplier Gain Voltage” on page
13.)
Filter Cutoff–The output of the photomultiplier tends to be noisy, so it has a
filter. The cutoff for the filter can be adjusted. By default it is set to 2800Hz. (See
“Modifying the Filter Frequency” on page 13.)
Sample Frequency–The sample frequency determines how often measurements
are taken. You can set the frequency between 1 and 1000Hz. By default, the system
is set at 1000Hz. The system turns the 340nm LED modules on for a third of the
period, then it turns the 380nm LED module on for a third of the period, and finally
the last third of the period is dark. (See “Modifying the Sampling Frequency” on page
12.)
Ratio/Sample Averages Display
Typically, when an experiment is under way, you will observe the Ratio/Sample
Averages display screen.
Ratio–The ratio displayed is the actual ratio between the emission frequencies
measured.
Avg Samples–”Avg Samples” shows the number of samples that are averaged to
arrive at the Ratio measurement on the display. The system displays a running
average of the measurements. The larger the number, the slower the system is to
respond to changes. To change the number of samples to average, press the Up or
Down button. The default number is 20. The analog output is not affected by this
setting. It is for display purposes only.
Turning On/Off the Modules
Press the LED On/Off button to turn the LED modules on or off. When you turn the
LED modules on, the LED light begins to blink. The LED modules will cycle off an on
according to their programming. See “Setting up LED Modules” on page 14.
CAUTION: THE PHOTOMULTIPLIER ONLY WORKS WHEN THE LED
MODULES ARE ENABLED AND THE LEDS ARE ILLUMINATED. THERE-
FORE, THE OPTICAL INPUTS MUST BE CAPPED OR CONNECTED TO THE
PROBE AT ALL TIMES TO AVOID COSTLY DAMAGE OF THE PHOTOMULTIPLIERS.

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Configuration Parameters for Various Fluorophores
Parameters
Fluorophores
Fluo-4/Fura Red Indo-1 Fura-2 NADH/TAMRA
Primary LED Wavelength 470 360 340 360
Number of Primary LEDs 2222
Enabled Modules Primary LEDs
Current of Primary LEDs 25ma 25ma 25ma 25ma
%Delay of Primary LED 1111
% Width of Primary LEDs 50 50 33 50
Secondary LED Wavelength 380
Number of Secondary LEDs 1
Enabled Modules Secondary LEDs
Current of Secondary LEDs 25ma
%Delay of Secondary LEDs 1
% Width of Secondary LEDs 33
PMT1 Gain Voltage 0.600 0.600 0.600 0.600
PMT1 Filter 535 405 510 472
PMT2 Gain Voltage 0.600 0.600 0.600
PMT2 Filter 650 486 572
Sampling Frequency 1000 1000 1000 1000
Amplifier Gain 3.800 3.800 3.800 3.800
Filter Frequency 2800 2800 2800 2800
Filter Position 0000
MAINTENANCE
Monitoring the LED Output
Sometimes it is helpful to know the output of the individual LED modules. For
example, if you want to balance the power of the excitation frequencies, you might
want to know how much each module is putting out. This can be done by disabling
all the modules except the one(s) of interest. For example, in the intracellular calcium
(Fura-2) measurement system, Modules 0 and 1 emit 340nm light. If you disable
Module 2, you can determine the output of Modules 0 and 1. Likewise, if you
disable Modules 0 and 1, you can determine the output of Module 2. To balance the
frequencies, you can adjust the current sent to the individual modules.
1. Press the Module Config button to enter the module configuration menu. The
menu for Module 0 displays. To configure a different module, press the Module
Config button again until the desired module appears.
2. Enable/disable the desired modules. See “Setting up LED Modules” on page 14.
Fig. 37—The arrow appears next to the parameter that can be adjusted.