CoolLED pE-340fura User manual

DOC-034 Iss 3 1
User Manual
pE-340fura

DOC-034 Iss 3 2
Table of Contents
1. Introduction .................................................................................................................... 3
2. Safety Precautions........................................................................................................... 4
3. Getting Started –System Components........................................................................... 6
4. Installation and setup...................................................................................................... 7
5. Configuration of the Light Source ................................................................................... 9
6. Operation - Manual Control.......................................................................................... 10
7. Remote Operation –TTL ............................................................................................... 13
8. Remote Operation –USB .............................................................................................. 18
9. Optical setup ................................................................................................................. 20
10. Additional Filtering Capability....................................................................................... 23
11. Excitation Filter Specifications ...................................................................................... 24
12. Settings / Additional Information ................................................................................. 26
13. Routine Care and Maintenance .................................................................................... 28
14. Fitting the pE-340fura Illumination System to a different microscope........................... 28
15. Product specifications ................................................................................................... 30
16. Product options and order codes.................................................................................. 30
17. Warranty and Repairs ................................................................................................... 31
18. Compliance.................................................................................................................... 31
19. Contact Details .............................................................................................................. 32

DOC-034 Iss 3 3
1. Introduction
CoolLED’s pE-340fura Illumination System has been designed to meet the
requirements of users carrying out Fura-2 AM ratiometric calcium imaging.
The pE-340fura offers an Illumination System that is both safer and offers a
faster level of control than traditional bulb-based solutions. The three
controllable channels offer 340nm, 380nm and a broad ‘white’ (420-700nm).
With a comprehensive range of microscope adaptors, the pE-340fura can be
fitted to most current and older compound research grade microscopes. The
result is a safe, convenient illumination system which will last for many years
without any additional operating costs.
This User Manual should give you all the information required to install and
operate your new illumination system.
Additional information can be found on our website at www.coolled.com

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2. Safety Precautions
While LEDs are a much safer illumination system than the mercury and
metal halide lamps that they replace in microscopy applications, precautions
should still be taken with this product.
When operating or maintaining this product, please observe the following
safety precautions at all times. Failure to do so may result in personal injury
or damage to other items.
2.1.
UV light may be emitted from this product depending on the
version/wavelength selected. Avoid eye and skin exposure. Never look
directly into the light output beam from the Light Source or accessories. The
emissions could damage the cornea and retina of the eye if the light is
observed directly.
2.2.
Always ensure that the Light Source is securely attached to the microscope
(either directly or with a light guide and collimator, depending on the
version) prior to turning on the power. This will minimise the risk of injury
and damage.
2.3.
If for any reason the Light Source is to be operated when not attached to a
microscope, all personnel should wear eye shielding and clothing to protect
the exposed skin.
2.4.
Disconnecting the mains supply is achieved by unplugging the power cord
from the power supply block or the Light Source. Only plug in the power
cable, once the Light Source is attached to the microscope.
2.5.
There are no serviceable parts within the Light Source. Removing any of the
screws and covers will result in the safety of the Light Source being impaired.
The DC power supply unit should be inspected periodically throughout the
lifetime of the system.
2.6.
Any electronic equipment connected to this product must comply with the
requirements of EN/IEC 60950.

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2.7.
To clean the exterior of the Light Source, use a slightly dampened cloth with
a simple water/detergent solution only. Avoid the optical surfaces and
lenses. Cleaning of optics should only be carried out using optical wipes and
fluids. Please note that the DC power supply unit should be isolated prior to
cleaning.
2.8.
This product conforms to the requirements of the Safety Standards as
follows:
EN/IEC 61010-1:2010 Safety Requirements for Electrical Equipment
for Measurement, Control and Laboratory use.
EN62471:2008 Photo-biological Safety of Lamps and Lamp
Systems/Guidance on manufacturing
requirements relating to non-laser optical
radiation safety. Risk Group 3.
All warnings may not be applicable depending on the version/wavelength being used.

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3. Getting Started –System Components
A typical CoolLED pE-340fura Illumination System is supplied with the
following components:
1. LED Light Source.
2. Manual Control Pod.
3. Microscope adaptor for specific microscope model (Direct fit only).
4. DC Power Supply Type GST120A12-R7B.
5. IEC Power Cable (not shown).
6. User Guide (not shown).
If any components are missing or appear damaged, please contact CoolLED
immediately.

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4. Installation and setup
4.1.
Carefully unpack the components from the shipping cartons.
4.2.
Insert the Control Pod cable into the LED Light Source using the red dots as a
guide for orientation of the plug.
4.3.
Connect the power connector from the DC power supply as shown. Ensure
that the DC power supply is the one supplied with the product. Using non-
CoolLED power supplies may damage the Light Source and will invalidate the
warranty. At this stage do not connect the mains power lead to the DC
power supply.

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4.4.
Attach the LED Light Source to the epi-fluorescence port on your
microscope. Your pE-340fura Light Source will have been supplied with a
compatible fitting for the microscope you specified at order (if direct fit
version). Attach the Light Source ensuring that it is secure and true/flush
with the microscope.
4.5.
Ensure that there is free airflow around the LED Light Source so that the
cooling system is not impaired. A gap of 200mm on either side is sufficient.
The diagram shows the Light Source in the preferred orientation. However it
may be set with the cables at the top or at either side.
4.6.
With the LED Light Source now attached to the microscope it is safe to
connect the mains power. Connect the mains lead supplied to a convenient
socket, plug in the IEC connector into the DC power supply and switch the
power on at the socket.

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5. Configuration of the Light Source
5.1.
5.2.
The pE-340fura has independent circuits giving the user control of the three
main peaks of emissions. These are referred to as 340 (nm), 380 (nm) and
3WT (white illumination).

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6. Operation - Manual Control
6.1.1.
Manual Control Pod Operation on/off.
The pE-340fura is easily controlled from the manual Control Pod. LEDs are
switched on and off by pressing the ‘on/off’ button.
6.1.2.
At start-up the Light Source will revert to the same settings that were set
when it was last powered down. New Light Sources are supplied with the
settings as shown.
6.1.3.
To switch on LEDs press ‘on/off’ once.

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6.1.4.
To switch off the LEDs, press the ‘on/off’ once again.
6.1.5.
Intensity Control.
The Control Pod enables the user to control the intensity of the LEDs that
are exciting different stains. This helps to balance the emissions so that one
stain does not dominate another.
6.1.6.
Reduce intensity of one channel by pressing the down intensity button.

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6.1.7.
Individual channels can be switched off (de-selected) by pressing the ‘select’
button. Light is then only generated where it is required to excite the stains
in use. This has many attractive benefits with improvements in contrast, cell
viability and savings in energy.
6.1.8.
Switch off selected channels by pressing the ‘on/off’ button.
6.1.9.
Switch selected channels back on by pressing ‘on/off’ button again.

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7. Remote Operation –TTL
7.1. Global triggering
7.1.1.
The pE-340fura has a BNC socket on the rear of the Light Source which allows
global control of the Illumination System.
The TTL signal controls the on/off function of the Light Source. A TTL ‘high’
will cause the LEDs to be on, independent of the state of the on/off button.
Only those bands which have been manually selected on the Control Pod
(shown by a shaded intensity bar on the Control Pod display) will be
switched by the TTL signal. The intensities of the selected bands are
manually set on the Control Pod.
7.2. Individual channel triggering
7.2.1.
In addition to the global TTL control available, the pE-340fura also has three
additional BNC sockets that allow individual TTL channel control of the
Illumination System.
Global BNC
socket
Channel 1
Channel 2
Channel 3

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7.2.2.
The TTL signal controls the on/off function of the Light Source. A TTL ‘high’
will cause the LEDs to be on. The channel controls shall trigger the
corresponding channel regardless of its on/off state or whether it has been
selected using the Control Pod. The intensities of the selected bands are
manually set on the Control Pod.
7.3. Sequence Runner
7.3.1.
The pE-340fura allows the Illumination System to be controlled using the
Sequence Runner mode. The Sequence Runner allows sequential triggering
of multiple channels using a single TTL signal attached to the global BNC
socket.
7.3.2.
The Sequence Runner mode is accessed by a short duration press of the
mode button on the Control Pod.
Global TTL (used for
Sequence Runner mode)

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7.3.3.
7.3.4.
Pressing the channel select button allows you to either deselect a channel or
change the order that it triggers in the sequence.
ON/OFF state of the
Illumination System.
Installed channels
Order in triggering sequence
Channel intensity
(0-100%)

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7.3.5.
Pressing the + & - buttons on the Control Pod will allow the light intensity of
the corresponding channel to be increased or decreased.
7.3.6.
The sequence will not begin until the ON/OFF button on the Control Pod is
pressed.
In this example channel 2 will pulse at 100% intensity, channel 3 will pulse at
65% and then channel 1will pulse at 100%. This Sequence will continue until
the ON/OFF button is pressed again to stop the sequence.
7.3.7.
Whilst a sequence is running the ability to change the triggering order, select
or deselect a channel and the mode button are disabled. The BNC sockets
responsible for Individual channel triggering are also disabled whilst in
Sequence Runner mode to avoid any conflicts.

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7.3.8.
An example of the TTL signal is shown below with labels showing the effect
on the light output during the sequence.
7.4. TTL triggering information
7.4.1.
The TTL input circuit has been designed to maximise the switching speed of
the LEDs to give the user precise control of the excitation light reaching the
sample.
1st channel
of sequence
2nd channel
of sequence
3rd channel
of sequence
1st channel
of sequence
This diagram shows the worst case triggering speeds when triggered at 100% intensity. There will be slight
differences in speed between channels and at different intensities.

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7.4.2.
With fast repetitive switching, the Control Pod display will not be able to
respond at the same speed. This can occasionally result in the Control Pod
displaying the incorrect ON/OFF state. If this happens, simply press the
ON/OFF button to reset the state on the display.
8. Remote Operation –USB
8.1.
For a remote control using software connection between the host computer
and the Illumination system, a USB interface is used. The Light Source has a
Type’B’ connector socket located adjacent to the Control Pod socket.
8.2.
Connect the Light Source to your computer using a USB cable. As with all
USB remotely controlled devices, it will be necessary to set up the driver
files on your system to allow the pE-340fura to be recognised.
8.3.
When you first plug your CoolLED system into your PC with the USB cable,
Windows will ask for a driver file unless one has already been installed. You
should point Windows to the file available from CoolLED.
USB

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8.4.
If you do not have the driver file you can download this from the following
page on the CoolLED website:
http://www.coolled.com/product-detail/imaging-software/
8.5.
Click on the link at the bottom of the Overview section titled ‘CoolLED
pE_Driver’. Download then unzip before pointing Windows to this file.
8.6.
Once the CoolLED device has been successfully installed into Windows you
should look at the Virtual COM ports assigned by going into Device
Manager. Look within Ports (COM & LPT).
In this example the Illumination System has been assigned two COM ports,
COM3 and COM4. You may need this information to connect to the Light
Source from your software control package. Either COM port may be used
for control. Two COM ports have been assigned to allow for diagnostics to
take place in parallel with communication and also allow for dual
communication should it ever be desired.
8.7.
The majority of microscopy imaging software systems have integrated the
pE-340fura into their packages. If you are developing your own software, a
Software Development Kit (SDK) is available giving the full instruction sets
necessary. Contact supp[email protected] and request access to this
information.

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9. Optical setup
9.1. Direct fit version
9.1.1.
The pE-340fura has been designed to work on the majority of fluorescence
microscopes, both new and old. As would be expected, there is some
variation in the optical path and elements within every microscope. In order
to accommodate these variations, the pE-340fura is supplied with a small
adjustment which allows the user to optimise the performance of the
illumination system when it is first fitted. This is a one-time adjustment. No
further adjustment will be required during the life of the product unless
changes are made to the microscope or the illumination system is fitted to a
different microscope.
9.1.2.
To make the adjustment, set up a typical sample on the microscope that
gives an image over the whole field of view. Loosen the thumbscrew and
slide the post back and forth until you achieve the maximum brightness
with an even field of view. Tighten the thumbscrew to prevent the setting
from changing.
Pull post
back and
forth to
make
adjustments
Thumbscrew