Harvard Bioscience CMA 470 User manual

8002778 Rev 1.0
CMA 470 Refrigerated Fraction Collector

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CMA 470 REFRIGERATED FRACTION COLLECTOR USER’S MANUAL
CONTENTS
1INTRODUCTION.................................................................................................... 3
2SAFETY................................................................................................................... 3
3UNPACKING & ASSEMBLY............................................................................... 4
3.1 Packing list ...................................................................................................... 4
3.2 Positioning the Unit........................................................................................ 4
3.3 Connections .................................................................................................... 4
3.4 Installation of accessories............................................................................. 4
4DESCRIPTION ....................................................................................................... 5
4.1 Control Panel................................................................................................... 5
4.2 Rear .................................................................................................................. 6
4.3 Power Switch................................................................................................... 6
4.4 Cassette Magazine.......................................................................................... 7
4.5 Vial cassettes .................................................................................................. 7
4.6 Cannula for Tubing......................................................................................... 9
4.7 Cannula for Septa ........................................................................................... 9
4.8 Single collect Assembly............................................................................... 10
4.9 Dual – Quadruple Collect Assembly........................................................... 11
4.10 Basic Functions ............................................................................................ 12
4.11 Safety Devices............................................................................................... 12
5OPERATING INSTRUCTIONS.......................................................................... 13
5.1 POWER on ..................................................................................................... 13
5.2 Set collection mode...................................................................................... 13
5.3 Set temperature............................................................................................. 14
5.4 Set first sample and number of samples ................................................... 14
5.5 Set fraction time............................................................................................ 15
5.6 Prepare for collection experiment .............................................................. 15
5.7 Connect the sample set up.......................................................................... 15
5.8 Start/Stop ....................................................................................................... 15
5.9 Save settings................................................................................................. 16
5.10 Reset settings............................................................................................... 16
5.11 Clean the system after use .......................................................................... 16
6ADVANCED OPERATION................................................................................. 16
7ACCESSORY KIT................................................................................................ 18
8OPTIONAL ACCESSORIES.............................................................................. 18
8.1 Vials ................................................................................................................ 18
8.2 Cassettes for collection into 2 mL vials..................................................... 18
8.3 Cables ............................................................................................................ 18
9MAINTENANCE & SERVICE ............................................................................ 18
9.1 Cleaning the Instrument............................................................................... 18
9.2 Storage........................................................................................................... 18
9.3 Warranty......................................................................................................... 19
9.4 Damaged shipments..................................................................................... 19
9.5 Service ........................................................................................................... 19
10 TROUBLESHOOTING.................................................................................... 20
11 TECHNICAL DATA ......................................................................................... 20
12 TEXT AND SYMBOL EXPLANATION ........................................................ 21
13 ORDER INFORMATION .................................................................................... 21
8002778 Rev 1.0

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Figure 1. CMA 470 Refrigerated Fraction Collector.
1 INTRODUCTION
The CMA 470 Refrigerated Fraction Collector is specially designed to collect
microliter volume fractions typical of microdialysis. It has thermoelectric
cooling down to +6 °C and the fractions can be collected in sealed vials. Both
of these are important considerations for the prevention of evaporation and
chemical degradation. It is possible to collect fractions as small as 1 µL at the
bottom of each vial. The capacity of the collector is 64 vials of 300 µL each or
40 vials of 2 mL each. When equipped with the quadruple cannula assembly,
the CMA 470 can collect from four sources into four vials simultaneously. With
the dual cannula assembly it can collect from two sources into two vials. There
is also an option to collect samples in open vials.
2 SAFETY
CMA 470 Refrigerated Fraction Collector is designed for laboratory use only,
e.g. animal experiments, analytical chemistry, etc, and is therefore not
equipped with the special safety functions that are necessary for use in
studies involving humans. In view of this, the following considerations should
always apply:
• The CMA 470 Refrigerated Fraction Collector should only be used
for its intended purpose, namely as a fraction collector for small
volumes in laboratory studies.
• The CMA 470 Refrigerated Fraction Collector should always be
used in accordance with the instructions in the User's Manual.
• The CMA 470 Refrigerated Fraction Collector should be used by
trained personnel who understand its proper use.
• If the CMA 470 Refrigerated Fraction Collector is sold or trans-
ferred, the new owner or user should be scientifically responsible and
have the capacity and expertise to use the fraction collector properly
and solely for its intended purpose.

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It is in the interests of your organization and of every person who is
responsible for the custody, operation, and maintenance of the CMA 470
Refrigerated Fraction Collector that the foregoing guidelines are complied with
at all times. In the event of an accident, failure to comply could result in legal
liability.
3 UNPACKING & ASSEMBLY
The CMA 470 Refrigerated Fraction Collector is delivered in a specially
designed box to protect the instrument against damage during transportation.
The reusable carton provides excellent protection should it be necessary to
transport the instrument or store it for a long period of time.
3.1 Packing list
The CMA 470 Refrigerated Fraction Collector, Ref No 800 2770 consists of:
- Refrigerated Fraction Collector equipped with Holder for Single Cannula
- External Power Supply
- EU Power Cable
- US Power Cable
- 4 Cassettes for Small Vials
- Holder for Quadruple Cannulae
- Holder for Dual Cannulae
- Cannula for Tubing 4 pcs
- Cannula for Septa 4 pcs
- User's Manual
After unpacking the instrument, check the contents against the above packing
list to ensure that the shipment is complete. Inspect all items for damage. Any
damage or missing parts should be reported immediately to your local supplier
or CMA/Microdialysis AB. Remove the transport security material.
3.2 Positioning the Unit
The CMA 470 Refrigerated Fraction Collector can stand directly on a
laboratory workbench or any other flat stabile, vibration-free surface adjacent
to a grounded wall socket (required).
NOTE: Make sure the drain tubing is not kinked under the instrument so that
the moisture from the cassette magazine can be drained. Place the outlet end
on the same level, or lower, as the bottom of the instrument.
3.3 Connections
For connection to the mains, use the External Power Supply (12 VDC, 55 W,
included).
Please note the following when connecting other CMA instruments to
your CMA 470 Refrigerated Fraction Collector:
Before plugging the Fraction Collector into the socket, check that the input
voltage and frequency correspond to the information given on the nameplate
on the power supply for the instrument.
3.4 Installation of accessories
For the installation of Cassettes, Holder for Cannulae and Cannulae refer to
Sections, 4.1 - 4.10 and 6.

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4 DESCRIPTION
The CMA 470 Refrigerated Fraction Collector consists of a temperature-
controlled cassette magazine (+6 °C to room temperature) and a control
panel. Vial storage is divided into four cassettes. When configured for
collection from a single source, the cannula can be maneuverable to one of
two collection positions. When equipped with the Holder for Dual Cannulae
two samples can be collected simultaneously and with the Holder for
Quadruple Cannulae three to four samples can be collected simultaneously.
Collection can be into either open or sealed vials.
The CMA 470 Refrigerated Fraction Collector is delivered complete with
power cables and external power supply. Once the plug has been connected
to a wall socket, the instrument is ready for use.
4.1 Control Panel
1 2 3 4 5 6 7 8
Figure 4.1 Control panel
Locate the following items on the control panel of the CMA 470 Refrigerated
Fraction Collector. See figure 4.1
Item
Description
1. Display
window
Indicates all basic settings and status:
Status
Mode
Current samples, fraction time, total time and
temperature.
Cooling activity
2. Menu
Advanced settings.
3. Reset
Push button to reset all running parameters.
4. Cassette
Rotates the magazine clockwise to the first
position in the next cassette.
5. Step
Move to next collecting position.
6. Start/Stop
Push button to start/stop the fraction collector.
7. Selection
Knob
For selection of mode, number of samples,
fraction time and temperature.
Turn to select, press to activate and accept.
Clockwise to increase or move up, counter
clockwise to decrease or move down. Cursor is
blinking when active.
8. Green LED
Green LED on – collection is running.

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4.2 Rear
1 2 3 4
Figure 4.2 Rear
The following are located on the rear of the CMA 470 Refrigerated Fraction
Collector, see Figure 4.2
Item
Description
1. USB
Socket for USB connection to PC or other
device with USB host port.
2. RS232
Socket for RS232 interface for connection to
PC or other device with serial port.
3. Digital I/O
Socket for digital Input/Output for external
equipment such as CMA 400 or other
instruments.
4. Power
Socket for 12 VDC inlet. Connection for power
supply.
4.3 Power Switch
1
Figure 4.3 Power Switch
Power Switch on the right side of the panel.
Item
Description
1. Power Switch
Switch the power on. The display illuminates.

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4.4 Cassette Magazine
1 2 3 4 5 6 7
Figure 4.4.1 Cassette magazine, closed lid Figure 4.4.2 Cassette magazine, open lid
Locate the following items for the cassette magazine of the CMA 470
Refrigerated Fraction Collector. See Figures 4.4.1 and 4.4.2
Item
Description
1. Lid
Lid to keep the cooling and to protect from light.
2. Lid handle
To open the lid.
3. Cassette
Removable cassettes, one of four.
4. Cassette
position
Numbering of cassettes
5.Supporting arm
for cannula
holder
To mount the holder for single or quadruple
cannulae on
6. Cannula for
Septa
For use when collecting in vials with Teflon
septa
7. Holder for
Single Cannula
For use in single collecting mode with cannula
for tubing or for septa.
4.5 Vial cassettes
Two options are available, 64 x 300 µl vials (H 35,OD 5 mm), or 40 x 2 ml vials
(H 35, OD 11 mm).
All four cassettes in the magazine must be the same model. Figures 4.5.1-2
show how to number fractions when using different cassettes and different
numbers of cannulae.
The collector is equipped with cassettes for small vials.

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Step sequences for small vials:
Figure 4.5.1 Numbering of the cassette positions for single, dual and
quadruple cannulae
NOTE: When three cannulae are in use, numbering is as for four cannulae.
Every fourth position will be empty.
Step sequences for large vials:
Figure 4.5.2 Numbering of the cassette positions for single and dual cannulae
NOTE: When collecting into large vials up to 2 cannulae can be used
simultaneously.

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To change cassettes, lift them by the knob and replace with the others, one at
the time. Press the cassette button to be able to release all.
Figure 4.5.3 Removal of cassette.
Warning: Risk for instrument damage.
Only use vial cassettes for CMA 470 Collector.
These cassettes are equipped with blue lift handles.
Cassettes belonging to CMA/170 Collector can
not be used, they will damage the instrument.
Contact CMA representative for more information.
4.6 Cannula for Tubing
To collect from tubing into the bottom of an open vial, use this option that
consists of a cannula and a plastic knob. The tubing from the probe is fed
through the supporting cannula until it touches the bottom of the collecting vial.
Secure the tubing in place by lifting the plastic knob.
NOTE: Make sure the tubing is not pushed so far into the vial that the end
bends.
4.7 Cannula for Septa
To collect into vials with Teflon septa, use this option that consists of a
beveled piercing cannula for the penetration of the septa.
Connect the tubing to the inlet of the cannula by the use of a Tubing Adapter.
NOTE: Use Teflon septa only.

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4.8 Single collect Assembly
Fig. 4.8.1 Setting up
Holder for Single
Cannula
Fig. 4.8.2 Setting up
Single Cannula for
Tubing
Fig. 4.8.3 Setting up Single
Cannula for Septa
4.8.1 Place the Holder for Single Cannula (1) so the tap (2) fit into the loop (3)
in the right side slot (4) on the Supporting Arm. Secure the holder with the two
accompanying screws (5). See Figure 4.8.1
4.8.2 Collecting in vials with tubing:
Insert the Cannula for Tubing in the Multi Holder for Cannula (6), adjust the
upper end of the metal cannula to the same level as the top of the Plastic
Locker Knob (7) when it is mounted in the Multi Holder for Cannula. Secure
the cannula with the screw in the Multi Holder for Cannula (8). See Figure
4.8.2
4.8.3 Collecting in vials with septa:
Place a waste vial in position 1 in the cassette and move it to the collection
position. Insert the Cannula for Septa (9) in the Multi Holder for Cannula (6).
Adjust the lower end of the cannula into the bottom of the vial (10). Secure
the cannula with the screw in the Multi Holder for Cannula (8). See Figure
4.8.3
4.8.4 To remove the Holder for Single Cannula, untight the two screws, turn
the Multi Holder for cannula so the opening of the locking washer (11), seen
under the supporting arm, is facing you. Lift up the Holder for Single Cannula
from the Supporting Arm. See Figure 4.8.1
2
3
4
5
5
6
7
8
6
8
7
9
10
11
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4.9 Dual and Quadruple Collect Assembly
Fig. 4.9.1 Setting up
Holder Quadruple
Cannulae
Fig. 4.9.2 Setting up
Quadruple Cannulae for
Tubing
Fig. 4.9.3 Setting up Quadruple
Cannulae for Septa
4.9.1 Place the Holder for Dual or Quadruple Cannulae (1) on the Supporting
Arm so that the Multi Holders for cannula (2) fit into the left slot (3) and the two
holes (3). Secure the holder with the two accompanying screws (4). See
Figure 4.9.1
4.9.2 Collecting in vials with tubing:
Insert the Cannulae for Tubing in the Multi Holders for Cannula (2), adjust the
upper ends of the metal cannulae to the same level as the top of the Plastic
Locker Knobs (5) when they are mounted in the Multi Holders for Cannula.
Secure the cannulae with the screws in the Multi Holders for Cannula (6). See
Figure 4.9.2
4.9.3 Collecting in vials with septa:
Place waste vials in the cassette and move them to the collection position.
Insert the Cannulae for Septa (7) in the Multi Holders for Cannula (2). Adjust
the lower ends of the cannulae into the bottom of the vials (8). Secure the
cannulae with the screws in the Multi Holders for Cannula (6). See Figure
4.9.3
4.9.4 To remove the Holder for Dual or Quadruple Cannulae, untight the two
screws, turn the Multi Holders for cannula (2) so the opening of the locking
washers (9), seen under the supporting arm, are facing you. Lift up the Holder
for Dual or Quadruple Cannulae from the Supporting Arm. See Figure 4.9.1
WARNING: Risk for cannulae damage.
When changing from quadruple mode to single or dual mode, always
change from the Holder for Quadruple Cannulae to the proper one
needed for the application.
Do not excessive tight the screw (6) to not damage the cannula
lumen.
A too loose screw (6) can’t hold the cannula in position.
1
2
5
3
3
5
4
4
2
6
7
8
5
6
2
6
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4.10 Basic Functions
• The vial cassettes rotate clockwise for positioning and sample collecting.
• The Holder for Single Cannula maneuver the cannula into one of two
collection positions, an inner and outer position.
• The Holders for Dual and Quadruple cannulae have fixed positions for 2
(dual), 3 or four cannulae (quadruple).
• The vial cassettes are maneuvered to the low position for the change of
collecting vials.
• The vial cassettes are in upper position for collecting sample(s).
• When the vial cassettes are lifted to the high position, fractions are
collected directly in the bottom of the vials. In this way, a very high
degree of precision is achieved and fractions smaller than a falling
droplet can be collected in a reproducible manner. When fraction are
collected in vials fitted with a septum, some of the liquid may be lost
against the septum upon withdrawal of the piercing needle such that the
quantity collected in each vial may vary depending on the rate of flow.
Change of fractions is controlled by time intervals, or by external signals.
• The cassette magazine is temperature-controlled by means of a
thermoelectric cooler with a fan.
4.11 Safety Devices
Keep finger and other objects away from moving parts in the cooling
compartment. Be careful when handling cannulae and collecting accessories.
WARNING:
Moving parts can cause injury and/or damage the collector.

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5 OPERATING INSTRUCTIONS
The CMA 470 Refrigerated Fraction Collector is easy to use. It is advisable to
read this user manual carefully before starting to use the instrument.
The CMA 470 Refrigerated Fraction Collector CMA can collect fractions from
up to four sources into four vials simultaneously. The cannulae can be
individually fitted and removed.
Prepare the instrument for the experiment (cassettes and cannulae).
See 4.5 - 4.9 for setting up the system.
5.1 POWER on
Connect to the external power supply. Switch POWER on. The display
illuminates. (Fig. 4.1 – 4.3)
5.2 Set collection mode
Available Modes: Single, Dual and Quadruple mode.
NOTE: Use Quadruple mode for triple cannula collection.
Turn the Selection Knob to move the cursor to the upper row.
Press the Selection Knob to activate the selection.
Turn the Selection Knob to set the desired mode.
Press the Selection Knob to acknowledge and lock the setting.
Figure 5.2 Setting collection mode

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5.3 Set temperature
NOTE: Turn on the cooling approximately 30 minutes before use.
The cooling capacity is -15 °C from environmental temperature or better.
Turn the Selection Knob to move the cursor to the lowest row.
Press the Selection Knob to activate the temperature setting.
Turn the Selection Knob to set the desired temperature.
Press the Selection Knob to acknowledge. The curser moves to the data field
for cooling on/off selection.
Turn the Selection Knob to select cooling on/off.
Press the Selection Knob to acknowledge and lock the setting.
Figure 5.3 Setting temperature
5.4 Set first sample and number of samples
Sample collection starts one position after the current position (waste position)
in the cassette magazine.
NOTE: First collecting fraction will be the position after the cannula location at
the start.
Turn the Selection Knob to move the cursor to the second row.
Press the Selection Knob to the position for number of samples.
Turn the Selection Knob to select the desired number.
Press the Selection Knob to acknowledge and lock the setting.
Figure 5.4 Setting number of samples

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5.5 Set fraction time
Turn the Selection Knob to move the cursor to the third row.
Press the Selection Knob to activate the position for the number of hours.
Turn the Selection Knob to set the desired value.
Press the Selection Knob to acknowledge. The curser moves to the data field
to the position for the number of minutes.
Turn the Selection Knob to set the desired value.
Press the Selection Knob to acknowledge. The curser moves to the data field
to the position for the number of seconds.
Turn the Selection Knob to select the desired value.
Press the Selection Knob to acknowledge and lock the settings.
Figure 5.5 Setting fraction time
5.6 Prepare for collection experiment
Load the cassettes with collection vials. Position waste vials at the first
position. Press Cassette and Step buttons to position the waste vials at
cannula/e position.
See Figures 4.5.1 and 4.5.2 for numbering of vials.
5.7 Connect the sample set up
When collecting straight from the tubing into the bottom of the collection vial,
feed the tubing down to the bottom of the waste vial and secure it by lifting up
the plastic knob.
See Figure 4.8.2 and 4.9.2
When collecting into vials with septa, connect the tubing to the collection
cannula with a Tubing Adapter.
See Figure 4.8.3 and 4.9.3
5.8 Start/Stop
Press "Start/Stop" to begin operation. The collector will move one step and
start the time mode fraction collection. The green LED will illuminate.
See Figure 4.1

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5.9 Save settings
All settings will be stored when the instrument is turned off. When restarting
the instrument the last settings will be activated.
5.10 Reset settings
When in ‘Stopped mode’ press the ‘Reset’ button to restart collection, using
the last saved settings.
In ‘Running mode’, use the reset button to restart the time for the ongoing
fraction. By pressing the Step button the collector will move to the following
fraction.
5.11 Clean the system after use
When finished with collection using the piercing Cannulae for Septa, flush
them with deionized water and then 70% ethanol. Use vials for waste.
Dry the magazine with a soft cloth.
6 ADVANCED OPERATION
With the Menu button a set of commands and service functions can be
activated.
For the user, only the Manual commands are useful.
When setting up the CMA 470 for a new sample method,
single/dual/quadruple mode or for the set up of collection for tubing or
cannula, manual commands is used to control sampling sequences.
When the Manual function is activated cassettes can be moved Up or Down
and the Multi Holder can be moved from the Outer to Inner vial row and back
(Single mode) with commands from the Menu.
Press the front panel buttons Cassette, Step and the Menu/Manual commands
Up/Down and In/Out when setting up the CMA 470 as described in chapter 4.8
and 4.9.
Press the Menu button.
Turn the Selection Knob to move the cursor to the second row.
Press the Selection Knob to activate the Manual functions.
Turn the Selection Knob to select the desired function.
Press the Selection Knob to acknowledge and execute selected function. To
execute the function a second time, press the Selection Knob.
Turn the Selection Knob to select a new function.
Press the Selection Knob to acknowledge and execute selected function.
Press the Menu button to exit.

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Figure 6.1 Menu options
Figure 6.2 Menu Manual operations
The menu functions Set-Up and Info are only for maintenance and service
purpose. Information can be found in the technical manual.

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7 ACCESSORY KIT
The CMA 470 Refrigerated Fraction Collector accessory kit includes:
- Holder for Dual Cannulae
- Holder for Quadruple Cannulae
- Cannula for Tubing 4 pcs
- Cannula for Septa 4 pcs
NOTE: At delivery the fraction collector is equipped with cassettes for small
vials and Holder for Single Cannula.
8 OPTIONAL ACCESSORIES
8.1 Vials
The fraction collector can be run with 300 uL plastic or glass vials, OD: 6 mm,
height: 35 mm (max) or 2mL glass vials OD: 12 mm, height: 35 mm (max)
Crimp Caps with Teflon membrane( non-resealing) can be used.
8.2 Cassettes for collection into 2 mL vials
When collecting into large vials these cassettes must be used. Large vial
cassettes must be placed in all four positions in the magazine. A maximum of
two collection cannulae can be used with this option.
8.3 Cables
CMA RS232 PC/RJ45 Cable for computer control of CMA 470.
USB cable type A-B for computer control of CMA 470.
9 MAINTENANCE & SERVICE
The CMA 470 Refrigerated Fraction Collector does not require periodic
lubrication or changes of any parts.
9.1 Cleaning the Instrument
Keep your CMA 470 Refrigerated Fraction Collector clean. Wipe off
any spillage using a soft cloth with mild detergent or 70% alcohol.
Periodic wipe off the cooling compartment.
9.2 Storage
If the CMA 470 Refrigerated Fraction Collector will not be used for a significant
length of time:
Disconnect the mains supply
Clean the instrument
Wipe off the cooling compartment.
Store the instrument away from dust and moisture

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9.3 Warranty
CMA/Microdialysis AB (called CMA) guarantees all components of the CMA
470 Refrigerated Fraction Collector to be free from defects of material and
workmanship for a period of one year after initial purchase.
For warranty service or repair, all CMA products must be returned to CMA or
to an authorized representative.
The foregoing warranty shall not apply to defects resulting from improper or
inadequate maintenance by the Owner, Owner-supplied software or
interfacing, unauthorized modification or misuse, operation outside of the
environmental specifications for the product, or improper site preparation or
maintenance.
For any product expressly covered under this warranty, CMA is liable only to
the extent of replacement of the defective items. CMA shall not be liable for
any personal injury, property damage, or consequential damages of any kind
whatsoever. The foregoing warranty is in lieu of all other warranties of
merchantability and fitness for a particular purpose.
9.4 Damaged shipments
Breakages of any part of this instrument during shipping should be reported
immediately to CMA Customer Service or an authorized representative. You
must retain the original packing box and contents for inspection by the freight
handler. CMA will replace any new instrument damaged in shipping with an
identical product as soon as possible after the claim filing date. Claims not
filed within 30 days after the shipping date will be invalid. Do not return
damaged goods to CMA without first contacting Customer Service.
9.5 Service
CMA and CMA distributors have skilled service staff to solve your technical
problems if an equipment-oriented problem should arise. For further details,
call/email/fax first your local CMA distributor and secondly direct to CMA.
Harvard Apparatus
84 October Hill Road
Holliston, MA 01746 USA
Phone Orders: 800-232-2380 • Fax: [phone]
E-mail: [e-mail] • www.harvardapparatus.com