USER MANUAL
Flexcell
®
FX-5000
™
Compression System
06-20-17
Rev 16.0
Flexcell International Corporation 2730 Tucker Street, Suite 200 Burlington, NC 27215
Culturing Cells in a Mechanically Active Environment™
800-728-3714 (919) 732-1591 FAX: (919) 732-5196 www.flexcellint.com
COPYRIGHT © 2009 FLEXCELL
INTERNATIONAL CORPORATION
FLEXCELL
®
INTERNATIONAL CORPORATION
TABLE OF CONTENTS
One Year Limited Parts Warranty ................................................................................................ ii
Dell Computer Warranty Information ......................................................................................... iii
Federal Communications Commission Radio Frequency Interference Statement ...................... iv
FX-5000™ Quick Start Instructions .............................................................................................. v
1. Flexcell® FX-5000™ Compression System Components .................................... 1
2. Setup of the Flexcell
2.1 Packing List ...................................................................................................................... 1
2.2. Location of the FX-5000™ Compression System ............................................................ 1
2.3 Setup of the Computer and FX5K™ Compression FlexLink® .......................................... 2
2.4 Incubator Access Ports ...................................................................................................... 3
2.5 Sample Preparation and Setup of BioPress
Assembly........................................................................................................................... 4
2.6 Connecting the System Positive Pressure Lines ............................................................... 6
2.7 FlexSoft
2.8 LCD Display on the FX5K
2.9 Using Additional FlexLinks
®
FX-5000™ Software and System Power-up ..................................................... 7
®
FX-5000™ Compression System ...................................... 1
™
Culture Plates and Baseplate Clamping
™
Compression FlexLink® ..................................................... 7
®
............................................................................................ 8
3. Operation Instructions .......................................................................................... 9
3.1 Starting the FX-5000™ Program ....................................................................................... 9
3.2. Using the FX-5000™ Tutorial .......................................................................................... 9
3.3 Main Display Window ...................................................................................................... 9
3.4. Configuring Users .......................................................................................................... 11
3.5 Creating Regimens .......................................................................................................... 11
3.6 Creating a Regimen with a Custom Waveform Shape ................................................... 13
3.7 Assigning Regimens ....................................................................................................... 14
3.8 Starting Regimens ........................................................................................................... 14
3.9 Simulating Regimens ...................................................................................................... 14
3.10 Pausing Regimens ......................................................................................................... 15
3.11 Resuming Regimens ..................................................................................................... 15
3.12 Stopping Regimens ....................................................................................................... 15
3.13 Determine the Progress of the Regimen ....................................................................... 15
3.14 Resetting the FlexLink® ................................................................................................ 15
3.15 Using the Pressure/Force Conversion Calculator ......................................................... 15
3.16 Data Analysis ................................................................................................................ 16
™
3.17 Exiting the FX-5000
Program .................................................................................... 17
4. System Troubleshooting .................................................................................... 17
5. Application Notes .............................................................................................. 19
Sample Pressure Calculations Converting Sample Pressures to Forces ............................... 19
Baseplate and Gasket Cleaning and Disinfecting ................................................................. 19
6. Waveform Shape Definitions ............................................................................. 20
7. Flexcell
Appendix: FX-5000™ Compression System Dual Baseplate Operation
®
FX-5000™ Compression System Specifications
..................................... 22
.................. 24
i
FLEXCELL
®
INTERNATIONAL CORPORATION
ONE YEAR LIMITED PARTS WARRANTY
Flexcell® FX-5000™ Compression System
1.
FLEXCELL INTERNATIONAL CORPORATION warrants to the original purchaser/customer all hardware components of the Flexcell FX-5000
Compression System serial #___________________________for a period of one year from the date of shipment to the purchaser/customer to be free from
manufacturing defects in workmanship or materials with the following exceptions, terms and conditions:
2.
3.
If technical advisory support service is not available through your distributor or reseller, for service contact warranty headquarters by phone or fax.
*Customers should contact Dell Inc. for all warranty questions or concerns with regards to the Dell OptiPlex computer and monitor.
a.
ITEMS EXCLUDED FROM THE WARRANTY ARE: Dell OptiPlex desktop computer and monitor*, software, disks, manuals and external
peripherals such as printers, mouse or track ball units, imaging devices, vacuum pumps, air tanks, electric voltage converters, compressors, surge
suppressers and all other accessory equipment.
b.
c.
d.
e.
f.
g.
GOVERNING LAW. The performance of the duties and liabilities of the parties under the terms and conditions of this Limited Warranty shall be governed in
all respects by the laws of the State of North Carolina, the United States of America.
APPLICATION OF STATE LAWS: Some states do not allow the exclusion or limitation of consequential damages nor do some states allow limitations on
how long an implied warranty lasts, so the above limitations may not apply to you. This warranty gives you specific legal rights and you may also have other
rights that vary from state to state.
INTERNATIONAL CUSTOMERS. The full text of the foregoing limited warranty and all disclaimers is applicable to international customers/purchasers
except when the purchase was made from an international distributor or reseller, the warranty will be covered through your distributor or reseller.
DURING THE WARRANTY PERIOD, the purchaser/customer must notify Flexcell of any warranty claim in writing, by telephone, fax transmission
or email identifying each defective part or specifically describe the exact problem no later than the last day the warranty is in effect.
FLEXCELL AGREES to correct any defect in workmanship or material and supply new or rebuilt parts in exchange for defective parts upon
completion and submission by purchaser/customer of a printed “Parts Return Authorization” form furnished by Flexcell. Parts must be properly
packed in original container and shipped to our factory service center or distributor with all shipping costs prepaid if the unit is out of warranty
coverage. If the original shipping box is not available, Flexcell will send the required protective shipping container. (Flexcell will recommend the
insurance value for parts or equipment to be shipped.) Return carrier shipping costs within the US and Canada will be paid by Flexcell from the
service center. All other shipping costs outside of the US and Canada will be paid by the purchaser/customer. The purchaser/customer is solely
responsible for payment of custom fees, taxes, holding fees or value added taxes.
THIS LIMITED WARRANTY only covers failures due to defects in materials or workmanship which occur during normal use. It does not cover
damage which occurs in shipment or failures of original equipment due to products identified as add-ons not manufactured by Flexcell International
Corporation or its distributors nor does this limited warranty cover damages or failures which result from accident or disaster such as fire, explosion,
flood, wind, lightning, or earthquake or misuse, abuse, neglect, mishandling, misapplication, alteration, faulty installation, modification or service
by anyone other than our factory or distributor. Breaking the seal and opening the unit case or violating unit components will void the warranty. This
warranty is extended only to the original purchaser/customer unless a transfer of ownership is approved by Flexcell in writing.
LIMITED LIABILITY. Flexcell or its distributor’s only liability shall be to remedy any defect to comply with its warranty and return the repaired
equipment to function as designed. Under no circumstances shall Flexcell or its distributors be liable for any special incidental or consequential
damages based upon breach of warranty or contract or negligence. Such damages include, but are not limited to: loss of profits, revenue, loss of data,
down time, customer’s material or time.
DISCLAIMER OF WARRANTIES: The Limited Warranty expressed in the foregoing language is the only warranty applicable to this product. Any
other warranty, expressed or implied warranty or of merchantability or fitness for a particular purpose are hereby disclaimed. No oral or written
information or advice provided by Flexcell, through its agents or employees, in the use and functioning of the equipment shall in any way create a
warranty or in any way increase the scope of this limited warranty.
DISCLAIMER: LANGUAGE. This warranty document, accompanying instruction manual and supplemental applicable laws appear in the English
language. In the event of any inconsistency in the meaning of the words and terminology and any foreign language translation, the English language
shall prevail.
Within the United States only: 1-800-728-3714 (toll free) - Fax: 1-919-732-5196
International Customer’s telephone: 01-919-732-1591 - Fax: 01-919-732-5196
Email: info@flexcellint.com
FLEXCELL INTERNATIONAL CORPORATION
2730 Tucker Street, Suite 200, Burlington, NC 27215
1-800-728-3714 (within the U.S. only) 1-919-732-1591 fax: 1-919-732-5196 http://www.flexcellint.com
ii
FLEXCELL
®
INTERNATIONAL CORPORATION
Dell COMPUTER WARRANTY INFORMATION
Dear Customer:
Congratulations on your purchase of the Flexcell FX-5000 system. This system comes with a one (1) year
warranty. The Dell computer that is included with this equipment will be transferred into the Principal
Investigator and University’s name at the time of purchase. This computer comes with an excellent
warranty through Dell. If you have any problems with this computer, you will contact Dell directly. The
details of the warranty are listed below. You will need to have the service tag number and Express Service
Code number, which are located on the side of the computer.
Dell Hardware Limited Warranty Plus Onsite Service Extended Year
Dell Hardware Limited Warranty Plus Onsite Service Initial Year
Thank you choosing Dell ProSupport. For tech support, visit http://support.dell.com/ProSupport or
call 1-866-516-3115
ProSupport for IT: Next Business Day Parts and Labor Onsite Response 2 Year Extended
ProSupport for IT: Next Business Day Parts and Labor Onsite Response Initial Year
ProSupport for IT: 7x24 Technical Support for certified IT Staff, 2 Year Extended
ProSupport for IT: 7x24 Technical Support for certified IT Staff, Initial year
iii
FLEXCELL
®
INTERNATIONAL CORPORATION
FEDERAL COMMUNICATIONS COMMISSION
RADIO FREQUENCY INTERFERENCE STATEMENT
This computing equipment has been type-tested and found to comply with the limits for a Class B digital
device in accordance with the specifications for Subpart J of Part 15 of FCC Rules. These limits are
designed to provide reasonable protection against harmful radio and TV interference in a residential
installation. There is no guarantee that interference will not occur in a particular installation.
The following instructions and precautions must be observed by the user when installing and operating this
device:
1.
The equipment must be operated in strict accordance with the manufacturer's instructions.
2.
The unit must be plugged into a properly-grounded power outlet and used with the power cord
supplied with the unit, unmodified.
3.
The unit must be operated with the factory-installed cover in place.
4.
No modifications of the equipment can be made which may result in the equipment failing to meet
the limits of the FCC Rules.
5.
The equipment must be maintained in a satisfactory state of repair.
6.
Shielded cables must be used with this equipment to ensure compliance with the FCC Rules.
If the unit does interfere with radio or television reception, which can be tested by turning the unit off and
on, the user is encouraged to try the following suggestions to correct the interference:
1.
Reorient the antenna of the receiver.
2.
Relocate the equipment to increase the distance between the unit and the radio/TV.
3.
Connect the unit to an outlet on a different circuit from the receiver.
4.
Consult with the dealer or an experienced radio/TV technician for advice on ways to reduce
interference.
iv
FLEXCELL
®
INTERNATIONAL CORPORATION
FX-5000 Q
1. Choose a location for the FX-5000
UICK START INSTRUCTIONS
™
Compression system that is near the
incubator.
2. Place the computer, monitor, keyboard,
mouse, and FX5K™ Compression FlexLink
®
on a sturdy, flat surface.
3. Connect the keyboard, mouse, and monitor to
the computer according to the manufacturer’s
instructions.
4. Connect the FlexLink
®
(COMM) to the
Ethernet port on the back of the computer
using the Ethernet cable supplied (see Figs. 2
and 3, p. 2).
5. Connect the computer, monitor and
FlexLink
®
to a power source using a surge
protector.
6. Connect the system pressure line to the
®
SYSTEM port on the back of the FlexLink
using the ⅜" (9.5mm) OD blue tubing.
7. Cut the SYSTEM tubing near the unit and add
the pop-off valve and pressure gauge
assembly in line with the tubing.
8. Cut a 10' (3 m) length of the ¼” (6.4 mm)
blue and ⅜” (9.5 mm) clear tubing and
connect each to the back of the FlexLink
®
in
the FLEX IN and FLEX OUT ports,
accordingly. Attach the provided quick
disconnects to the other end of each piece of
tubing.
9. Place the tissue/gel samples onto the foam
disks in the center of each well of the four
BioPress
™
compression plates. Add 3-4 mL
of medium into each well. Place a stationary
platen into each well of the plates and screw
the center piece down until it touches the top
of the tissue/gel sample. Center the four
BioPress
™
plates on the four black
compression gaskets in the baseplate.
10. Adjust the clamping pads to their maximum
height on the compression clamping system.
Be sure that each pad surface is parallel with
the surface of the BioPress
™
baseplate to
prevent any of the pads from catching the
plate lids during insertion. Place the
Plexiglas
evenly along the bottom four clamping bars.
NOTE: This window is used as a backing
support for the BioPress™ baseplate to
enable it to withstand high pressures. Never
operate the compression baseplate without
this window in place.
11. Slide the BioPress™ baseplate/plate assembly
into the clamping system and adjust so that
the four plates are centered underneath the
eight clamping pads. Check the Plexiglas
window below and move as necessary to
center it under the BioPress
Tighten the pads down until the wing nuts are
finger tight. At this point the BioPress
plates should be sufficiently clamped.
NOTE: If using less than 4 culture plates in
an experiment, put an old plate or support in
place of the missing plate(s) and tighten the
clamping pads onto this surface.
12. Place the baseplate/clamping system
assembly into the incubator. Insert the ¼"
(6.4 mm) blue and ⅜" (9.5 mm) OD clear
tubing into the access port of the incubator.
Snap the quick disconnects for the ¼" (6.4
mm) and ⅜" (9.5 mm) tubing to the quick
disconnects on the baseplate.
13. Turn the FlexLink
ON. The Windows
on the monitor. Double click on the FX5000
to start the software.
14. In the Configure menu, select User Editor
and register as a user.
15. In the Regimen menu, select Create new and
create a regimen by entering values into the
appropriate spaces or click on the Run
Wizard button in the bottom left-hand corner
for a step-by-step procedure. When
completed, click on Save Regimen. Exit the
regimen editor.
window into the clamping system,
™
baseplate.
®
and computer system
desktop should appear
™
icon on the left hand side of the screen
™
v
FLEXCELL
®
INTERNATIONAL CORPORATION
16. In Controllers menu, select the appropriate
FlexLink
®
number to which the regimen will
be downloaded.
17. In the Regimen menu, select Assign. Choose
the proper platform, user, and regimen
(Important: see section 3.7 - Assigning
Regimens) from the drop-down menus.
Confirm with a Yes to download the regimen.
18. Once the program is downloaded, click on
Simulate in the main window display. Once
satisfied that the regimen is correct, click on
Stop.
19. Turn ON the pressure source. **Do not
allow the input pressure to exceed 20 psi or
the transducers may be damaged.
20. Click on Start to run the regimen. The plates
should begin flexing.
21. To pause or stop the regimen, click on Pause
or Stop at any time. Otherwise, the program
will stop once the regimen is complete.
vi
FLEXCELL
®
INTERNATIONAL CORPORATION
1. FLEXCELL® FX-5000 COMPRESSION SYSTEM COMPONENTS
1. Dell OptiPlex desktop computer.
Do not delete any software or change any
settings on this computer. It has been preconfigured to operate the FlexSoft® FX5000™ software.
Preinstalled software:
FlexSoft
®
FX-5000
™
Microsoft Windows® 7
Microsoft Office 2010
Adobe Acrobat
2. FX5K Compression FlexLink®
3. USB Keyboard
4. USB 2-button entry mouse
5. 17” Flat Panel monitor.
6. Accessory pack - nuts, ferrules
7. Flexcell® positive pressure baseplate
8. Gaskets and clamping system
9. Plexiglas® baseplate bottom support
10. Inlet and outlet tubing
11. Flexcell® FX-5000 Compression
System User Manual and other ancillary
manuals for computer and monitor
12. Surge protected outlet strip
13. Pop-off valve/Pressure gauge assembly
14. Positive pressure source (Not supplied)
2. SETUP OF THE FLEXCELL® FX-5000™ COMPRESSION SYSTEM
Do not power up the Flexcell® FX-5000™
Compression system until all connections
among components have been properly
made and checked! Never power up the
FX5K™ Compression FlexLink® with the
cover removed - an electrical shock hazard
exists. Operation of the computer without
the cover is in violation of FCC regulations
and will void the warranty!
2.1 P
ACKING LIST
Remove the packing list and check off the
instrument components as they are removed
from the shipping containers. If any parts are
missing, contact Flexcell
®
International Corp.
immediately.
2.2 L
OCATION OF THE
C
OMPRESSION SYSTEM
FX-5000
Choose a location for the FX-5000™
Compression system (Fig. 1) that is adjacent
and as close as possible to the CO2 incubator
and positive pressure source. A shorter
distance in the pressure lines decreases the
airflow resistance for compression and
release of the flexible bottomed plates,
allowing for a greater range of compression
at higher frequencies. The length of the
pressure line should be a minimum of 4’ (1.2
m). All cables (mouse, monitor, keyboard,
and Ethernet communications cables) will
connect to the rear of the computer (Fig. 3).
All tubing and the Ethernet communications
cable connect to the rear of the FX5K™
Compression FlexLink® (Fig. 2). The
length of the clear and blue tubing from the
®
FlexLink
to the baseplate should be 10' (3
m) long. This length can vary if necessary
™
Copyright 2009 Flexcell International
FLEXCELL, FlexLink and FlexSoft are trademarks of Flexcell International Corporation. MS Windows and MS Office are trademarks
of the Microsoft Corporation. Adobe Acrobat is a trademark of Adobe Systems Inc. Dell OptiPlex is a trademark of Dell.
General Notice: Other product names mentioned in this manual are for identification purposes only and may be trademarks of their respective
owners. Flexcell International disclaims any and all rights in those marks.
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FLEXCELL
®
INTERNATIONAL CORPORATION
but no more than 5' (1.5 m) longer or shorter.
Cut the tubing after the system is fully set up
to ensure of the proper length.
NOTE: The PUMP CONTROL CONNECTION
(Fig. 2) feature is only available with the FX5000™ Tension system.
mouse in front. The total weight of the
system is 48.37 lbs (21.96 kg).
Figure 1. Flexcell® FX-5000™ Compression
System
Figure 2. FX5K™ Compression FlexLink® rear
view
™
The FX-5000
Compression system will
require a space 28.75” (73.03 cm) wide by
22.44” (57.00 cm) deep if the computer,
™
FX5K
Compression FlexLink® and monitor
are placed side by side with the keyboard and
Figure 3. Dell OptiPlex rear view
2.3 S
ETUP OF THE COMPUTER AND
FX5K™ C
OMPRESSION FLEXLINK
®
Setup the Dell OptiPlex desktop computer
and peripherals according to the
manufacturer’s instructions. Refer to
Figures 1-10 for proper setup of the FX5000™ system instruments. Place the
FX5K
™
Compression FlexLink® next to the
computer and connect the FlexLink® to the
computer via the Ethernet cable provided.
®
If additional FlexLinks
have been
purchased, they are to be placed side by side.
For multiple FlexLinks
®
to be controlled by
one computer, connect the Ethernet cable
from each FlexLink® to an Ethernet Hub.
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FLEXCELL
®
INTERNATIONAL CORPORATION
Then, connect the hub to the Ethernet port on
the back of the computer.
Connect the system tubing as follows:
1. Connect the ⅜” (9.5 mm) O.D. blue tubing
from the pressure source to the SYSTEM
port on the back of the FlexLink® (Fig. 1
& 2).
2. Attach the free end of the clear ⅜” (9.5
mm) tubing to the FLEX OUT port on the
back of the FlexLink® (Fig. 2).
3. Attach the free end of the blue ¼” (6.4
mm) tubing to the FLEX IN port on the
back of the FlexLink® (Fig. 2). The VENT
port should be left open.
NOTE: When connecting the tubing to the back
of the FlexLink®, slip the brass nut with the nylon
ferrule over the tubing, then push the tubing onto
the fitting as far as possible and tread the nut onto
the fitting. Tighten each nut by hand until the
tubing is completely crimped. If the tubing is not
firmly connected it may blow off under high
pressures.
Connect the power cables (115 or 230 VAC)
to the rear power inlet (FlexLink Power) of
the FlexLink® (Fig. 2). If the available power
supply is 115 VAC or 230 VAC, be sure the
voltage selection switch ON the computer
(Fig. 3) is set to 115 V or 230 V, respectively.
When the instruments are completely set up,
the power cords should be connected to a
surge protected power strip and then to a 115
VAC or 230 VAC, 60 Hz power outlet.
2.4 I
NCUBATOR ACCESS PORTS
Consult the operations manual of the CO2
incubator for the location of the incubator
access ports. Most incubators have an access
port at the top or the rear of the incubator. If
this opening is not immediately apparent,
consult the incubator manufacturer about the
location of an access port. To prevent voiding
the incubator warranty or encountering an
electrical hazard, a trained incubator
representative should open the incubator
access ports.
Once the access port has been located, insert
the FLEX IN and FLEX OUT tubing
through the access port into the incubator.
Attach the ¼” (6.4 mm) blue tubing to the
¼” (6.4 mm) quick disconnect on the
baseplate. Attach the ⅜” (9.5 mm) clear
tubing to the ⅜” (9.5 mm) quick disconnect
on the baseplate (Fig. 4). If two baseplates
are to be used simultaneously, connect the
baseplates using the configuration as shown
in Figure 5.
Figure 4. Single baseplate connection
(clamping system not shown)
Figure 5. "T" connection for two baseplates
(clamping systems not shown)
3
FLEXCELL
2.5 S
AMPLE PREPARATION AND SETUP
BIOP
OF
AND
A
SSEMBLY
RESS™ CULTURE PLATES
B
ASEPLATE CLAMPING
®
INTERNATIONAL CORPORATION
The BioPress
™
compression culture plate
contains a foam ring and sample holder in
each well (Fig. 6). The foam ring is used to
contain the removable sample holder. In
preparing samples, be sure that the sample
thickness is no less than 1000 m and no
greater than 0.118” (3 mm). The thickness of
the compressed foam is 350 m; therefore,
thinner samples will not be compressed. A
sample thickness of 1000 m or more will
ensure compressibility. The maximum
chamber height is 0.122” (3.1 mm); therefore
samples with a thickness greater than 0.118”
(3 mm) will be preloaded when the platens are
inserted into the wells of the culture plates.
To prepare a sample for loading, remove the
sample holder from the plate using a pair of
sterile forceps. Place the tissue or gel sample
into the hole at the center of the removable
sample holder. Carefully place the sample
holder back into the BioPress
™
compression
plate so that it is contained within the foam
ring. Do this for the remainder of the wells.
Add 3-4 mL of culture medium to each well.
Prepare all stationary platens by adjusting
the center screw (Fig. 7) such that the screw
bottom (part that touches the sample) is
exactly flush with the bottom of the platen
body. Use a pair of sterile forceps and the
two holes at the adjustable center of the
platen to turn the center screw.
Figure 6. Compression culture plate foam ring
and sample holder
The sample diameter should be equal to or less
than the inner diameter of the foam sample
holder in the chamber, which is 0.200” (5
mm). If the user wishes to use a larger sample,
they can remove the sample holder and place
the sample within the larger ring that normally
contains the sample holder. This will allow a
sample diameter of up to 0.525” (13 mm).
Figure 7. Stationary platen and compression
culture plate
Place a stationary platen into each well of the
compression plate (Fig. 7). Calculate the
number of revolutions required to bring the
bottom of the center screw to the point where
it just touches the top of the sample by using
the following equations:
For sample height in millimeters,
x = (3.11-h) / 0.62
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FLEXCELL
®
INTERNATIONAL CORPORATION
where x is the # of 180 degree clockwise turns
required for the center screw to touch the
sample and h is the sample height in
millimeters.
For sample height in inches:
x = (0.122-h) / 0.025
where x is the # of 180 degree clockwise turns
required for the center screw to touch the
sample and h is sample height in inches.
NOTE: If the sample height is larger than 3.11
mm (0.122”), ‘x’ in the above equations will be a
negative number. In this case, the bottom of the
center screw will compress the sample when it is
flush with the bottom of the platen body as the
platen is first inserted into the well. To
compensate for this, the center screw can be
turned counterclockwise ‘x’ number of turns;
however, it should be noted that the sample will no
longer be fully compressible, as the base of the
sample holder will no longer be able to contact the
base of the center screw.
Using sterile forceps, turn the center screw
clockwise the number of turns required by the
above equation(s). Be careful not to screw the
center down too far as this will preload the
sample and give inaccurate force readings.
Add or remove fluid medium through any of
the eight holes around the inner periphery of
the platen (Fig. 7).
™
Center the BioPress
compression plates on
top of the four black gaskets in the baseplate
(Fig. 8). Adjust the eight clamping pads in the
clamping system to their maximum height and
turn them so that the bottom of each pad is
parallel with the surface of the baseplate.
Place the Plexiglas window into the
clamping system (Fig. 9), evenly along the
bottom four clamping bars
NOTE: The Plexiglas window is used as a
backing support for the baseplate to enable it to
withstand high pressures. Never operate the
compression baseplate without this window in
place.
Baseplate without plates
Baseplate with plates
Figure 8. Baseplate with compression plates
Figure 9. Clamping system with Plexiglas®
window
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FLEXCELL
®
INTERNATIONAL CORPORATION
Carefully slide the baseplate/plate assembly
into the clamping system so that the four
BioPress™ plates are centered underneath the
eight clamping pads (Fig. 10). Check the
Plexiglas
window and move as necessary to
center it under the baseplate. Turn the wing
nuts on the clamping pads until they are finger
tight. This will compress the plates downward
onto the gaskets, creating a seal.
Baseplate slides into
clamping system (Plexiglas
window on bottom) and then
tighten the clamps down over
the compression plates. Do
not over tighten.
Plexiglas window must fit
under the baseplate to
keep seal.
Figure 10. Clamping system with baseplate
and culture plates
NOTE: If using less than 4 plates in an
experiment, use an old plate or support in place of
the missing plate(s) and tighten the clamping pads
onto this surface. This will ensure that the
baseplate is seeing an even and sufficient pressure
across its surface and will prevent baseplate
blowout.
Place the entire assembly into the incubator.
Attach the FLEX IN and FLEX OUT quick
disconnect fittings to the fittings on the
compression baseplate (Fig. 1 & 4).
2.6 C
ONNECTING THE SYSTEM
P
OSITIVE PRESSURE LINES
Connect a minimal length of the blue ⅜” (9.5
mm) tubing to the positive pressure source.
The pressure source should be capable of
maintaining a pressure of 12-15 psi (83-104
kPa) when the FlexLink® is running. Most
house pressure systems will be capable of
maintaining this pressure. If a floor pump is
to be used, the following specifications are
recommended as a minimum:
Max Pressure: 60 psi (414 kPa)
Max Free Airflow Rate: 1.1 cfm (31
L/min)
If the pump does not meet one of the above
specifications, there is still a possibility that
it will have the capability of providing the
necessary pressure and flow rate
combination to run the FX-5000™
Compression System. If the pump is capable
of producing a minimum of 0.9 cfm (25.5
L/min) at 12 psi (83 kPa), the pump can be
used with the FX-5000™ Compression
System. Check with the pump manufacturer
to determine if the pump is capable of
meeting these specifications.
NOTE: Flexcell® highly recommends the use of
a pressure reservoir when using a floor pump as
the positive pressure source. Contact Flexcell®
for information on purchasing a pressure
reservoir.
Keep the length of tubing between the
pressure source and FlexLink
®
as short as
possible (4’ or 1.2 m minimum). If the
pressure source does not have an adjustable
regulator built into it, a pressure regulator
will likely required to keep the input
pressure to the system at an acceptable level
(recommended at 12-15 psi (83-104 kPa),
no greater than 20 psi [140 kPa]). Most
house pressure systems will produce
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pressures up to several hundred pounds per
square inch (1 MPa or more). Such pressure
levels will damage the pressure transducers
in the FX5K™ Compression FlexLink®.
To be sure that the proper input pressure level
is achieved, when running the system, first
adjust the pressure level to 15 psi (103 kPa)
when the unit is not yet running a regimen.
This can be done using a pressure regulator, or
by adjusting the relief valve on the pressure
output side of the pump (if using a small floor
pump). Once the regimen has begun, check to
see how much the pressure drops on the
gauge. Readjust the pressure regulator or
relief valve as necessary to get the input
pressure between 12 and 15 psi (83 and 104
kPa) while the regimen is running. This will
assure a proper pressure level for the unit to
function correctly.
NOTE: With optimal pressure settings, there will
still be some system limitations due to airflow.
Combinations of high frequencies and forces may
not be achievable due to the high pressure level in
the short period of time requested. An example
would be a regimen programmed to run from 0-12
pounds at 1.0 Hz. The system is not able to achieve
the pressure required to reach 12 pounds in 0.5
seconds because of airflow limitations. Therefore
the output may only reach 9 or 10 pounds. The
exact output is dependent on several system
variables such as tubing and overall system
volume. To maximize system performance, use the
shortest tubing lengths possible (see section 2.2).
2.7 F
AND
LEXSOFT®
S
YSTEM POWER-UP
FX-5000
SOFTWARE
The FlexSoft® FX-5000™ software
(Flexcell Copyright Protected Software,
Property of Flexcell® International
Corporation, 2730 Tucker Street, Suite
200, Burlington, NC 27215 has been preinstalled on the hard drive.
Keep the CD package included with the
system and store it in a safe place. Be sure
to shutdown Windows properly by using
the Shut Down command activated by the
Start button. Failure to follow the above
directions can cause software and
hardware malfunctions.
The computer has Windows XP installed
on the hard drive. All backup software is
provided with the system. Check all
connections and connect the surge protector
into an appropriate power outlet (115 VAC
or 230 VAC). Turn ON the surge protector
strip and check that the indicator light on the
strip is lit. If the power light does not come
on, the surge protector has failed. Replace
the surge protector and attempt to determine
what caused the surge protector to fail. Turn
ON the power switch on the computer,
monitor and FX5K™ Compression
FlexLink®. The system should then boot and
load Windows®. Select and double click on
the FX-5000™ icon.
NOTE: Flexcell® recommends that the power
saving or screen saver features in the Windows
software are disabled. Allowing the hard drive
to power down while an experiment is running
may create the need to reboot the computer
before data can be viewed, or before a regimen
can be downloaded again. It is recommended
that the monitor be turned OFF when not in use
and that the computer be allowed to run at full
power during an experiment.
2.8 LCD D
C
OMPRESSION FLEXLINK
ISPLAY ON THE
FX5K
™
®
The front of the FX5K™ Compression
FlexLink® box has an LCD graphic display.
™
When the FX5K
Compression FlexLink®
is turned ON, the LCD will display the
FlexLink
®
number (i.e., #1, #2, #3, or #4)
and the regimen status (i.e., Idle, Ready, or
Left (the time remaining, in minutes, of a
running regimen). When a regimen has been
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downloaded and/or is running, the LCD will
also display the user name, platform, and
regimen name. See Table 1, below, for the
display messages with various FlexLink®
statuses.
Table 1. Messages displayed on the FX5K™
Compression FlexLink
FlexLink Status LCD message
Power ON #1 Regimen: IDLE
Regimen
downloaded
Regimen is
running
or being
simulated
Regimen has
stopped
®
LCD.
Flexcell International Corp.
www.flexcellint.co
#1 Regimen: READY
User name
Platform
Regimen Name
Flexcell International Corp.
www.flexcellint.co
#1 Regimen: LEFT 5 min
User name
Platform
Regimen Name
Flexcell International Corp.
www.flexcellint.co
#1 Regimen: IDLE
User name
Platform
Regimen Name
Flexcell International Corp.
www.flexcellint.co
2.9 U
SING ADDITIONAL FLEXLINKS
®
Using additional FX5K™ Compression
FlexLinks® will allow two or more baseplates
(platforms) with different regimens to run
simultaneously. All tubing connections made
to the first FlexLink® should be made exactly
the same with any additional FlexLinks®.
Each FlexLink® will have its own separate
FLEX IN and FLEX OUT tubing for each
baseplate. However, the pressure or
SYSTEM tubing can be shared between
FlexLinks
®
with "T" fittings as long as the
pressure source is capable of running two or
more FlexLinks® with the programmed
regimens. Regimens with a combination of
high frequencies and force levels will
require a more powerful pressure source.
The capability of the pressure source with
more than one FlexLink® will need to be
tested experimentally with the particular
setup. FlexLinks® must be placed side-byside during use.
®
To connect additional FlexLink
(s) to the
computer, connect the Ethernet cable from
each FlexLink
®
to an Ethernet hub. Then,
connect the hub to the Ethernet port on the
back of the computer using the Ethernet
communications cable provided. The LCD
on the front of the FlexLink® will display the
number (#1-4) that the FX5K™ Compression
FlexLink® has been assigned at the time of
production.
NOTE: The numbering of each FX5K™
Compression FlexLink® will not depend on what
order it appears in the connection chain, but
according to the configuration of each
FlexLink® as "FlexLink #1, FlexLink #2…etc"
during production.
When powering up the FlexLink®(s) and
computer, turn the FlexLink®(s) ON before
the computer. To use a particular FlexLink
select the appropriate FlexLink® number
(#1, 2, 3, or 4) from the Controllers menu.
Assign, download, and run regimens just as
would be done when using one FlexLink
®
,
®
.
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3. OPERATION INSTRUCTIONS
3.1 S
TARTING THE
FX-5000
™
PROGRAM
The Flexcell system should now be set up
and ready for operation. Be sure that the
system positive pressure supply is adequate to
operate the instrument(s) (see section 2.6).
Turn ON the power switch on the computer,
monitor and FX5K™ Compression FlexLink®.
The system should then boot and load
Windows®. Select and double click on the FX5000™ icon (Fig. 11) on the desktop to start
the FlexSoft® FX-5000™ software. A main
display window will appear (see Fig. 13).
Figure 11. FX-5000™ icon
3.2 U
SING THE
FX-5000
™
TUTORIAL
In the Help menu, click on View Tutorial to
view the FX-5000™ tutorial at any time (Fig.
12). The tutorial will appear on the left side of
the screen. The tutorial explains the main
steps involved in configuring users, creating
regimens, and starting regimens. The tutorial
will start automatically, as a default setting,
when the software is opened. To turn OFF this
feature, uncheck the box marked “Show
Tutorial at Startup”.
3.3 M
AIN DISPLAY WINDOW
The main display window (Fig. 13) for each
™
FX5K
Compression FlexLink® has several
features that allow more precise monitoring of
the regimen. To access this window, select the
appropriate FlexLink
®
(FlexLink#n) from the
Controllers menu, where “n” is the
®
FlexLink
be downloaded. The FlexLink
number to which the regimen will
®
number can
be found on the LCD screen on the front of
the FlexLink® controller box.
Figure 12. FX-5000™ tutorial
Figure 13. Main display window
All of the remaining information and
command button states in the main
window display reflect the current state of
the system for the currently selected
FX5K
™
Compression FlexLink®.
It is possible to simultaneously assign and
run a different regimen for each of the
®
attached FlexLinks
.
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In the upper left hand corner of the window is
a section labeled Plot Units. This function
allows users to change the units of the y-axis:
force (lbs) or kPa. To select a particular unit,
click on the appropriate button beside the unit
label.
The next block to the right is a section labeled
Smoothing. One expects data patterns to
represent normal distributions of a function
describing a straight line, sinusoidal curve,
etc. Therefore, the plotted points of a
mathematical function should be a predictable
data set that is “smooth” (i.e., a curve
comprised of points whose coordinates are
predicted by the function). For a straight line,
the equation would be y = mx + b; for a power
function, the equation may be y = x2 or some
derivative thereof. In general, data patterns
may be confounded with noise. For the case of
the pressure plot as a function of voltage from
the FX-5000™ Compression System pressure
transducer, the rough data yield a plot with a
coefficient of variation of only 0.01%.
Therefore, the signal to noise ratio for the
combined Flexcell® system of circuits,
hardware and transducers is high, lending
confidence in the displayed plot.
Nevertheless, the user can select a data
smoothing option that is a simple weighted
average of the points. The Smoothing section
contains the option of either applying no
smoothing (None) to the data points returned
®
from the FlexLink
or of applying a threepoint moving average filter (Moving
Average) to the incoming data. Choose the
desired affect by clicking on the appropriate
button.
Below the box for Smoothing is a section used
to show the current X and Y coordinates for
the waveform. These are displayed when the
left mouse button is clicked and held down in
the graph area. When this is done, a pair of
crosshairs appears and the coordinates at the
intersection of the crosshairs are shown in the
X and Y spaces. The crosshairs and values
will track the mouse as long as the left mouse
button is held down.
The next block to the right is a section used
to display the current User, Regimen,
Platform, and Run time. Run time has two
blocks. The one on the left displays the
elapsed regimen time, and the one on the
right displays the remaining regimen time,
using the following format: dd:hh:mm:ss
(days:hours:minutes:seconds).
The block at the top right contains the
current regimen Step, Cycle, Jump (when a
regimen step is using the Jump Back to
option), minimum level (Min), and
maximum level (Max). The minimum and
maximum levels will be listed according to
the units specified under the Plot Units
section as the minimum and maximum
points at which the membrane stretches with
each cycle.
The major portion of the main display
window is taken up with a plot designed to
show the current state of a simulated or
running regimen. This plot is generated from
the data returned by the FlexLink
®
during
regimen activity. When using a platform
configuration that uses positive pressure to
generate an upward movement of the
membrane, resulting in compression, the
vertical scale represents increasing
compression/force. When the selected unit
of display is kPa, the vertical scale will start
at 0.0 and move upwards as an increasing
positive level, indicating that positive
pressure is being applied to the membrane to
compress it upwards against an immovable
barrier, the platen.
The scale on the x- and y-axes can be
changed by clicking on one of the two small
double-arrow buttons at either end of the
axis. The button with arrows pointing away
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from each other increases the axis scale, while
the button with arrows pointing toward each
other decreases the scale.
Below the plot is a timeline that displays the
percentage of the regimen that has been
completed.
To the right of the plot is a set of command
buttons which allow the user to:
1) Start a regimen.
2) Stop a regimen.
3) Pause a regimen.
4) Resume a paused regimen.
5) Simulate a regimen.
6) Reset the controller.
7) Access Help.
8) Cancel the program.
3.4 C
ONFIGURING USERS
Prior to creating and downloading a regimen,
a user needs to be registered. Once this task is
completed, regimens can be personalized
under the registered user name.
In the Configure menu, select User Editor. A
new window will appear entitled Users (Fig.
14).
To add a user, type in a user name (must be at
least 4 characters) in the provided space and
click on Add User. The new name should
appear in the Existing Users box.
To delete a user, click on the appropriate user
name in the Existing Users box so that it is
highlighted in blue, then click on Delete User.
The program will ask for a confirmation and
issue a warning, click Yes to confirm deletion.
The user and any of the user's defined
regimens will be deleted.
To exit the Users window, click on Close.
Figure 14. FX-5000™ user editor
3.5 C
REATING REGIMENS
Before beginning, a regimen must be created
for downloading. In the Regimen menu,
select Create new. A new window will
appear entitled Regimens with a list of the
existing users and their regimens (Fig. 15).
In the Existing Users box, click on the user
name under which the regimen is to be
created. A list of current regimens created
under this user will appear in the Regimens
for Selected User box, unless no regimens
have been saved under this user, in which
case, this box will be empty.
Figure 15. FX-5000™ regimen editor
To create a new regimen, click on Run
Wizard in the bottom left hand corner. This
option will bring up a step by step process of
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creating a new regimen. Alternatively, follow
the sequence of steps below:
1) Type a regimen name in the box labeled
Regimen Name.
2) Choose the desired waveform shape under
Shape by clicking on the downward arrow
and selecting an entry from the drop-down
list.
3) Enter the desired minimum and maximum
force (in lbs) for the plate membranes (015 lbf) in the boxes under Min and Max.
4) Enter the frequency for the waveform
(0.01-5 Hz) in the box under Freq.
5) If a static regimen is required, enter the
desired force at which the membrane is to
be held in the Max box. Enter 0 in the Min
box, and 1.0 in the Freq box (Min and
Freq are irrelevant for static regimens).
6) If a duty cycle for the waveform is to be
defined, enter the desired value in the box
under DC %. The duty cycle is the percent
of time that the waveform shape remains
on at the high (square wave) or rising
(triangle) part of the waveform. Duty
cycle only affects the triangle and square
waves.
7) Enter the number of times for the regimen
to repeat (0-65,535) in the box under
Cycles. When moving out this box, the
box underneath dd:hh:mm:ss will update
to the calculated regimen time according
to the set parameters. The format for this
entry is days:hours:minutes:seconds.
However, if the time is entered first, the
program will calculate the number of
cycles.
8) To make a regimen step (except for the
first step) return (jump back) to a previous
regimen step, enter the return step
number in the box under Back To. Then,
type the number of times to repeat this
process in the box under Repeat.
For instance, if the user was entering
Step 4 and wanted to return to Step 2 and
repeat Steps 2 through 4 three times,
enter 2 under Back To and 3 under
Repeat when inputting Step 4
9) When the regimen step is complete, click
on Add Step at the bottom. Every time a
new/edited step is saved, the overall
duration of the regimen will be
calculated and displayed in the box
labeled Duration next to the Step
number box. If the total duration exceeds
the maximum allowable duration, this
value will be MAXIMUM, and the
regimen cannot be saved. Adjust the
time and/or cycle number to decrease the
total duration time.
10) To add other steps to a regimen, click in
the box labeled Step and type in the next
step number.
For example, the first step will
automatically be Step 1. To add another
step, delete the 1 and replace it with a 2.
After this is completed, change the
parameters as needed for the new step
according to steps 2 to 9 above.
11) To delete a step that has already been
added, click on the step in the window
labeled Regimen Steps, which contains
the list of steps. This will cause the step
to be highlighted. Then click on Remove
Step at the bottom of the window.
Confirm the deletion by clicking on Yes
when prompted.
12) After all steps and parameters have been
entered, click on Save Regimen.
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13) To delete a regimen, simply double-click
on the desired regimen name in the
Regimens for Selected User box, and
then click on Delete Regimen. Confirm
the deletion by clicking on Yes.
NOTE: If the regimen to be deleted is currently
assigned, the regimen must be removed from
assignment before deleting.
14) When finished with regimen editing, click
on Close.
3.6 C
REATING A REGIMEN WITH A
C
USTOM WAVEFORM SHAPE
In the top row of an Excel spreadsheet, enter
the force values for the desired waveform
shape for one cycle. Enter one value per cell.
Values should be chosen along a curve at even
time intervals, i.e. every 100 msec
(milliseconds). Save this spreadsheet as a .csv
file (comma delimited file format) in the
C:/FlexSyst directory.
Example Custom waveform shape: Ramp
waveform shape
6
5
4
3
% Elongation
2
1
0
0 100 200 300 400 500 600 700 800 900 1000
Time (msec)
Example Excel file: Ramp waveform shape
1 1.5 2 2.5 3 3.5 4 4.5 55.50
Example CSV file viewed in Notepad: Ramp
waveform shape
1,1.5,2,2.5,3,3.5,4,4.5,5,5.5,0
In the Regimen menu of the FX-5000™
software, select Create new. A new window
will appear entitled Regimens with a list of
the existing users and their regimens. In the
Existing Users box, click on the user name
under which the regimen is to be created. A
list of current regimens created under this
user will appear in the Regimens for
Selected User box, unless no regimens have
been saved under this user in which case, this
box will be empty.
To create a new regimen, click on Run
Wizard in the bottom left hand corner. This
option will bring up a wizard to create a new
regimen. Or, follow the sequence of steps
below:
1) Choose Custom from the list of
waveform shapes under Shape by
clicking on the downward arrow and
selecting this entry from the drop-down
list.
2) Choose the appropriate file name under
File by clicking on the downward arrow
and selecting an entry from the dropdown list
3) Type a regimen name in the box labeled
Regimen Name.
4) Enter the time interval in milliseconds
between two force values in the box
under Time.
5) Enter the number of times for the
regimen to repeat (0-65,535) in the box
under Cycles.
6) When the regimen step is complete, click
on Add Step at the bottom. Every time a
new/edited step is saved, the overall
duration of the regimen will be
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calculated and displayed in the box
labeled Duration next to the Step number
box. If the total duration exceeds the
maximum allowable duration, this value
will be MAXIMUM, and the regimen
cannot be saved. Adjust the time and/or
cycle number to decrease the total
duration time.
7) Click on Save Regimen.
8) When finished with regimen editing, click
on Close.
3.7 A
SSIGNING REGIMENS
In the Controllers menu select the
appropriate FlexLink#n, where “n” is the
FlexLink® number to which the regimen will
be downloaded.
In the Regimen menu, select Assign, then
choose the Platform, User, and Regimen
from each drop down menu (Fig. 16). A
message box will appear to confirm regimen
assignment. Click Yes.
NOTE: Be sure to choose the platform that
properly corresponds to the setup being used.
For the
System, the BioPress and StagePresser platforms
will be used depending on the purchased device.
An error message will be given if an incompatible
platform is selected.
Flexcell® FX-5000™ Compression
If the download progresses, no further
messages will appear, and the Start and
Simulate buttons will be enabled. If an error
occurs during the downloading of the regimen
to the FlexLink®, a message will alert the user
of a problem, and they will not be able to run
the regimen. In which case, click on Reset in
®
the FlexLink
window, and then assign the
desired regimen again.
Figure 16. Assigning regimens for
downloading to the FX5K
FlexLink
™
Compression
®
.
3.8 S
TARTING REGIMENS
To run an assigned regimen, click on Start
in the main display window (Fig. 13) after
the regimen has finished downloading. Be
sure that the positive pressure source is ON
and all other connections are made. The
membranes should begin flexing and the
assigned regimen will be in progress.
The progress of the regimen for the currently
selected FlexLink® will be monitored
continuously. The displays for Run Time,
Step #, Cycle #, Jump #, and Min and Max
% will update, as will the actual waveform
®
data returned by the FlexLink
.
Additionally, a progress bar below the graph
will display the percentage of the regimen
that has been completed.
3.9 S
IMULATING REGIMENS
The Simulate button is used to simulate a
regimen graphically without actually
running it. Simulating a regimen will give
the user an idea of the waveform output
before actually running the regimen. To start
a simulation, click on Simulate in the main
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display window. A waveform will begin
moving across the graphic display. This
waveform corresponds to the ideal waveform
as generated by the FX-5000™ software, and
is the actual waveform pattern sent to the
FlexLink® during the download process. For
certain waveforms (e.g. square waves), the
idealized pattern may appear to have some
unusual elements (e.g. rounded corners). The
purpose of these unusual elements is to
accommodate for the lag time during sudden
changes in pressure (e.g. rising/falling edge of
a square wave). Without these unusual
elements, the valve controllers may oscillate
for a period of time (e.g. dampened sine wave
during beginning of plateau in a square
waveform).
3.10 P
AUSING REGIMENS
To pause a simulated or running regimen,
click on Pause in the main display window.
Click on Yes when the program asks for
confirmation. Pausing will cause the current
simulation or actual running of the regimen to
stop at the next cycle of the current regimen
step. Pausing is different from stopping, in
that pausing leaves the regimen at its current
position, whereas stopping causes the regimen
position to be set to the beginning of the first
regimen step.
3.11 R
ESUMING REGIMENS
To resume a paused regimen, click on
Resume in the main display window. This
function is available only when a simulated or
running regimen has been paused. The
regimen will restart its prior activity
(simulation or actual run) from its last point.
3.12 S
TOPPING REGIMENS
To stop a simulated or running regimen, click
on Stop in the main display window. Click on
Yes when the program asks for confirmation.
Stopping causes the current activity to cease
immediately, and sets the regimen position
back to the beginning of the first regimen
step. Stopping is different from pausing, in
that pausing leaves the regimen at its current
position so that it can be resumed from
where it stopped.
3.13 D
ETERMINE THE PROGRESS OF
R
THE
EGIMEN
The progress of the regimen for the currently
selected FlexLink® will be monitored
continuously. The displays for Run Time,
Step number, Cycle number, Jump to
number, and Min and Max force will be
updated in the upper right hand corner of the
main display window. The actual waveform
data returned by the controller will also be
continuously updated on the graph. A
timeline progress bar below the plot
illustrates the percent of the regimen that has
been completed.
3.14 R
ESETTING THE FLEXLINK
®
Resetting a simulated or running regimen
causes the current activity to stop at the next
cycle of the current regimen step, and
disassociate the regimen from the FlexLink®
controller. To reset, click on Reset in the
main display window. Click on Yes when
the program asks for confirmation. The
system will now be in state similar to that
which exists when the FX5K™ Compression
FlexLink® is first turned ON.
3.15 U
SING THE PRESSURE/FORCE
C
ONVERSION CALCULATOR
To convert pressure (kPa) to force (lbs) or
vice versa, select Calculate from the
Regimens menu. To select the Convert
Pressure to Force or Convert Force to
Pressure calculator (Fig. 17), click on the
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appropriate button beside the calculator label.
Figure 17. Calculator for converting pressure
to force, or vice versa.
For pressure to force conversions, enter the
pressure in kPA in the Pressure (kPa) box
and the sample diameter in millimeters in the
Sample Diameter (mm) box. Click
Calculate. The calculated force in lbs will
appear in the Force (lbs) box.
For force to pressure conversions, enter the
force in lbs in the Force (lbs) box and the
sample diameter in millimeters in the
Sample Diameter (mm) box. Click
Calculate. The calculated pressure in kPA
will appear in the Pressure (kPa) box.
To exit this window, click on Close.
3.16 D
ATA ANALYSIS
The FX-5000™ software has the capability of
storing data (a set of waveform points) from a
regimen run to a file on the hard drive. The
filename consists of the FlexLink
®
number, a
“V” or “C” code letter (V for vacuum, or C for
compression), the regimen name and a
sequence number. If a compression regimen is
ran three times on FlexLink#1, then there will
be three data files on the hard drive with
numbers ‘000’, ‘001’ and ‘002’ (i.e.,
1_C_regimenname_001.dat,
1_C_regimenname_002.dat, etc.). The file
size depends on the regimen duration. The
current settings take pressure data every 2
milliseconds (i.e., 500 points/sec or
1,800,000 points/hour). Therefore, a one
hour regimen will have approximately a 4.0
Mb data file. To turn this feature OFF, please
contact Flexcell®.
To see the stored data, in the Data Analysis
menu, select Data Viewer. Open the
appropriate .dat file from the C:/FlexSyst
directory. Two new windows will appear
(Fig. 18). In the window labeled filename:1,
the data presented defaults to the raw data.
In the window labeled filename:2, the data
presented defaults to a moving average of
the raw data with data averaged every 10
seconds.
Figure 18. FX-5000™ data display
The y-Axis control box allows for automatic
or manual control of the scale on the y-axis.
If Auto is selected, the y-axis scale can be
changed by clicking on one of the two small
double-arrow buttons at either end of the
axis. The button with arrows pointing away
from each other increases the axis scale,
while the button with arrows pointing
toward each other decreases the scale. The
minimum value is -2 kPa and the maximum
is -100 kPa. If Manual is selected, the user
can enter the upper and lower limits of the yaxis scale in the white boxes to the left side
of the axis. The small double-arrows will not
function if Manual is selected.
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The x-axis scale allows for a time interval to
be set between 0.1 and 1000 seconds. Click on
one of the two small double-arrow buttons at
either end of the axis to change this scale. The
button with arrows pointing away from each
other increases the axis scale, while the button
with arrows pointing toward each other
decreases the scale.
The Input data box allows the user to switch
the data in the graph from raw data to
averaged data. If the Raw data option is
selected, the graph will plot the pressure in
–
kPa over time, in seconds, for the selected
regimen. If the Moving average option is
selected, the graph will plot the average
pressure over the designated time period (in
seconds, range of 0.1-1000 seconds) shown in
the Time (s) box.
The Start time box can be set so that the
data viewed in the plot begin at the specified
time entered here. Use the timeline at the top
of the graph to navigate through the regimen.
Users can move the slider function within
the timeline to manually progress through
the regimen plot. If the user clicks on the
arrows at either end of the timeline, the plot
will progress to the next time period as set
by the x-axis scale. For instance, if the scale
is set to show 20 seconds of data and the user
clicks on the right arrow button, the plot will
progress to the next 20 seconds of data.
3.17 E
P
XITING THE
ROGRAM
FX-5000
™
To exit the FX-5000™ software, in the
Controllers menu, select Exit.
4. SYSTEM TROUBLESHOOTING
NOTE: If your FX-5000™ system is not
functioning properly and the solution is not
clearly outlined or defined below, please contact
Flexcell® before proceeding with any repair work.
Problem 1: Gaskets on baseplate are not
sealing
Symptoms:
a. System is not performing to specified
parameters; i.e., not reaching
maximum %elongation entered in
regimen
b. Baseplate is making a whistling or
blowing sound when air pressure is
added
Test to determine if this is the problem:
If you are experiencing either of the above
symptoms, there is likely a leak somewhere in
the system. First, check all tubing to be sure
that it is straight and not bent to restrict air
flow. To find the location of pressure leaks,
do the following:
Solution 1:
Check the Flex In and Flex Out
connections on the back of the FlexLink®
unit. Be sure that these are connected
properly and sealed tightly. Next, run a
regimen and take note of the max and min
forces shown on the screen. Disconnect the
quick disconnects from the baseplate and
then connect them together, bypassing the
baseplate. Take note again of the max and
min forces. They should increase slightly
because of the decrease in system volume. If
the force outputs on the screen are now
reaching the forces that you requested in
your regimen, then the leak is likely in your
baseplate/clamping system. If the force
output remains incorrect with respect to your
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FLEXCELL
®
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regimen inputs, check your input pressure
level from your pressure source. Try
increasing it slightly. If the problem remains
then the FlexLink® may need recalibration.
Call Flexcell® International for further
instructions regarding recalibration.
To correct a leak problem in the baseplate, try
increasing the clamping pressure on the
BioPress compression plates. If this doesn't
work, remove the clamping pressure from the
BioPress plates. Remove the BioPress
plates from the gaskets and check the
indentation made by the plates on the gaskets
to be sure that the plates were approximately
centered on the gaskets (i.e., not sitting on an
area near the end of the gasket width). Reseat
plates as necessary and reclamp as before.
NOTE: In many cases it may not be possible to
get a perfect seal between the culture plates and
gaskets. When you are not able to reach your
maximum requested force because of a baseplate
leak problem, try increasing your input pressure
from your pressure source. Be careful not to
exceed 20psi.
Problem 2: Regimen wave is flatlining
Solution 2:
Remove the SYSTEM pressure line at the
back of the controller and check to be sure that
there is pressure coming through the line.
Check the VENT port at the back of the
controller when the system is running to see if
air is moving in or out. If not, it is possible that
the valves are locked due to particle
accumulation over time. Try applying air
pressure to the vent port to free the valves. If
the regimen wave is flatlining but the plates
are still cycling air pressure, try the following:
Disconnect the blue ¼" (6.4mm) FLEX IN
tubing from the back of the unit and place your
finger over the end of the tubing while a
regimen is running. If air is cycling at the end
of the tubing then the problem is with the
FlexLink®. Contact Flexcell® International
for further instructions.
Problem 3: Regimen wave is fluttering or
not smooth
Solution 3:
This problem is due to a miscalibration
of the FlexLink®. Contact Flexcell® for
further instructions.
Problem 4: You want to program a long
regimen (1 day or more) and the software
will not allow you.
Solution 4:
The software has a limit on the number
of cycles that can be entered into any given
step (65,535; see section 3.2.5, #7 of the
manual). If you program a regimen with a
step that requires more than 65,535 cycles,
the software will give you an "invalid entry"
error. To run the regimen for the time
required you will need to make an additional
step. This will not create any breaks in the
regimen. Simply divide your total time in
half (or as many times as is necessary) and
make additional steps with the same
parameters. See section 3.2.5 of the manual
for instructions on programming additional
steps.
Problem 5: “An invalid argument was
encountered" error is shown when
attempting to assign a regimen.
Solution 5:
This problem is caused when a user
name is entered into the software without
any regimens listed under his/her name. If
the assign function sees that a user has
nothing to assign an error is displayed.
Simply delete the user(s) without a regimen
or make and save a regimen under the user(s)
name(s).
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FLEXCELL
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Problem 6: One or more of the FX5K™
Tension FlexLinks® connected to the
system are not being recognized by the
software. This is usually represented by the
error message “Cannot Connect to
Controller” when attempting to connect a
FlexLink® to assign a regimen.
Solution 6:
This problem is usually due to a lack of
communication between the computer and
one or more FX5K™ Tension FlexLinks®.
5. APPLICATION NOTES
Check the FX5K™ Tension FlexLink®
Ethernet port connection on the computer
and the communications connection on the
back of the FlexLink® to be sure that these
are secure. Check the FlexLink® to see if the
power is ON (the FlexLink screen should
power on). If both of these conditions are
true, close and restart the software. Try
downloading the regimen again. If the
problem is not resolved, call Flexcell® for
further instructions.
S
AMPLE PRESSURE CALCULATIONS
C
ONVERTING SAMPLE PRESSURES TO
F
ORCES
The actual pressure applied to a sample in the
well of the BioPress™ compression plate is
dependent on the surface area of the sample to
which the force is being applied. Therefore,
the FX-5000™ Compression System is
designed to apply a force to 3-D samples so
that force is not dependent on sample size.
To convert a pressure to a force, input the
desired pressure to be applied to the sample
into the equation below, along with the sample
diameter, and program the resulting force into
®
the FlexSoft
software.
Force
= 0.177*(P
lbs
MPa
)*(D
mm
2
)
where Force
sample or cell in pounds, P
is the force applied to a single
lbs
is the pressure
MPa
to be applied to the sample in megapascals
(MPa), and D
sample in mm.
is the diameter of a single
mm
Given the force programmed into the
software, the pressure applied to the samples
given can be calculated using the following
equation:
P
= (5.65* Force
MPa
lbs
)/(D
mm
2
)
where P
sample in megapascals (MPa), Force
is the pressure applied to the
MPa
lbs
is the
force entered into the regimen or displayed
on the software screen in pounds, and Dmm is
the diameter of a single sample in mm
B
ASEPLATE AND GASKET
C
LEANING AND DISINFECTING
Neither the black urethane nor the black
ABS (acrylonitrile butadiene styrene)
baseplate should be autoclaved as the high
temperature could cause warping. The
baseplate is not dishwasher safe because the
high water temperature may cause warping.
The black rubber gasket should not be
subjected to the high temperatures involved
with autoclaving and or in dishwashers.
Both baseplate and gasket should be cleaned
with regular laboratory detergent, rinsed
with warm tap water, followed by distilled
or deionized water and allowed to dry. The
baseplate should be dried and stored on a flat
surface. Do not hang the black gasket on
pegs to dry as the gasket can be stretched.
Protect both pieces from UV light exposure.
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FLEXCELL
®
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To disinfect, use a mild bleach (commercial
bleach solution such as "Clorox" or other
laundry bleach solution is normally 5.25%
sodium hypochlorite) solution (1:500, bleach
to water) or a 1% benzalkonium chloride
solution, which can be sprayed, poured or
wiped on both pieces and allowed to stand on
surfaces for at least 5 min. Rinse with distilled
or deionized water. The above solutions are
effective against yeast and fungi. A 70%
ethanol solution also can be applied and
pieces allowed to air dry, do not wipe dry.
Tubing on the baseplate may be flushed with
70% ethanol, but be sure tubing is completely
dry before connecting to the Flexcell Unit.
Contact time (time surface remains wet) of the
above solutions is critical to the effectiveness
of disinfecting. Times given are minimal and
longer contact times are recommended in
cases where contamination is severe.
The above decontamination procedure
should be performed before and after each
use of the baseplate and gasket or whenever
media is spilled onto these surfaces. Be sure
that the openings and the channels in the
baseplate are dry and clear of any
obstructions before using.
Do not store baseplate or gasket in
incubator as the warm, humid conditions
will promote growth of microorganisms.
After cleaning and disinfecting, seal
baseplate and gasket in a plastic bag. Store
baseplate flat and be sure that the entire
bottom of baseplate is supported to prevent
warping. Store gasket away from UV light
exposure.
6. WAVEFORM SHAPE DEFINITIONS
½ Sine Wave
This waveform provides a smooth, physiological
variation in pressure at the max% with a quick
transition at the min%.
Sine Wave
This waveform provides a smooth, physiological
variation in pressure from min% to max%.
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FLEXCELL
®
Triangle Wave
This waveform provides a linear-ramped rise to the
max% and a linear-ramped fall back to the min%.
INTERNATIONAL CORPORATION
Square Wave
This waveform provides the fastest possible rise to
max%, followed by the fastest possible fall to min%.
Heart (E) Wave
This waveform applies a pressure pattern that mimics
the shape as seen on an electrocardiogram
Static Wave
This waveform applies a constant state of pressure.
Heart (P) Wave
This waveform applies a pressure pattern that mimics
the actual pressure wave generated by a heartbeat.
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FLEXCELL
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7. FLEXCELL® FX-5000™ COMPRESSION SYSTEM
SPECIFICATIONS
FX-5000
FX-5000
FX-5000
FX5K
™
COMPRESSION SYSTEM FEATURES
Uses air pressure to deform a flexible-bottom culture plate yielding up to 15 pounds of applied force to a
tissue or 3D cell culture sample
Minimum force resolution capability is 0.1 lb force
Applies a defined, controlled, static or cyclic deformation to growing cells in vitro
Housed valving mechanism automatically regulates and maintains pressure to provide the specified
compression regimen
Capable of driving up to 4 independent FlexLink
remote compression and/or strain controllers
Capable of delivering frequency ranges from 0.015 Hz
™
Works with BioPress
series culture plates
Able to program multiple frequency, amplitude and wave changes in one regimen
Waveforms available: Static wave, Sinusoidal wave, Heart wave (Electronic and Pressure), Triangular wave,
Square wave and custom waveforms
™
Optional: StagePresser
device allows compression application to cells while viewing in real time under an
upright microscope
™
COMPRESSION COMPONENTS AND SPECIFICATIONS
Dell OptiPlex Desktop Computer
USB Keyboard
USB Mouse
17" Flat panel Monitor
®
FlexSoft
FX5K
FX-5000™ V1.0 (Flexcell FX-5000 Software), Microsoft Windows 7
™
Compression FlexLink controller
Accessory Pack - nuts, ferrules
Flexcell
positive pressure baseplate, gaskets, clamping system and Plexiglas baseplate bottom support
Inlet and outlet tubing
Flexcell
FX-5000 Compression System User's manual, Culture Plate & Loading Stations™ manual and
other ancillary manuals for computer and monitor
Surge protected power outlet strip
Pop-off valve/Pressure gauge assembly
™
COMPRESSION COMPUTER SPECIFICATIONS
Dell OptiPlex Small Form Factor
Intel Core i5 processor (3.2 GHz)
250 GB SATA 6.0 Gb/s hard drive
DVD +/- RW Drive
2 GB of DDR3 RAM
®
Integrated video, Intel
HD2500
Operating System:
Windows 7 Professional
™
COMPRESSION FLEXLINK COMPONENT SPECIFICATIONS
Custom design positive pressure controller board
Microprocessor sampling rate for display data transducer: 200 Hz
Microprocessor sampling rate for feedback/error (valve adjustment) transducer: 1 KHz
2 proportional-solenoid valves
Ethernet port (RJ45)
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FLEXCELL
®
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FX5K
™
COMPRESSION FLEXLINK® CALIBRATION SPECIFICATIONS
Static Performance:
Maximum allowable waveform variance from static line: +/- 0.15 lbs
Maximum force error (actual vs. programmed): +/-0.35 lbs
Maximum allowable transducer inaccuracy: +/-2.0 kPa
Dynamic Performance:
Maximum force error (actual vs. programmed): +/-0.35 lbs, at the following
Frequencies: < 10 lbs, 0.5 and 1.0 Hz; 10 < x < 14 lbs, 0.5 Hz
Maximum allowable transducer inaccuracy: +/-2.0 kPa
FX-5000
FX-5000
Power Requirements: 115/230V –8.8/4.3A or 4.3/2.2A – 60/50 Hz
Weight (unit only): 15.0 lbs (6.8 kg)
FX5K
Power Requirements: 115/230V – 6/3A – 60/50 Hz
Weight (unit only): 22.15 lbs (10.06 kg)
Monitor 17”LCD Flat Panel: Size, W x H x D: 14.6” x 15.65” x 7.1” (37.1 x 39.75 x 18.0 cm)
Power Requirements: 115V - 60 Hz Auto switch to 230 V
Weight (unit only): 11.22 lbs (5.1kg)
™
COMPRESSION PHYSICAL SPECIFICATIONS
™
Computer: Size, W x H x D: 3.65” x 12.45” x 13.40” (9.3 x 31.6 x 34.0 cm)
™
Compression FlexLink®: Size, W x H x D: 10.5” x 7.0” x 18.5” (26.7 x 17.8 x 46.9 cm)
FX-5000
To achieve the maximum system capability, the recommended compressed air requirements are:
™
COMPRESSION - COMPRESSED AIR REQUIREMENTS
The pressure source must be able to produce 12 PSI (83 kPa) of pressure at 0.9 cfm (25.5L/min).
If a house pressure system is being used and there are no means to test the flow rate capabilities, run the
house pressure system with the FX-5000
consistently while the FlexLink
™
the FX-5000
is running then your house pressure source is sufficient.
®
is running a regimen. If it is able to maintain 12-15 PSI (83-104kPa) while
™
to determine if it is able to maintain 12-15 PSI (83-104kPa)
The following is an example of specifications for a floor compressor pump that can supply enough pressure
™
to run the FX-5000
. The pressure versus flow curve for this pump shows that it can achieve the necessary
12 PSI (83 kPa) of pressure at 0.9 cfm (25.5L/min). Contact the manufacturer for the specifications of the
floor compressor pump to determine if it will meet the necessary requirements.
o Maximum Pressure: 60 PSI (414 kPa)
o Maximum Free Airflow Rate: 1.1 cfm (31 L/min)
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FLEXCELL
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APPENDIX: FX-5000™ COMPRESSION SYSTEM
DUAL BASEPLATE OPERATION
When using the FX-5000™ Compression System in dual baseplate operation, the user may notice
a drop in the overall programmed Force, depending upon the desired Force and frequency of the
regimen. This drop is due to the pressure source being shared between both baseplates.
It is, therefore, recommended that users reduce the frequency of such a regimen to 0.5 Hz or lower.
A lower frequency will allow the FX-5000™ Compression System the correct amount of time to
intake and exhaust the air, which, in turn, gives the system enough time to reach the desired Force.
For example, if an FX-5000
configuration and a regimen is programmed with the following variables:
Waveform = Sine
Force = 15 lbs
Frequency = 1 Hz
The maximum Force achieved might only be 10 lbs. Reducing the frequency of the regimen to
0.5 Hz will produce an achievable Force of around 13 lbs.
™
Compression System is connected in the dual baseplate
NOTICE
The information in this document is subject to change without notice. Flexcell® International
assumes no responsibility for any errors that may appear in this guide. This manual is believed to
be complete and accurate at the time of publication. In no event shall Flexcell
®
International
Corporation be liable for incidental or consequential damages in connection with or arising from
the use of the manual. Opening the case of the FX5K
warranty. Customers must contact Flexcell
®
International Corporation or an authorized
distributor before opening the case on the FX5K
™
Compression FlexLink® voids the
™
Compression FlexLink®.
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