BioTester

Biaxial Test System

User Manual
version 6.5
Mechanical measurement and analysis of biomedical materials

CellScale provides scientific and medical researchers with turn-key systems for
measuring the mechanical properties of biomaterials.
Our focus is soft tissue – including skin, ligaments, blood vessels, heart valves,
sclera, membranes and scaffolds. We provide user-friendly software, an easy-to-
use patented attachment system, and effective data analysis tools.
Our foundation was laid at one of the world’s leading research institutions – The
University of Waterloo. We understand research and aim to provide effective
solutions at a reasonable price.
Explore our web site www.cellscale.com or contact us to learn more about our
measurement systems.

© 2014 CellScale. All rights reserved. This material may not be reproduced,
displayed, modified or distributed without the express prior written permission of the
copyright holder. For permission, contact CellScale Biomaterials Testing at
[email protected].

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Canada N2L 5C6
Phone: 519.342.6870
Table of Contents
1. General Information .................................................................................................................. 1
Environmental and Electrical Specifications ................................................................................ 1
System Assembly......................................................................................................................... 1
Power Connections ...................................................................................................................... 1
Safety Warnings ........................................................................................................................... 1
Approvals and Certification .......................................................................................................... 1
Manual Operating Controls .......................................................................................................... 2
General Maintenance ................................................................................................................... 2
2. Testing Terminology ................................................................................................................. 3
Multiphase Test Cycles ................................................................................................................ 3
Phases, Cycles, and Test Sequences ......................................................................................... 4
Test Phases: The Smallest Unit of Testing .................................................................................. 5
Control Modes .............................................................................................................................. 5
Control Functions ......................................................................................................................... 6
3. System Overview ..................................................................................................................... 7
System Components .................................................................................................................... 7
Software Overview ....................................................................................................................... 8
Output Files and Data Structures................................................................................................. 9
4. Setting Up & Starting a Test ................................................................................................... 10
Overview .................................................................................................................................... 10
Step 1: Start a New Test ............................................................................................................ 10
Step 2: Reset the Actuators (occasionally) ................................................................................ 11
Step 3: Move the Actuators to a Specified Size ......................................................................... 11
Step 4: Zero the Load Cells (occasionally) ................................................................................ 12
Step 5: Modify Testing Parameters (optional) ........................................................................... 12
Step 6: Mount the Specimen ...................................................................................................... 14
Step 7: Execute the Test ............................................................................................................ 15
Step 8: Terminate the Test Prematurely (optional) .................................................................... 15
5. Additional Settings .................................................................................................................. 16
Configuring Output Data Files.................................................................................................... 16
Advanced System Settings Dialogue ......................................................................................... 17
Configuring the Live Charting Graphs ....................................................................................... 19
6. Reviewing Test Results .......................................................................................................... 21
Overview .................................................................................................................................... 21
Selecting Images........................................................................................................................ 22
 Image Playback Options ............................................................................................................ 22
Image Tracking: Overview ......................................................................................................... 23
Image Tracking: The Points Display Option .............................................................................. 25
Image Tracking: The Displacement Option ............................................................................... 26
Image Tracking: The Strains Option .......................................................................................... 27
Data Overlay .............................................................................................................................. 28
Exporting Tracked Data ............................................................................................................. 29
Exporting Images and Movies.................................................................................................... 30
7. System Hardware Settings ..................................................................................................... 31
Controller .................................................................................................................................... 31
Camera ...................................................................................................................................... 31
Load Cells .................................................................................................................................. 32
Actuators and Motors ................................................................................................................. 32
Temperature............................................................................................................................... 33
External Sync Pulse ................................................................................................................... 34
8. System Calibration and Advanced Tools ............................................................................... 35
Snap Image Feature .................................................................................................................. 35
Load Cell Calibration .................................................................................................................. 35
Adjusting the Camera Position and Image Magnification .......................................................... 37
Update Firmware........................................................................................................................ 39
9. Troubleshooting ...................................................................................................................... 40
10. Appendix A: Initial System Setup ....................................................................................... 41
Tools .......................................................................................................................................... 41
Fasteners & Parts ...................................................................................................................... 41
Unpack System .......................................................................................................................... 42
Attach Camera Mast .................................................................................................................. 43
Attach Magnets to Goosenecks ................................................................................................. 44
Place Fluid Chamber on Riser Stage ........................................................................................ 45
Install Camera Lens ................................................................................................................... 45
11. Appendix B: Software Installation ....................................................................................... 46
12. Appendix C: Install Load Cells / Perform Camera and Gooseneck Alignment .................. 52
Connect USB cables to the PC USB 2.0 ports and then proceed with the following steps. ...... 52
Attach the Load Cells to the Goosenecks .................................................................................. 52
Attach the Goosenecks to the Actuators ................................................................................... 54
Plug in the Load Cells ................................................................................................................ 56
Launch the LabJoy Software ..................................................................................................... 57
Perform Camera Alignment ....................................................................................................... 60
 Perform Vertical Gooseneck Alignment ..................................................................................... 61
Perform Transverse and Axial Gooseneck Alignment ............................................................... 63
Install Actuator Covers ............................................................................................................... 66
13. Appendix D: Protocol for Tying and Cutting Sutures for Biaxial Testing ............................ 67
 BioTester User Manual

1. General Information
The BioTester is a precision test instrument specifically designed for the measurement and
analysis of small biological specimens. The system includes a compact biaxial test station and an
integrated software interface to run and analyze test results. The system is intended for indoor
use only.

Environmental and Electrical Specifications

Electrical Input 100-240VAC, 50-60Hz
Current Rating 2.5 Amp
Environmental Conditions Maximum Operating Temperature 25ºC
0% - 95% Relative Humidity
Installation Category Category II
Pollution Degree Degree 2
Maximum Altitude 2000m
Data Connections 1 – USB 2.0 for camera/PC communication
1 – USB 2.0 for controller/PC communication

System Assembly
The unit requires some assembly. See Appendix A and C.

Power Connections
Connect power supply into properly grounded 100-240VAC power source to ensure safe
operation. Ensure that the power cord is easily accessible at all times. The use of an
Uninterruptible Power Supply (UPS) is recommended to protect against data loss.
The mains supply voltage fluctuations should not exceed 10% of the nominal supply voltage.

Safety Warnings
This equipment must be used in accordance with the procedures outlined in this manual to
prevent injury and/or damage.

System Alert
The plastic covers must remain on this unit at all times to prevent injury and/or damage.
This equipment must not be disassembled by the user or modified in any way.

Approvals and Certification

This product conforms to EN61010-1:2001 and EN61326-1.

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Manual Operating Controls

There are a few manual controls on the unit. The camera housing contains two adjustment knobs
that are used for fine adjustment of the camera, and the camera lens has a focus ring and an iris
which can be set by the user.

BioTester Tip: Setting Camera Controls

The camera housing knobs are used to ensure that the specimen is centered in the field
of view. The iris is usually left 100% open; the brightness of the images is better
adjusted by changing the camera gain and exposure.

There are two switches on the front of the unit, one for powering the control board and the LED
lighting, and the second for powering the heating element.
The front of the unit also contains a pull rod which is used to raise and lower the specimen stage
and fluid chamber. This rod is spring-loaded so that the chamber can be held at any height. To
raise or lower the stage, lift the rod and then pull or push.

General Maintenance
Clean the system as needed with mild soap and water or alcohol based cleaning solutions.

BioTester Tip: Removing the fluid chamber

The fluid chamber can be removed by resetting (fully retracting) the actuators followed
by carefully lifting and twisting the chamber. This is best accomplished when the
chamber is emptied (a large syringe works well).

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2. Testing Terminology
The BioTester is designed to apply biaxial forces to soft tissue specimens (deforms by more than
1% in vivo) with an in-plane dimension of 3 - 15 mm. This includes biological material such as eye
tissue, heart valves, pericardium, joint capsules, large blood vessels, scaffolds, and polymers.

Multiphase Test Cycles

In order to properly characterize and test a specimen, it is often necessary to load it to different
degrees and at different rates. There are three main reasons for doing this:
Preconditioning - The goal of preconditioning is to restore a specimen to its physiological or
in vivo state. During the process of specimen storage and preparation, a specimen may
swell, dry out, have its material fibers realign, or its molecules reorganize. It may take multiple
preconditioning cycles for a specimen to be restored to its natural state.
Reproducing Physiological Conditions During Testing - By applying various loads and
load rates, natural expansion and contraction of a specimen can be reproduced (for example,
the pulse pressure in an organ). In as much as the physiological conditions can be recreated,
the specimen can be tested in a more realistic state.
Varying Test Conditions - Variable loads and rates allow you to create a variety of test
profiles to best study your specimen.

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Phases, Cycles, and Test Sequences

As the following diagram demonstrates, each application and release of load on the specimen is
called a test cycle. The same test cycle can be repeated multiple times to achieve a certain goal
(preconditioning, physiological conditioning, or testing); this is called a test set. Finally, a test
sequence is made up of multiple test sets.

The above example describes the following:

 The entire diagram presents a full test sequence.
 Within that sequence, there are two test sets: the first set applies preconditioning to the
specimen; the second set executes the actual test on the specimen.
 Within the first set (preconditioning), two identical test cycles are implemented to bring
the sample to a satisfactory in vivo state.
 Finally, the second set (testing) is made up of three cycles.

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Test Phases: The Smallest Unit of Testing

The test phase is the smallest unit of the test specification. There are five phases within a cycle.
Each phase serves a specific purpose:
Preloading- Preloads are applied to bring a test
to a well-defined starting point. Because the
dimensions of a specimen may change as a
result of a loading cycle (stretching of fibers,
viscoelastic effects, plastic deformation, or
localized material failure at the attachment
points), the preload adjustment compensates for
any of these changes in specimen geometry.
Stretching - During the stretch phase, a
deformation is applied to the specimen. The
deformation can be specified either in terms of
force applied or displacement achieved.
Holding - The deformation can be held for a given duration. The duration for which it is
held is dependent on the nature of the testing.
Recovering - The recovery phase is the time during which the force being applied to
the specimen is removed. The duration of the recovery time is configurable and
dependent on the nature of the testing.
Resting - Finally, the rest phase is the time between the end of one cycle and the
beginning of the next. Some tests may specify a short recover time, while others may
specify a longer time. The duration is configurable and dependent on the nature of the
testing.

Control Modes
There are two control modes which define the basic approach to a given test: displacement
control and force control:
Under displacement control, the displacement of the specimen is predefined. The
BioTester stretches the specimen until the predefined displacement is achieved. The
force required to achieve the displacement is an output of the test.
Under force control, the force applied to the specimen is predefined. The BioTester
stretches the specimen until the predefined force is achieved. The displacement required
to achieve the force is an output of the test.

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Control Functions
The BioTester makes it possible to test specimens under several control functions:
Under displacement control:
The true strain function applies the displacement at a constant true strain rate, which
accounts for the current specimen length while the specimen is being stretched.
The ramp function applies the displacement at a constant nominal rate. This is
equivalent to a constant engineering strain or constant velocity.
The sine function applies the displacement according to a sinusoid with the desired
displacement magnitude and duration.
The custom load functions allow users to customize the rate and extent of
displacement application by specifying a table of time versus displacement values.

Displacement

Time

True Strain Ramp Sinusoid Custom

Under force control:

The step function achieves and maintains the desired force as quickly as possible. The
amount of time it takes to achieve the desired force depends on the material being tested
and the force control settings.

Force

Time
Step

The control mode, load function and load magnitude can be set independently for each of the two
loading directions.

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3. System Overview
System Components
The BioTester brings together several high-precision components, making it the state-of-the-art
instrument for testing biomaterials.

A High resolution CCD camera provides

synchronized video tracking and analysis.
B Illumination and lens provide excellent image
quality.
C Our patented BioRake attachment method
provides quick and easy mounting of samples.
D Integrated machine controls and data collection
are compact and designed for testing flexibility.
E High resolution control delivers precision
measurement on small samples.
F A simple USB interface for easy connection to
the host computer.

The BioTester

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Software Overview
The software included with the BioTester is called LabJoy. It is divided into two modules, a data
collection module and a review and analysis module. The data collection module is used to set
test parameters, enable specimen loading and testing, and monitor test progress. The screen
layout for this module is shown below:

Test
Parameters

Live Force &

Displacement

Live Video Live Charting

The review and analysis module is used to playback accelerated or decelerated test images,
perform image analysis and tracking, and output video files for presentation purposes. The
screen layout for this module is shown below:

Image List

Image Playback

Image Tracking

Data Overlay

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Output Files and Data Structures

For each test, the BioTester creates and saves three file types. The following table describes the
three file types for a project named “Sample1”. Output from this test would be found in a
“Sample1” output directory (a sub directory of the user specified data directory).

File Type Description

.tst file The Sample1.tst file that contains the exact protocol and settings used for the test.
.csv file The Sample1Data.csv file that contains comma separated numerical data such as time, force and
displacement values.
.jpg files Captured images such as Sample1.000010.3.jpg, which would correspond to an image captured
at 10.3 seconds from the start of the test.

Each test folder will also contain 2 subfolders. The “Logs” subfolder contains a text file of the
content of the text dialogue portion of the main screen. It is useful as reference to troubleshoot
problems with your system, should they occur.
The “Tracking” subfolder is initially empty. If tracking is performed on any of the images in the
test folder using the “Analyze and Review” software module, there will be data files that contain
the tracking information stored in this directory.
While using the “Analyze and Review” software module, you may create additional data files such
as text files (*.csv) containing tracking results, images with force data or tracking results overlaid
(*.wmf), or movie files (*.avi). These additional files can be stored in the test directory or
elsewhere on your computer’s hard drive without interfering with the software application (once
created, they cannot be opened by the LabJoy application).

When working with the data, you should be aware of a few details regarding the data output:
1. The output specimen sizes are based on the spacing between the BioRake tines and do
not account for the specimen material outside of the test region.
2. The software cannot calculate stresses since the thickness of the specimen is not known.
To calculate stress, you will have to manually measure the thickness of the material
before or after the test is performed.
3. Strains can be calculated using the output displacement values (which are based on the
BioRake tine motions). The specimen may actually be subjected to less strain than the
calculated values due to attachment site effects and tissue tearing. The image tracking
module is useful for determining the actual strain values and variations within the
specimen.

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4. Setting Up & Starting a Test

Overview
Setting up and running a new test is a simple process made up of both mandatory and optional
steps. The following list presents all of the steps, while the rest of this chapter describes each of
the steps in detail.
Step 1: Start a New Test
Step 2: Reset the Actuators (occasional)
Step 3: Move the Actuators to a Specified Size
Step 4: Zero the Load Cells (occasional)
Step 5: Modify the Test Parameters (optional)
Step 6: Mount the Specimen
Step 7: Execute the Test
Step 8: Terminate the Test Prematurely (optional)

Step 1: Start a New Test

Launch the LabJoy software, and then

select Collect New from the File menu.
In the Create Test from Template
dialogue, perform the following steps:

1. Select a template that matches the type of test you wish to perform. See the BioTester
Tip on how to select and use a template. You can modify the template parameters in
Step 4, below.
2. Name your test. The dialog will have a default test name. You can use the default or
rename the test. Each time you start a new test, the default name will continue to
increment the number at the end of the name.
3. If desired, you can change the location of the output data. The location of your output
data and images is determined by specifying a Test Name and Data Directory. The
template and data directories are user specific (computer login name). Each system user
can store their files to a different default location. The system uses the last location
selected as the default.

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4.

BioTester Tip: Selecting and Using Templates

How to select a template: Designing an appropriate test sequence is an art that
depends on both the type of material being tested and the specific material properties
you are interested in measuring. When first testing a new material, you should expect to
have to experiment with the settings until the test yields meaningful data.
The system comes loaded with example templates to help you get started. Selecting a
template does not lock you into a specific test sequence or protocol – rather a template
defines a test sequence and settings, all of which can be changed before a test is run.
Once you have developed a test sequence and settings that are appropriate for the
material you are testing, you should save these settings using “Save As Template” from
the file menu. You can then select your template the next time you initiate a test.

Step 2: Reset the Actuators (occasionally)

While it is not necessary to reset the actuators with every test sequence, we suggest performing
a reset at the start of a new test session. If you have stopped the previous test in mid cycle, then
you should reset the actuators. By resetting the actuators, you are moving the actuators to the
precise home position ensuring that the physical measurements taken by the BioTester are
accurate.

To reset the actuators, select Reset Actuator from the tools menu, or press on the toolbar.

System Alert
You should not reset the actuators if the specimen is already loaded. Doing so will
damage your specimen as well as possibly damage the BioRakes or Load Cells.

Step 3: Move the Actuators to a Specified Size

After the actuators have been reset, they will remain in their fully retracted positions and must be
moved to specified (reference or starting) positions.

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To move the actuators to the specified position, select Move Actuators to Specified Size from the
Tools menu, or press on the toolbar.

Step 4: Zero the Load Cells (occasionally)

While it is not necessary to zero the load cells with every test sequence, we suggest zeroing the
load cells at the start of a new test session. With repeated use, the zero point of the load cells can
drift. By zeroing the load cells, you are ensuring that the force measurements are accurate.

To zero the load cells, select Zero Load Cells from the tools menu or press on the toolbar.

System Alert
You should not zero the load cells if the specimen is already loaded, or the load cell
zeroing process will not work properly and an offset introduced into your force data.

Step 5: Modify Testing Parameters (optional)

You can select and modify parameter
sets by clicking on their row in the Test
Parameter Specification table and then
pressing the Edit Set button (or by
double clicking on their row). When you
do so, the Set Parameter Editor Dialog
will appear.
Note that displacements are specified
in either % strain, or μm. Force loads
are specified in mN, and durations are
specified in seconds.
The following table describes each of
the parameters.

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Test Parameter Description

Control Mode Biaxial tests are typically performed under displacement control, however, you can
select the following control modes to achieve specific testing objectives:
 To obtain uniaxial properties for a biaxially mounted specimen, specify
displacement control in one axis and Force control with zero Load Magnitude
in the other axis.
 For creep testing, use force control with a long hold duration.
 For relaxation testing, use displacement control with a long hold duration.
Control Function True Strain or Ramp is typically selected for tests performed in displacement control
mode. For tests performed in force control mode, you select step controls.
Stretch Magnitude Selecting a stretch magnitude is entirely dependent on the material you are testing. A
(Load Magnitude when in sound approach is to begin with a small magnitude and iteratively move up to larger
Force Control mode) magnitudes.
If you are using Displacement control mode, you can specify the displacement in either
µm or as a percentage. For example, the displacement on a 5000µm x
5000µmspecimen can be expressed as either 500µm or 10%.
If you are using Force control mode, you can only specify the force in mN.
Watch the test results carefully to determine which magnitude setting best achieves
your test goals.
Preload Preload is typically reapplied on every repetition during a preconditioning set as well as
on the first repetition of a testing set. Specimen size is adjusted after a preload
adjustment. Strain calculations are based on the specimen size after the last preload
adjustment. If you are working with a material for which preload values have been
suggested, you can set the value accordingly. Otherwise, zero is a good initial choice.
Preload Magnitude As with stretch magnitude, the preload magnitude settings depend on the material you
are testing. While Preload can be set at zero, typically you would set the preload
magnitude somewhere between zero and 10% of the peak load you expect to achieve.
Stretch Duration For evenly spaced images, it is recommended to choose a number that is an integer
multiple of the Image Output Frequency.
Hold Duration Hold Duration is typically set to 0, however it is useful for creep or relaxation testing.
Recovery Duration Recovery Duration is typically set to the same value as the Stretch Duration.
Rest Duration Rest duration is typically set to 0, however a non-zero value may be used to mimic in
vivo conditions or for specialized testing.
Repetitions Apply enough repetitions until the force deformation curves from one repetition to the
next start to overlie each other.
Data Output Frequency Typically set to the same frequency as the image output frequency.
Image Output Frequency Typically set to 1Hz for cycles > 5 seconds and 10 Hz for cycles < 5 seconds.

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Step 6: Mount the Specimen

Proper specimen mounting is critical to ensuring accurate and repeatable results. It is best to
have the tines spaced evenly across the edge of the specimen and as closely to the edges as
possible.
To mount the specimen, follow these steps:
1. From the tools menu, select Move Actuators to Specified Size, or press on the
toolbar.
2. Place the specimen in the mounting press on top of a backing material such as soft
silicon or foam.
3. Place the mounting press across the fluid chamber.
4. Partially raise the mounting press.
5. Centre the specimen between the rakes using a small
implement, such as forceps, and the live video window
as feedback. Adjustment of the actuators may be
necessary to ensure the rake tines are positioned
correctly for non-ideally sized specimens.
6. Raise the stage until the rake tines are pressing gently
on the specimen.
7. Push all 20 tines into the specimen, either all at once
using the press block or one at a time using a small
implement such as a precision screwdriver.
8. Partially lower the stage, and remove the backing
material.
9. Fully lower the stage, and remove the mounting bridge.
10. Fully raise the stage to submerge the specimen, if
desired. Typically, you will want to match a specimen’s
physiological state, which for most biological materials
is in a saline solution heated to the organism’s body
temperature.
11. To minimize the effect of surface debris and surface
ripples, a cut-out has been made on the top of the fluid
chamber that is sized to hold a standard microscope
slide.

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BioTester Tip: Ideal Specimen Size

The BioTester is designed for small specimen sizes. For our Standard BioRakes (5
tines spaced at 1 mm) the ideal specimen size is square with side lengths of 5.5 - 6 mm.
If a specimen is not in the ideal range, it may still be mounted, but the placement of the
rake tips will need to be adjusted.
Manual adjustment of rake positions during specimen loading may be necessary to
ensure that the rakes puncture the tissue correctly.
These manual adjustments result in a change in specimen size, which in turn results in
a change in actuator displacement profiles. The specified specimen size is displayed at
the top of the screen while the current specimen size is displayed on the left of the
screen just below the force output.
At any point before the test has started, specimen size can be changed using one of the
following methods:

 press on the tool bar, or from the Tools menu select Move Actuators to
Specified Size
 use the Actuator Control jog button arrows.

Step 7: Execute the Test

Select Execute from the File menu, or press the button on the toolbar.

Step 8: Terminate the Test Prematurely (optional)

You can stop a test at any time by clicking on the toolbar, or by selecting Stop from the File
menu. This is preferred over powering down the equipment, as it allows the software to store the
current actuator position and eliminates the need to reset the actuators on subsequent tests.
At the end of the completed test (both normally or prematurely terminated) the actuators will
maintain their current position.

BioTester Tip: Specimen Thickness

Measuring the specimen thickness before and/or after a test is performed will allow
stress calculations to be made from your output data.

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5. Additional Settings
Configuring Output Data Files
From the Settings menu, select Data Output to display the dialogue shown on the left. Click the
Configure button to select which of the following columns to output.

Output Column Description

Set Name User defined name of each set
Cycle Cycle and Phase information
Time (S) Time in seconds
X/Y Size (µm) Distance between opposite pairs of BioRakes
X/Y Force (mN) Load Cell reading
X/Y Displacement Reports the change in size relative to the initial size
Set Name Name specified for each set
Temperature (°C) Current temperature

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Advanced System Settings Dialogue

Advanced settings are available by selecting Advanced from the Settings menu. All of these
settings are template specific.
True Strain Rate
Unlike nominal or engineering strain rate, a plot of
velocity versus time for a true strain rate test would
be non-linear. The degree of non-linearity
increases with increasing strain. The BioTester
approximates this non-linear curve with a series of
linear segments. Typically 10 line segments is a
sufficiently close approximation. More segments
can be used, but you should bear in mind that the
actuator velocity can only be updated every 250ms.
Data Averaging
The load cells are sampled at 100Hz with hardware
using a central moving average of 8 samples.
Further data averaging can be applied to smooth
noisy data.
Force Control Settings
Force Control Settings affect the rate and stability
at which forces are achieved. These settings apply
to preloading as well as Force Control Modes.

Kx and Ky (Default = 100, integer values) are proportional gain tuning parameters (one for
each of the coordinate axes). They control the sensitivity or the amount of corrective action
that is applied to the difference between the current and target force values. At low gain
values the system may be safe and stable but may be sluggish in response to changing
conditions. If the proportional gain is increased the system becomes more responsive and
loads are achieved more quickly. If Kx and Ky are set too high, an unstable oscillating
system may result and forces may never be achieved. The optimal value for these settings
depends on the properties of the material being testing. For stiff tissues values may range
between 1 and 10. For soft tissues values may range between 50 and 200.Kx and Ky usually
are set equal to each other unless testing materials are strongly anisotropic.
Tolerance (mN) (Default = 3) controls how close to the specified force the actual load values
need to be. For example, a specified force of 100 mN with a Tolerance of 3 mN will be
considered achieved when the loads are between 97 and 103 mN. Keep in mind the
tolerance of the load cell being used when choosing the value for this parameter. The load
cells typically fluctuate by about 0.1% of their full scale during the course of a test. For a
1000mN load cell this is about 1mN but for a 23N load cell, this is about 23mN. This value
should usually be 0.2-0.5% of full scale to ensure stable operation.
Settling Frames (Default = 5) controls how many successive load samples (sampled at 100
Hz) need to be within the Tolerance criteria for the force to be considered achieved. For
example, a specified force of 100 mN with a Tolerance of 3 mN and Settling Frames = 5 will
be considered achieved when the loads are between 97 and 103 mN for 5 successive data
samples.
Velocity (1-10 integer values) matches actuator jog speed. Typically for harder tissues, this
value should be between 4 and 6 and for soft tissues, this value should be between 6 and 10.

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Acceleration (1-10 integer values). Typically this should be set to the default value of 10
(maximum acceleration). Changes made to the acceleration will affect tests under both force
and displacement control. Reducing this value may help smooth force controlled tests under
low loads or displacement controlled tests at high speeds. Tests run with lower accelerations
will have softer transitions at direction or speed changes but may take longer than specified
to achieve testing cycles.

BioTester Tip: Testing Force Control Settings

It is important to have appropriate Force Control Settings that match the materials you
are testing.

To safely test Force Control Settings: press on

the tool bar, or from the Tools menu select Move
Actuators to Specified Load. The dialogue shown on
the right will appear. The BioTester will attempt to
achieve the specified loads (forces) but will stop
when either the specified loads are reached, the
actuators have displaced by the maximum specified
amount, or the timeout time has elapsed.
If the actuators oscillate around the specified loads
but never achieve them try reducing Kx and Ky,
and/or decreasing the velocity.

If the actuators reach the specified loads too slowly,

try increasing Kx and Ky, and/or increasing the
velocity.

Soft Limits
To avoid rake collisions or sample destruction, it is useful to assign reasonable travel limits to
the actuators. This is especially important during force control or preloading, where if
specified loads cannot be reached, the actuators will keep moving until they reach the soft
limits.
Min and Max Displacement, in µm, set the minimum and maximum positions to which the
rakes are allowed to move.
Min and Max Force, in mN; this feature is currently not functional. Appropriate limits should
be set using Min and Max Displacement.

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Configuring the Live Charting Graphs

The three graphs on the right side of the
screen provide real-time user feedback during
a test. These graphs are intended to be used
for qualitative feedback, not for detailed
analysis.
The graphs are updated at a maximum
frequency of 10Hz; certain short-duration tests
may not show all the detail that is actually
present.
The graph settings can be specified by
selecting Graphs from the Settings menu. Auto
scaling allows the axis min and max to start at
the specified values but expand if the data
values are larger or smaller than the initial
limits.

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Graph Data
Force versus Time
This graph shows how the measured X and Y forces
are changing with time. Peak loads per cycle,
differences in load between the X and Y axes, and
force relaxation are all easily seen in this type of graph.
Force is proportional to nominal (engineering) stress.

Displacement versus Time

This graph shows how the rakes are moving with time.
The phases of the test sequence and size adjustments
due to preloading are readily apparent in this type of
graph. Displacement is proportional to nominal
(engineering) strain.

Force versus Displacement

This graph displays a qualitative representation of the
material behavior. Viscoelastic effects (like hysteresis)
and material response to different loading phases are
apparent in this graph. This graph is proportional to a
nominal (engineering) stress-strain graph.

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6. Reviewing Test Results

Overview
The BioTester software has an integrated image analysis module which can be accessed by
selecting Analyze and Review Images from the file menu, then selecting the appropriate test file.
The test file is a text file with a .tst extension that contains information about the test parameters
so that the software can display the images and data in a useful fashion.

Image List

Image Playback

Image Tracking

Data Overlay

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Selecting Images
The Images are shown in a tree structure on the top left panel.
The tree structure organizes the images by:
Set (line on the test parameter window)
Cycle (iteration of a given Set)
Phase (Preload/Stretch/Hold/Recovery/Rest of each Cycle)
Image (the one or more individual images from each Phase)
You can select which images to include in a playback set in
one of several ways:
 hold down the shift key and click on individual images
 select entire sets, cycles, or phases (will playback all
contained images)
 select images which have data associated with a
particular tracking set

Image Playback Options

The controls on the left of the screen are broken into 2 sections:
Playback and Tracking. The playback buttons allow the user to review
the images taken during a test.
The playback buttons allow the user to review the images taken during
a test.

Next/Prev Display next or previous image (as dictated by “Play Every” parameter).
Play/Stop Start and stop the playback.
Prev/Next Cycle Jumps to a corresponding image and phase in the next or previous cycle. Useful when reviewing
sets that have multiple cycles.
Loop Sequences can be played continuously in a loop or only once.
Playback Rate +/- changes the speed of the playback. A playback rate can also be manually entered in the
display box.
Play Every Allows the users to skip some frames to expedite playback.

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Image Tracking: Overview

Image tracking is a function that can be used to quantify the
motions of image features (rake tines, specimen texture and
fiducial markers). This can be useful for studying localized
specimen deformations, verifying strain magnitudes, and
comparing the results of one test to another.
The image tracking engine is based on a template matching
algorithm. This algorithm starts by defining a “patch” of pixels
(called a template) surrounding the selected source point on the
source image. It then determines the optimal location for this
template on a target image within a specified search region.
The algorithm defines the location of the tracked point on the target image as the centre of the
optimally located template.
To track an image, follow these steps:
1. Click the Create button in the
tracking window, shown above.
This brings up the Tracking Editor
dialogue box.
2. Enter a Tracking Name.
3. Select the Source Image in the
test image list tree, and click Set
to the left of the Source box.
4. Select the Target Images in the
test image list tree, and click Set
to the left of the Target box. The
target image list will automatically
be sorted to remove duplicates
and be put in sequential order.
5. Generate source points on the
source image by performing the
following steps:
a. Manually click source point
locations on the source image
with the Add button depressed.
b. Draw a box on the source image with the Select button depressed, then click the
Grid button and define a grid of points.
The displayed image will automatically change to the source image when either the Add
or Grid buttons are depressed.

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6. Modify search parameters:

Source The optimal source template is difficult to predict, but in general it is
Template advantageous for the template to be large enough that it contains at
least one feature, but not so large that it contains multiple features.
Typical values for this parameter are between 15 and 55, with 35
being a good starting point for most users.
Search Region The search region needs to be large enough that the optimal
location for the template is found. In general, no part of the
specimen moves more than the rake tines, so the motion of the
rake tines is a good guideline for how large the template needs to
be. For example, if the rake tines move 50 pixels over the 10
frames you are interested in, it is unlikely that any part of the
specimen is moving more than 5 pixels/frame and so a good value
for the search region parameter would be 11 (5 pixels each way,
plus the centre pixel).
Use Source First Image tracks all images against the first in the series (1→2,
Template From 1→3, 1→4…).
Previous Image tracks each subsequent image against the image
before it (1→2, 2→3, 3→4…).
First Image Tracking tends to have poorer correlation than Previous
Image Tracking because the current image is usually closer in
content to the previous image than it is to the first image in the
sequence. Previous Image Tracking tends to have good correlation
for each individual step, but is prone to error accumulation since
each step is independent. The best method for a given data set will
depend on the number of images and the content of the images.
Sometimes it is best to use Previous Image Tracking but to only
th
use every other image or every n image rather than all the images
available. Sometimes it is best to use First Image Tracking and use
every image but set the source image in the middle of the image
series rather than at the beginning.
Set Search New Point Locations The location of the centre of the search
Region Centre region is set to the same location of that point in the previous image
(even when the source template is from the first image).
Existing Point Locations The search region centre location is set
to the previously tracked location of that point. When no previously
tracked point exists, the location is chosen in the same way as New
Point Locations.
Use Strain When this option is checked, the source template will be scaled
Based (resized) according to strains calculated from the test XSize and
Deformable YSize parameters. Best tracking results are often obtained when
Templates using this option. This option may not be the best for uniaxial tests
or tests where the specimen deformation is non-uniform.
7. Perform tracking with either Track All Points or Track Selected Points.

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Image Tracking: The Points Display Option

The Points Option displays the points in their current
location (i.e. their location on the currently displayed
image) with or without gridline connecting points that
were generated using the grid function.
Clicking the details icon brings up the window shown
to the right. Options include changing the point radius
or color, showing ID numbers and showing gridlines
with or without ID numbers.
Gridline ID numbers are associated with each grid
square (cell), while Point ID numbers are associated
with the points themselves.
Tracking display settings default to the last used
settings.

Figure 1: Initial tracking grid displayed as Figure 2: Tracking to 20% strain displayed as
points. points.

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Image Tracking: The Displacement Option

The Displacement Option displays the points in their
current location as well as graphically representing their
motion through time.
Clicking the details icon brings up the window shown to
the right. This dialogue allows the user to change the
reference image, change the type of connecting line,
and change the line appearance.
Displacement connecting lines directly connect the
reference and current points via a vector or a path. The
path option provides more information but can result in
slow playback speeds for sets with many points and/or
many images.
The Flow option generates a direction vector based at
the current location and oriented in the direction of
motion relative to the reference image. This can be
useful because the length of this vector can be scaled
to better visualize small displacements.
Tracking displacement settings default to the last used
settings.

Figure 3: Tracking to 20% strain displayed as displacement.

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Image Tracking: The Strains Option

The Strains Option can only be used in conjunction with
a grid of points (as opposed to individually placed
points). This option calculates the regional strains
inside every grid box and displays this data as an
ellipse. One can imagine this is what would happen to a
grid of circles drawn on the surface of the specimen at
the beginning of the test.
Clicking the details icon brings up the window shown to
the right. This dialogue allows the user to change the
reference image as well as the appearance of the
circles and text.
Showing text for E1, E2 will result in the text showing
the strains along the major and minor axis of the
ellipse. Showing text for Ex, Ey will result in text
showing the strains along the X and Y directions
regardless of the ellipse orientation.
Tracking strain settings default to the last used settings.

Figure 4: Tracking to 20% strain displayed as Figure 5: Tracking to 20% strain with text
strain ellipses. showing x% strain.

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Data Overlay
Data associated with each image is displayed below
the tracking controls on the left side of the main
window. Checking the Show Graph box displays this
data as a graph overlaid on the images

Clicking the details icon displays the window shown to

the right. The dialogue allows the user to change the
size and appearance of the overlaid graph. The data
points associated with the current image are marked on
the graph with green circles. The overlaid data graph
will be drawn on top of any image tracking displays
(when they overlap).
Data Overlay settings default to the last used settings.

Figure 6: Tracking to 20% strain with overlaid x-y force graph.

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Exporting Tracked Data

Tracked grid point coordinates and calculated grid
strains can be exported to a .csv file. From the Export
menu, select Data Points to display the dialogue shown
on the right. The image list is automatically set to the
currently selected images in the test image list tree.
This image list can be changed by selecting new
images in the test image list tree and pressing the set
button. When exporting Strains data from Selected
Tracking Points, all four points of a grid cell need to be
selected for data from that cell to be exported.
Exported data is labeled with associated ID numbers
(grid point ID numbers for exported coordinates and
grid cell ID numbers for exported strain values). To
display ID numbers on tracking grids see Image
Tracking: The Points Display Option. In the following
example (Figure 9), grid point ID numbers are shown in
yellow and grid cell ID numbers are shown in white.
When exporting data for the selected points as shown
in red, coordinates are exported for points with ID
numbers 5, 6, 7, 8, 9, 10, 11 and 12, and strains are
exported for grid cells with ID numbers 4, 5, and 6.

Figure 7: Tracking Grid showing ID Numbers.

Figure 8: Sample Exported Data.

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Exporting Images and Movies

The current Image view, including all displayed tracking
grids, tracking data and overlaid data can be exported
as a metafile by selecting Current Image from the
Export menu. Alternately, the current image view can
be copied to the clipboard by selecting Ctrl-C from the
keyboard.
To export a series of image views, including all
displayed tracking grids, tracking data and overlaid
data as an .avi movie, select Movie from the Export
menu. The dialogue shown on the right will be
displayed. The image list is automatically set to the
currently selected images in the test image list tree.
This image list can be changed by selecting new
images in the test image list tree and pressing the set
button. Options are available for setting the fame rate
or image size (resolution), adding filenames, and
choosing color or grayscale output. Compression
options are also available.

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7. System Hardware Settings

Controller
The BioTester uses an integrated control board rather
than a group of PC controlled device drivers. This
minimizes communication lag and allows the device to
operate asynchronously with the PC. The control board
details can be seen by selecting Hardware from the
Settings menu, which brings up the window shown to
the right.

Camera
The camera uses a ½” monochrome CCD sensor to
obtain a resolution of 1280 pixels by 960 pixels. The
camera shutter can be controlled during the setup
phase of the test.
The camera captures images at 15 frames per second
during every test and writes these images to a buffer.
The test controller extracts images from this buffer at
the frequency specified in the Set Parameter Editor
(15Hz, 5Hz, 1Hz, 0.1Hz, 0.01Hz, or 0.001Hz). These
images are written to the output directory. The file
name is the ideal capture time. The actual capture time
may differ from the ideal capture time by as much as
1/30 of a second since the images are extracted from a
buffer.
The test software creates an image capture log file in the output directory should this difference
be significant. This log stores the actual capture time and the idealized capture time.
The camera default shutter and gain are specific to each test template and can be set by
selecting Hardware from the Settings menu. Typically the gain is best left at 20 and the shutter
changed to alter the brightness of the image.

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Load Cells
The load cells are semiconductor strain gage-based
with mechanical overload protection. They have
accuracy equal to 0.2% of the rated full scale load. The
load cells are available in 0.5N, 1N, 2.5N, 5N, 10N and
23N sizes. Each load cell has a unique calibration
factor which is stored in a chip attached to the load cell
connector.
While the load cells should not have significant
hysteresis or thermal drift, temperature changes or
prolonged loading can introduce small offsets. It is
recommended that the zero load state be redefined
periodically. This can be done by selecting Zero Load
Cells from the Tools menu or pressing .
The load cells are sampled at 100Hz, with hardware sample averaging to reduce noise in the
data. Further sample averaging can be implemented by selecting Advanced from the Settings
menu.
Each load cell is calibrated and has a specific calibration factor which is stored on a chip in the
load cell connector. The range and calibration factor can be viewed by selecting Hardware from
the Settings menu, but are not user controlled. In addition, the load cells need to be calibrated
when the system is initially set up and whenever load cells are changed or re-mounted. See
Section 8 for this procedure.

Actuators and Motors

The actuators are driven by a stepper motor under
open loop control. The motor drives a lead screw
attached to a stage that the gooseneck is mounted to.
All 4 axes are driven independently. The stages have a
peak velocity of 5mm/s. Provided that the actuators are
not driven beyond their rated velocities and thrust
loads, the stepper motors are very unlikely to lose
steps. To prevent excessive velocities and
accelerations, the control software will issue warnings if
entries in the Set Parameter Editor result in
accelerations or velocities that are out of range.
Excessive thrust loads should only occur if mechanical
interference from the fluid chamber or a foreign object
obstructs the movement of one or more actuators. It is
advisable to reset the actuators occasionally.
Idle current (holding current) can be applied to the motors. Since the actuators are self-locking
(they cannot be moved once power is removed) the default is to have the motors de-energized
when they are not moving. Turning idle current on may produce smoother motions in force
control (since the motors are frequently stopping and starting). When idle current is activated the
motors will produce more noise.

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Temperature
To activate the fluid chamber heater, both power
switches on the front of the BioTester need to be in the
On position (the computer and software do not need to
be on yet). At this point, the heater will warm the fluid to
the last used temperature set point. The temperature
set point can be changed by selecting Hardware from
the Settings menu.
Typically, the heater will take 20 to 40 minutes to warm
a full fluid chamber from room temperature to 37°C.
When Show Temperature Warnings is enabled and the
Current Temperature is not at the Temperature Set
Point plus or minus the specified tolerance -- a red
indicator button will be displayed next to the Current
Temperature live output and a warning dialogue will
appear when you execute the test.

System Alert
In the event of a dislodged or faulty temperature sensor, or if there is a lack of fluid in
the fluid chamber, the system is designed to reach a maximum temperature of 80°C.

Caution: Avoid touching the specimen stage when the heater is on, it can become
very hot!

BioTester Tip: Preheat Fluid

You can add preheated fluid to the fluid chamber to reduce warm up time.

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External Sync Pulse

A 5 volt TTL external sync pulse signal can be
generated on BioTester models containing a BNC
connector jack on the back panel.
The sync pulse will be pulsed on the very first sample
pulse, which occurs when the load cells are measured
and the motor positions of all 4 axes are read.
Thereafter a sync pulse will occur at a user definable
frequency as fast as 100 Hz (1 pulse every 10 mS) and
as slow as 0.00305 Hz (1 pulse every 327 s). The sync
pulse will stop when the sampling has been stopped.

The sync pulse is active high for 10 mS except when

operating at maximum speed (100 Hz) where the active
high pulse duration is about 550 nS.

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8. System Calibration and Advanced Tools

Snap Image Feature
Located under >Tools>Snap Image
This feature allows the user to capture a single image during the setup phase. This can be useful
for documenting specimen loading activity and neutral (non-contact) beam position.

Load Cell Calibration

If the load cells are changed, moved, or impacted, it is good practice to perform the load cell
calibration routine. This calibration can be performed in the following steps:
1. Launch the calibration utility by selecting Tools, Advanced, then Load Cell Calibration
2. Input the K value (spring constant) for the calibration spring provided for the load cell
being used (shown on the spring case label, provided upon system delivery)
3. The load cell capacity should be automatically read and displayed, and the appropriate
calibration preload calculated
4. Lower the fluid chamber and install the calibration spring brackets.
5. Zero the load cell using the Zero Load Cells button
6. Using the Jog+ and Jog– buttons, drive the actuators until the posts are correctly spaced
so that the calibration spring can be installed without applying any significant load. Set
the calibration spring on the posts (see above figure)
7. Using the Jog+ and Jog– buttons, move the actuators until the spring is carrying a
tension that is out of the non-linear “dead zone” and reaches the appropriate preload
displayed in the Calibration Preload dialogue. For most setups, the spring should be
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driven to 30% of the load cell rating ie. for a 2.5N load cell, a preload of 750mN is
specified. If it is suspected that the load cell is more than 15% out of calibration, contact
the manufacturer for another recommendation for calibration pre-load.
8. Visually verify that the spring is loaded in a stable, horizontal position.
9. Click the Run button, wait for calibration to complete.

10. The number (A) that is displayed in the main dialogue box
at the end of the calibration procedure is the ratio of the
current calibration value to the previous calibration value.
If there has been no change to the load cell, A should be
between 0.99 and 1.01.
11. If the A value is greater than 1.05 or less than 0.95, the
load cells were significantly out of calibration. If this is the
case, the calibration procedure above should be repeated
to ensure that the preload was correct and an accurate
calibration achieved.

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Adjusting the Camera Position and Image Magnification

The camera position can be adjusted using the knobs at the back and side of the camera
housing. If larger position adjustments are needed, the bolts that attach the camera to the mast
can be loosened to allow rotation of the camera housing.
The BioTester comes equipped with a 75mm focal length lens and a set of lens spacers. Using
these products it is possible to achieve a field of view ranging from 9mm to 30mm. The details of
the appropriate number of spacers and the required standoff are shown in the table below. The
image magnification can be changed by adding or removing spacer tubes located between the
camera body and the lens. Increasing the total spacer tube length will result in higher
magnification while reducing the tube length will result in lower magnification. Changing the tube
length will also change the working distance of the lens. To adjust the vertical position of the lens
and camera assembly, loosen the bolts that clamp the camera housing to the mast and raise or
lower the camera housing until the image is in focus. It is helpful to make sure the lens focus ring
is in the middle of its rotation so that fine tuning the image focus can be done once the camera
housing is re-clamped.
INCREASING THE TOTAL LENGTH OF THE SPACER TUBES WILL INCREASE
MAGNIFICATION (DECREASE FIELD OF VIEW) AND DECREASE WORKING DISTANCE
(STAND-OFF).
If the camera position or magnification is changed, it is important to capture a single image scale
shot. A glass calibration target or a ruler is acceptable for this purpose. The scale shot will be
useful later to characterize the size of test specimens or test specimen features.

As the table and graph also show, it is also possible to swap in a lens with a different focal length
to achieve a different range of magnifications. This data was collected using the family of fixed
focal length lenses available from Edmund Optics (the part numbers are shown in the table). The
camera has a C-mount connection, making it possible to mount a wide variety of optical
components to best suit any application. Currently, only the standard camera is supported.

25mm (NT56-529) 35mm (NT56-530) 50mm (NT56-531) 75mm (NT56-532) 100mm (NT56-675)
Stand Stand Stand Stand Stand
Spacer off pix/c FOV off pix/c FOV off pix/c FOV off pix/c FOV off pix/c FOV
(mm) (mm) m (mm) (mm) m (mm) (mm) m (mm) (mm) m (mm) (mm) m (mm)
0 685 78.6 162.8 1070 71 179.3 930 121 105.9
5 100 520 24.6 215 373 34.3 370 329 38.9 820 206 62.2 1540 147 86.8
10 53 935 13.7 125 682 18.8 235 560 22.9 450 404 31.7 950 258 49.6
15 90 993 12.9 185 779 16.4 350 534 24.0 675 366 34.9
20 73 1273 10.1 160 978 13.1 285 711 18.0 535 473 27.0
25 135 1186 10.8 245 859 14.9 455 578 22.1
30 125 1410 9.1 220 959 13.3 400 687 18.6
35 115 1630 7.9 200 1111 11.5 355 797 16.1
40 105 1842 6.9 185 1260 10.2 320 897 14.3
45 100 2068 6.2 170 1392 9.2 295 1003 12.8

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600

500

400
Standoff (mm)

25mm

300 35mm

50mm
200 75mm

100mm
100

0
0 20 40 60 80 100 120 140
Field of View (mm)

NOTE: The physical camera standoff will not go below 100mm or exceed 500mm, the preceding
table and chart provides guidelines only

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Update Firmware

1. With the BioTester connected to the PC and turned on, launch the firmware update
software located in the Windows start menu under LabJoy>Utilities.
2. Load the firmware file using the “Load Update File” button.
3. Click the “Connect” button.
4. Execute the update using the “Update” button and wait for update to complete. This
can take many minutes.

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9. Troubleshooting

Image Appears Out of Focus

With the live image on the screen, adjust the lens focus until the image appears to be in focus.
Whenever you adjust the focus, the image magnification may change slightly. If you want to know
the absolute scale of your Images you can snap an image of a calibrated target (such as a
precision graticule or ruler) by selecting Snap Image from the Tools menu. You can estimate the
image scale by comparing the number of pixels between rake tips in the images with their known
µm spacing (size) in the data output files.

ERROR - Sample Buffer Overflow Continuing Test...

or
System Lagging, Camera Frame Lost missed capture
These errors can be caused by lack of data throughput to the location where the test data is
being saved. This has been noted by users who are saving their data to a network drive or a
USB-connected external hard drive. Another cause for these errors can be a lack of computer
resources, especially at high image capture frequencies (5 or 15Hz). It may be necessary to
upgrade the computer that is controlling the device.

Communication Errors: ReadFile failed with error 6.

When communication errors occur the live message window will display a ReadFile failed
message. Communication errors can occur if:
 The BioTester is not powered up.
 USB cables between the computer and BioTester are disconnected.
 A USB 3.0 port is used
 The supply power surges or is interrupted.
When a communication error occurs, close LabJoy, check connections, power cycle the
BioTester, and restart LabJoy.

BioTester Tip: Preventing Power Disruptions

We recommend plugging the BioTester (and host computer) into an uninterruptable
power supply (UPS) or battery back-up.

Zero Displacement Causes Software Crash

Under certain conditions involving preloads and cyclic loading, the software may crash if a test
phase has zero displacement in one axis. If this happens, the easiest solution is to replace the
zero displacement with a small non-zero displacement value (i.e. 0.1% or 10μm).

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10. Appendix A: Initial System Setup

The BioTester has been packed in a special container to protect it during shipping. In addition,
system accessories have been packed in several cartons contained in a separate box. The
system components listed below may be unpacked and laid out before system setup. A space
24” x 24” with 40” overhead clearance is required for setup.

Tools

Small flat head screwdriver

SAE hex wrenches: 1/16” 3/32” 7/64” 9/64”
SAE Combination Wrench 5/16”

Fasteners & Parts

1 BioTester
16 6-32 X 1/2” Socket Head Cap Screws
2 6-32 Hex Nuts
12 5mm Magnets
12 7.5mm Magnets
20 BioRakes
1 Calibration Spring
2 Calibration Spring Brackets
1 Set of Camera Lens Spacer Tubes
1 Specimen Mounting Bridge
2 Load Cell Goosenecks
2 No-Load Cell Goosenecks
6 6-32 Hex Nuts (2 extra)
1 Standard Specimen Cutting Jig
4 Razor Blades for Cutting Jig
1 Fluid Bath Chamber
1 Power Cord and Supply
1 6’ (1.8m) USB Cable
1 Set of Spacer Plates
1 Sheet of Rubber Backing Material
2 Load Cells

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Unpack System

Remove the BioTester from the shipping container. Carefully remove the boxes and protective
shipping material from the container such that the BioTester can be lifted out. This will require 2
people. Take special care with the camera mast as it is connected to the main unit by cables. In
addition, protective foam will have to be removed from the main unit and camera mast.

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Attach Camera Mast

Thread the mast into the receiver. The knurled collar must be rotated to fasten the camera mast.
It will take many turns but eventually the mast will lock into place. Try to position the camera
approximately as shown, although the final position can be adjusted later in the procedure. Be
sure to reduce the slack on the cables before threading the post, so as not to pinch the cables
while tightening the collar.

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Attach Magnets to Goosenecks

Check the orientation by attempting to place a BioRake in the gooseneck. If the magnets are
correct, then the BioRake will snap in to place. If the BioRake is repelled by the magnet, then
they have been installed backwards.

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Place Fluid Chamber on Riser Stage

Align the temperature probe mounting hole with temperature probe cable.

Install Camera Lens

Remove protective caps on the lens and camera body and then screw the lens into the camera
body. Spacer tubes are placed between the lens and the camera body to change the image
scale. This also affects the working distance of the lens and therefore the camera housing may
need to be moved up or down on the camera mast to accommodate changes in the number of
spacer tubes.

Lens (typ. 75mm) Spacer tubes Camera (inside housing)

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11. Appendix B: Software Installation

A Windows based PC with at least 2 USB 2.0 ports is required to operate the BioTester. Included
in your shipment is a USB key which contains the LabJoy software. These files contain the
software installer for LabJoy, which is the user interface and control software for the BioTester.
To install the software, simply insert the USB key, open the flash drive, and click on “setup.exe”.
Details of typical dialogue boxes are shown below.

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12. Appendix C: Install Load Cells / Perform

Camera and Gooseneck Alignment

Connect USB cables to the PC USB 2.0 ports and then proceed
with the following steps.

Attach the Load Cells to the Goosenecks

Ensure the correct orientation of the load cells. Tighten the nuts gently to avoid damaging the
load cells (just a little bit tighter than finger tight). Keep in mind that the load cells are sensitive to
excessive torque, excessive overload, transverse loading, and bending forces. Ensure the load
cell cable angle is approximately as shown (see following 4 images).

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Attach the Goosenecks to the Actuators

See the following 4 images. The nut on the backside of the load cell is not as sensitive as the
front side nut. Make sure that the gooseneck is rotated such that it is level when fastening the
backside nut.

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Plug in the Load Cells

After connecting the load cells, tuck the cables inside the plastic cover, taking care to handle the
connectors by the edges. A circuit board assembly may be attached to the connector and must
be handled carefully. Position the cables inside the cover so that they do not interfere with the
actuator carriage travel, particularly during actuator reset where the carriages are fully retracted.

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Launch the LabJoy Software

Launch the LabJoy software. The default screen should appear as:

Selecting File>Collect New should bring up this window:

The details of the software are outlined in this user’s manual. For assembly purposes, it is only
required that we initiate the default test. Select OK.
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If prompted to perform an actuator reset, select “yes”.

If prompted to perform a load cell calibration, select “no”.

Next place a BioRake in each of the 4 goosenecks so that the BioTester looks like:

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On the LabJoy control screen, ensure that the specimen size is set to 5000µm in X and Y and
then click the “Move to Size” button.

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Perform Camera Alignment

The camera position has both coarse and fine position adjustments and coarse and fine focus
adjustments. To start, make sure the lens fine focus knob (lowest ring on lens) is approximately
in the middle of its rotation. Next, loosen the bolts that clamp the camera housing to the mast.
The camera assembly can then be rotated until the camera is positioned approximately over the
centre of the 4 BioRakes. As well as rotating, the camera assembly can slide up and down to
bring the BioRakes into approximate focus. Be careful to move the camera housing gently in
small increments so as not to crash it into the goosenecks or BioRakes. Attempt to achieve the
setup approximately as shown below:

Next, fine tune the camera position using the knobs at the back and side of the camera housing
and fine tune the image focus using the lowest ring on the lens.
Adjustments to the image scale can be made by changing the overall height of the spacer tubes
between the lens and the camera. See section on “Adjusting the Camera Position and Image
Magnification” (page 37).

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Perform Vertical Gooseneck Alignment

Because the goosenecks attached to the load cells are connected by only 1 threaded stud, the
vertical and transverse alignment of the end of the gooseneck can be somewhat different each
time the goosenecks are removed and reattached. To accommodate the vertical variation, shims
between the gooseneck brackets and the actuators may need to be added or removed. Thick
and thin shims are provided (3 thins shims = 1 thick shim). At this stage, do not be concerned
with the transverse or axial positioning of the goosenecks as these will be aligned in subsequent
steps.

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By looking at the BioRakes from the side, it is possible to observe their vertical alignment. Please
add or remove shims to vertically align the BioRakes. Here are pictures of poor vertical alignment
and good vertical alignment:

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Perform Transverse and Axial Gooseneck Alignment

This step requires 2 different adjustment mechanisms. The goal is to make the BioRakes look
like this:

First, make sure the BioRakes are positioned in the transverse centre of the gooseneck. The
magnetic mounts that hold the BioRakes to the gooseneck fix them in the axial direction, but
allow a small amount of adjustment in the transverse direction. This is useful for alignment during
regular testing, but for the purposes of alignment they should be in the neutral position. The first
image below shows a transversely offset BioRake and the second shows a transversely centered
BioRake.

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To transversely adjust the goosenecks, loosen the bolts that attach the goosenecks to the
actuators, slightly rotate the gooseneck, and retighten the bolts. The on-screen image should be
helpful as a guide.
To axially adjust the goosenecks, use the actuator jog icons in the lower left of the screen. Make
sure the “independent” radio button is selected and not the “mirror matching” button. The jog
speed should be about 7.

Adjust all 4 goosenecks until you are satisfied with their transverse and axial alignment. It may
also be necessary to make small adjustments to the camera position.
It is now necessary to perform a centre position calibration. First, it will be necessary to
temporarily define the current position as the centre position. This calibration can be performed
in the following steps:
1. Lower the fluid chamber and remove the BioRakes.
2. From the Tools menu select Advanced and then select Centre Position Calibration. This
should bring up this dialogue box:

3. Press YES.

4. Select the reset actuators button (or use >Tools>Reset Actuators).

5. From the Tools menu select Advanced then select Move to Centre. This should bring up
this dialogue box:

6. Press YES. Once the actuators have stopped moving, insert 2 opposing BioRakes on
the X axis.
7. Using the actuator jog control arrows at the lower left of the screen, move the actuators
until all 10 BioRake tips are collinear. Be sure to jog the BioRakes in MIRROR
MATCHING MODE ONLY!!! Note that it is the tips of the BioRakes that should be
collinear and not the outside edge. Because the tips are underneath the BioRakes and
cannot be seen on screen, it is necessary to estimate their position. Please use the
image below as a reference:

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Example 1: BioRakes have too much overlap.

Example 2: BioRakes are correctly positioned.

Example 3: BioRakes do not have any overlap so tips are not collinear.
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8. Remove the BioRakes and place them on the other axis.

9. Using the actuator control arrows: Drive the actuators until all 10 BioRake tips are
collinear as in step 4 (IN MIRROR MATCHING MODE ONLY!!!).
10. From the Tools menu select Advanced and select Centre Position Calibration. This
should bring up this dialogue box:

11. Press YES.

12. Select the reset actuators button (or use >Tools>Reset Actuators).

To check the centre position calibration, use the >Tools>Advanced>Move to Centre command
while having only 2 opposing BioRakes mounted. Compare the position of the rakes to the image
in step 4 above.

Install Actuator Covers

Each actuator cover has a number written on the inside. These numbers match up with numbers
written on the inside of the main cover to ensure the best possible fit between the actuator covers
and the main cover.

The best way to install the covers is to place the back 2 pegs into the main cover and then lower
the front 2 pegs while squeezing them together. Be careful not to let the cover slip and
accidentally apply a transverse load to the gooseneck.

Finally, perform load cell calibration (see Section 8 under Load Cell
Calibration).

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13. Appendix D: Protocol for Tying and

Cutting Sutures for Biaxial Testing

There are many ways to attach specimens to the suture brackets using a variety of suture
designs. This document outlines one way to use double-armed sutures to accomplish specimen
attachment.

1. Assemble equipment: small vice, magnifying goggles, 2 needle nose pliers, small forceps.

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2. Bend suture hooks

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3. Tie off sutures. First hook each bent hook around a secured post held in the vice. Then
tie a standard knot in the suture and use forceps to guide the location of the knot so
that it tightens at the correct location.

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4. Use pliers to tighten knot

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5. Cut off excess.

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The resulting mounted specimen should ideally look something like this:

© Copyright 2014 CellScale Biomaterials Testing.

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