Download StageFlexer® User`s Manual - Flexcell International Corp.

StageFlexer®
User's Manual
12-22-14
Rev: 3.0
Culturing Cells in a Mechanically Active Environment™
Flexcell International Corporation  2730 Tucker Street, Suite 200  Burlington, NC 27215
800-728-3714  (919) 732-1591  FAX: (919) 732-5196  www.flexcellint.com
COPYRIGHT © 2009 FLEXCELL INTERNATIONAL CORPORATION
FLEXCELL® INTERNATIONAL CORPORATION
TABLE OF CONTENTS
Using the StageFlexer® ...................................................................................................... 1
Introduction ........................................................................................................................ 1
StageFlexer® Assembly ..................................................................................................... 1
Plating and Viewing Cells when using a Loading Station™ .............................................. 3
Using the StageFlexer with the FX-5000™ or Flex Jr.™ Tension System ......................... 3
Minimum Loading Station™ Limitation ............................................................................ 3
Appendices ........................................................................................................................... 4
Appendix 1: StageFlexer® 25 mm Loading Station™ Conversion Chart .......................... 5
Appendix 2: StageFlexer ® 28 mm Loading Station™ Conversion Chart ......................... 6
Appendix 3: StageFlexer ® 31 mm Loading Station™ Conversion Chart ......................... 7
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FLEXCELL® INTERNATIONAL CORPORATION
USING THE STAGEFLEXER®
INTRODUCTION
The Flexcell® StageFlexer® (Fig. 1) in
conjunction with the FX-5000™ or Flex Jr. ™
Tension System allows users to observe cell
stretching activity under a microscope. Cells
are grown on a 42 mm diameter silicone
membrane (StageFlexer® Membrane) which
is clamped and sealed above a small
cylindrical vacuum chamber. When a
vacuum is applied, the membrane is pulled
downward into this chamber applying strain
to the cells growing on the membrane
surface (Fig. 2). The membrane can be
deformed freely in an open chamber or over
a loading post to supply uniform biaxial
strain to the cells (Fig. 2). The loading post
comes in three different diameters (25 mm,
28 mm, and 31 mm) for varied growth
surface area.
Figure 2. Schematic of strain application to cells in a
StageFlexer®
STAGEFLEXER® ASSEMBLY
Assemble the eight main parts of the
StageFlexer® in the following order (Fig. 3):
1. StageFlexer® body
2. Rubber gasket
3. Loading Station™
4. Retaining ring
5. O-ring
6. Silicone membrane
7. Top ring
8. Four top screws
Figure 1. Flexcell® StageFlexer®
Cells in a StageFlexer® can be viewed on a
standard microscope. The hole in the bottom
of the StageFlexer® allows for illumination
through the loading post.
Figure 3. StageFlexer® assembly order
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FLEXCELL® INTERNATIONAL CORPORATION
The Loading Station™ (i.e., loading post) is
a small hollow acrylic piece that is inserted
into the StageFlexer® vacuum chamber. It is
used with a rubber gasket and a retaining
ring that effectively seal the vacuum within
the unit.
layer between the silicone membrane and
Loading Station™ to reduce friction.
Next, place the O-ring over the ridge around
the main body of the StageFlexer®. Then
carefully place the membrane over top of the
vacuum chamber and O-ring and center it
using the screw holes around the edge of the
StageFlexer®. Finally place the top ring over
the membrane and O-ring, put the screws in
place, and tighten the assembly to clamp the
membrane.
To insert the Loading Station™ into the
StageFlexer®:
1. Coat the rubber gasket with a thin film
of silicone grease on both sides and
insert it into the bottom of the
StageFlexer®.
Add medium to the cells as needed.
Connect the 1/4" blue tubing with BioFlex®
adapter to the fitting on the front of the
StageFlexer®. The base of the assembly
should fit onto the microscope base for
proper viewing.
2. When using the 25 mm Loading
Station™, place it into the StageFlexer
first. Using retaining ring pliers to
squeeze the retaining ring, close the ring
enough to push it into the StageFlexer®.
Push the ring down until it snaps into the
milled ring hole at the bottom of the
StageFlexer®. Getting the ring low
enough may require additional vertical
pressure from a regular screwdriver or
similar tool.
Also included with the StageFlexer® is a
larger aluminum plate which can be screwed
onto the microscope stage. This plate
replaces the small plate that is screwed to
the bottom of the StageFlexer®. Simply
remove the four screws on the bottom of the
StageFlexer® and then the small plate.
Screw the larger plate in its place such that
the long end is facing away from the fitting
on the front of the StageFlexer®. The larger
base will have to be drilled at the back end
to fit onto your particular microscope. The
stage clips on your microscope should be
held in place by two screws at the back of
the stage. The holes to be drilled in the back
of the larger base should be distanced apart
exactly to fit the two screws that hold the
stage clips in place. Once the holes are
dimensioned and drilled in the larger
StageFlexer® base, attach the base to the
stage of the microscope with the screws.
This will allow you to easily adjust the
position of the StageFlexer® on your
microscope and keep it in place over time.
3. When using the 28 mm or 31 mm
Loading Stations™, use the retaining ring
tool to squeeze the retaining ring around
the bottom of the Loading Station™ into
the milled ring area. Place the assembly
into the StageFlexer® and push the ring
down until it snaps into the milled ring
hole at the bottom. As before, this may
require the use of a regular screwdriver
or similar tool.
To remove the 25 mm, 28 mm and 31 mm
Loading Stations™, reverse the above
process.
After assembly, use the Lubri-Sil™ lubricant
supplied with the post to spread a generous,
even layer of lubricant on top of the Loading
Station™. The idea is to create a boundary
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FLEXCELL® INTERNATIONAL CORPORATION
PLATING AND VIEWING CELLS
™
USING A LOADING STATION
WHEN
It should be noted that the only cells that are
going to receive uniform strain are those
attached to area of the membrane that is over
the post when the membrane is in its fully
stretched position. Therefore, it is best to
attempt to plate cells only in the uniformly
strained area or to view or test the cells that
are in the uniformly strained area. To
determine this area, the following equation
can be used:
To FX-5000
Diameter = (Diameter of Loading
Station)/(1 + (Max%Elongation/100))
To StageFlexer
Figure 4. StageFlexer® to FX-5000™ Tension Unit
Tubing Adapter
The "Max%Elongation" is the maximum %
elongation that you plan to use in your
regimen. The calculated diameter is the
diameter of a circle at the center of the
membrane. Any cells outside of this circle
will not receive uniform strain.
Also included with the system is a white
PVC volume that should be used inline with
the FLEX IN and FLEX OUT tubing.
Because the StageFlexer® has such a low air
volume, it is necessary that some extra
volume be added to the system for
stabilization. The PVC volume is simply
added by cutting the larger clear tubing on
the FX-5000™/ Flex Jr.™ tubing adapter at
its midpoint, and connecting each cut end to
the fitting on each end of the PVC volume.
USING THE STAGEFLEXER® WITH THE FX5000™ OR FLEX JR. ™ TENSION SYSTEM
Connect the front hose barb of the
StageFlexer® to the FX-5000™ or Flex Jr.™
as shown in Figure 4. Connect the short blue
tubing provided with the StageFlexer® to the
front hose barb on the StageFlexer®. Then
connect the quick disconnect at the end of
the StageFlexer® to the quick disconnect at
the end of the tubing shown below.
MINIMUM
LIMITATION
LOADING
STATION™
It is important to note that there is a
minimum vacuum level in which static
friction is overcome on the Loading
Stations™. The membrane does not begin to
stretch until this point. For the 25mm
Loading Station™, this level is -6.0 kPa or
1.6% elongation. For the 28mm Loading
Station, this level is -12.0 kPa or 1.9%. For
the 31mm Loading Station™, this level is 16.0 kPa or 2.1%. Do not try stretching cells
at a % elongation less than that listed for
each Loading Station™ diameter.
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FLEXCELL® INTERNATIONAL CORPORATION
APPENDICES
Appendix 1:
StageFlexer® 25 mm Loading Station™ Conversion Chart
Appendix 2:
StageFlexer ® 28 mm Loading Station™ Conversion Chart
Appendix 3:
StageFlexer ® 31 mm Loading Station™ Conversion Chart
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FLEXCELL® INTERNATIONAL CORPORATION
APPENDIX 1: STAGEFLEXER® 25 MM LOADING STATION™ CONVERSION CHART
Press (-kPa)
% Elong
Strain
Press (-kPa)
% Elong
Strain
Press (-kPa)
% Elong
Strain
0.00
0.28
0.57
0.86
1.16
1.46
1.77
2.09
2.41
2.74
3.08
3.42
3.76
4.11
4.47
4.83
5.20
5.57
5.95
6.33
6.72
7.12
7.52
7.93
8.34
8.75
9.18
9.60
10.04
10.47
10.92
11.36
11.82
12.27
12.74
13.21
13.68
14.16
14.64
15.13
15.62
16.12
16.62
17.13
17.64
18.15
18.67
19.20
19.73
20.26
20.80
21.35
21.90
22.45
0.0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0.9
1.0
1.1
1.2
1.3
1.4
1.5
1.6
1.7
1.8
1.9
2.0
2.1
2.2
2.3
2.4
2.5
2.6
2.7
2.8
2.9
3.0
3.1
3.2
3.3
3.4
3.5
3.6
3.7
3.8
3.9
4.0
4.1
4.2
4.3
4.4
4.5
4.6
4.7
4.8
4.9
5.0
5.1
5.2
5.3
0.000
0.001
0.002
0.003
0.004
0.005
0.006
0.007
0.008
0.009
0.010
0.011
0.012
0.013
0.014
0.015
0.016
0.017
0.018
0.019
0.020
0.021
0.022
0.023
0.024
0.025
0.026
0.027
0.028
0.029
0.030
0.031
0.032
0.033
0.034
0.035
0.036
0.037
0.038
0.039
0.040
0.041
0.042
0.043
0.044
0.045
0.046
0.047
0.048
0.049
0.050
0.051
0.052
0.053
23.01
23.57
24.13
24.70
25.28
25.86
26.44
27.03
27.62
28.21
28.81
29.41
30.02
30.63
31.25
31.87
32.49
33.12
33.75
34.38
35.02
35.67
36.31
36.96
37.62
38.27
38.93
39.60
40.27
40.94
41.61
42.29
42.97
43.66
44.35
45.04
45.73
46.43
47.13
47.84
48.54
49.26
49.97
50.69
51.41
52.13
52.86
53.59
54.32
55.05
55.79
56.53
57.28
58.02
5.4
5.5
5.6
5.7
5.8
5.9
6.0
6.1
6.2
6.3
6.4
6.5
6.6
6.7
6.8
6.9
7.0
7.1
7.2
7.3
7.4
7.5
7.6
7.7
7.8
7.9
8.0
8.1
8.2
8.3
8.4
8.5
8.6
8.7
8.8
8.9
9.0
9.1
9.2
9.3
9.4
9.5
9.6
9.7
9.8
9.9
10.0
10.1
10.2
10.3
10.4
10.5
10.6
10.7
0.054
0.055
0.056
0.057
0.058
0.059
0.060
0.061
0.062
0.063
0.064
0.065
0.066
0.067
0.068
0.069
0.070
0.071
0.072
0.073
0.074
0.075
0.076
0.077
0.078
0.079
0.080
0.081
0.082
0.083
0.084
0.085
0.086
0.087
0.088
0.089
0.090
0.091
0.092
0.093
0.094
0.095
0.096
0.097
0.098
0.099
0.100
0.101
0.102
0.103
0.104
0.105
0.106
0.107
58.77
59.52
60.28
61.03
61.79
62.56
63.32
64.09
64.86
65.63
66.41
67.18
67.96
68.74
69.53
70.31
71.10
71.89
72.69
73.48
74.28
75.08
75.88
76.68
77.49
78.30
79.11
79.92
80.73
81.55
82.36
83.18
84.00
84.82
85.65
86.47
87.30
88.13
88.96
89.79
90.62
10.8
10.9
11.0
11.1
11.2
11.3
11.4
11.5
11.6
11.7
11.8
11.9
12.0
12.1
12.2
12.3
12.4
12.5
12.6
12.7
12.8
12.9
13.0
13.1
13.2
13.3
13.4
13.5
13.6
13.7
13.8
13.9
14.0
14.1
14.2
14.3
14.4
14.5
14.6
14.7
14.8
0.108
0.109
0.110
0.111
0.112
0.113
0.114
0.115
0.116
0.117
0.118
0.119
0.120
0.121
0.122
0.123
0.124
0.125
0.126
0.127
0.128
0.129
0.130
0.131
0.132
0.133
0.134
0.135
0.136
0.137
0.138
0.139
0.140
0.141
0.142
0.143
0.144
0.145
0.146
0.147
0.148
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FLEXCELL® INTERNATIONAL CORPORATION
APPENDIX 2: STAGEFLEXER® 28 MM LOADING STATION™ CONVERSION CHART
Press (-kPa)
% Elong
Strain
Press (-kPa)
% Elong
Strain
Press (-kPa)
% Elong
Strain
0.00
0.70
1.39
2.08
2.76
3.44
4.11
4.78
5.44
6.10
6.76
7.41
8.05
8.69
9.33
9.97
10.60
11.22
11.85
12.47
13.08
13.70
14.31
14.92
15.52
16.12
16.72
17.32
17.91
18.51
19.10
19.68
20.27
20.85
21.43
22.01
22.59
23.17
23.75
24.32
24.89
25.47
26.04
26.61
27.18
27.75
28.32
28.88
29.45
30.02
30.59
31.16
31.72
32.29
0.0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0.9
1.0
1.1
1.2
1.3
1.4
1.5
1.6
1.7
1.8
1.9
2.0
2.1
2.2
2.3
2.4
2.5
2.6
2.7
2.8
2.9
3.0
3.1
3.2
3.3
3.4
3.5
3.6
3.7
3.8
3.9
4.0
4.1
4.2
4.3
4.4
4.5
4.6
4.7
4.8
4.9
5.0
5.1
5.2
5.3
0.000
0.001
0.002
0.003
0.004
0.005
0.006
0.007
0.008
0.009
0.010
0.011
0.012
0.013
0.014
0.015
0.016
0.017
0.018
0.019
0.020
0.021
0.022
0.023
0.024
0.025
0.026
0.027
0.028
0.029
0.030
0.031
0.032
0.033
0.034
0.035
0.036
0.037
0.038
0.039
0.040
0.041
0.042
0.043
0.044
0.045
0.046
0.047
0.048
0.049
0.050
0.051
0.052
0.053
32.86
33.43
34.00
34.57
35.14
35.71
36.29
36.86
37.44
38.01
38.59
39.17
39.75
40.33
40.92
41.51
42.10
42.69
43.28
43.88
44.47
45.07
45.68
46.28
46.89
47.51
48.12
48.74
49.36
49.99
50.62
51.25
51.89
52.53
53.17
53.82
54.48
55.13
55.79
56.46
57.13
57.81
58.49
59.17
59.86
60.56
61.26
61.97
62.68
63.40
64.12
64.85
65.59
66.33
5.4
5.5
5.6
5.7
5.8
5.9
6.0
6.1
6.2
6.3
6.4
6.5
6.6
6.7
6.8
6.9
7.0
7.1
7.2
7.3
7.4
7.5
7.6
7.7
7.8
7.9
8.0
8.1
8.2
8.3
8.4
8.5
8.6
8.7
8.8
8.9
9.0
9.1
9.2
9.3
9.4
9.5
9.6
9.7
9.8
9.9
10.0
10.1
10.2
10.3
10.4
10.5
10.6
10.7
0.054
0.055
0.056
0.057
0.058
0.059
0.060
0.061
0.062
0.063
0.064
0.065
0.066
0.067
0.068
0.069
0.070
0.071
0.072
0.073
0.074
0.075
0.076
0.077
0.078
0.079
0.080
0.081
0.082
0.083
0.084
0.085
0.086
0.087
0.088
0.089
0.090
0.091
0.092
0.093
0.094
0.095
0.096
0.097
0.098
0.099
0.100
0.101
0.102
0.103
0.104
0.105
0.106
0.107
67.08
67.83
68.59
69.36
70.13
70.91
71.70
72.49
73.29
74.10
74.91
75.73
76.56
77.40
78.25
79.10
79.96
80.83
81.71
82.59
83.48
84.39
85.30
86.21
87.14
88.08
89.02
89.98
90.94
10.8
10.9
11.0
11.1
11.2
11.3
11.4
11.5
11.6
11.7
11.8
11.9
12.0
12.1
12.2
12.3
12.4
12.5
12.6
12.7
12.8
12.9
13.0
13.1
13.2
13.3
13.4
13.5
13.6
0.108
0.109
0.110
0.111
0.112
0.113
0.114
0.115
0.116
0.117
0.118
0.119
0.120
0.121
0.122
0.123
0.124
0.125
0.126
0.127
0.128
0.129
0.130
0.131
0.132
0.133
0.134
0.135
0.136
6
FLEXCELL® INTERNATIONAL CORPORATION
APPENDIX 3: STAGEFLEXER® 31 MM LOADING STATION™ CONVERSION CHART
Press (-kPa)
% Elong
Strain
Press (-kPa)
% Elong
Strain
0.00
0.37
0.79
1.25
1.76
2.31
2.91
3.54
4.22
4.94
5.70
6.49
7.32
8.18
9.08
10.01
10.98
11.97
13.00
14.05
15.13
16.23
17.36
18.51
19.69
20.89
22.11
23.34
24.60
25.87
27.16
28.46
29.78
31.11
32.45
33.80
35.16
36.53
37.90
39.28
40.67
42.05
43.44
44.84
46.23
47.62
49.00
50.39
51.77
53.14
54.51
55.87
57.22
0.0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0.9
1.0
1.1
1.2
1.3
1.4
1.5
1.6
1.7
1.8
1.9
2.0
2.1
2.2
2.3
2.4
2.5
2.6
2.7
2.8
2.9
3.0
3.1
3.2
3.3
3.4
3.5
3.6
3.7
3.8
3.9
4.0
4.1
4.2
4.3
4.4
4.5
4.6
4.7
4.8
4.9
5.0
5.1
5.2
0.000
0.001
0.002
0.003
0.004
0.005
0.006
0.007
0.008
0.009
0.010
0.011
0.012
0.013
0.014
0.015
0.016
0.017
0.018
0.019
0.020
0.021
0.022
0.023
0.024
0.025
0.026
0.027
0.028
0.029
0.030
0.031
0.032
0.033
0.034
0.035
0.036
0.037
0.038
0.039
0.040
0.041
0.042
0.043
0.044
0.045
0.046
0.047
0.048
0.049
0.050
0.051
0.052
58.56
59.89
61.21
62.51
63.80
65.07
66.33
67.56
68.78
69.98
71.15
72.31
73.44
74.54
75.62
76.67
77.69
78.69
79.65
80.58
81.48
82.35
83.18
83.97
84.72
85.44
86.12
86.76
87.35
87.90
88.41
88.87
89.29
89.66
89.98
90.25
5.3
5.4
5.5
5.6
5.7
5.8
5.9
6.0
6.1
6.2
6.3
6.4
6.5
6.6
6.7
6.8
6.9
7.0
7.1
7.2
7.3
7.4
7.5
7.6
7.7
7.8
7.9
8.0
8.1
8.2
8.3
8.4
8.5
8.6
8.7
8.8
0.053
0.054
0.055
0.056
0.057
0.058
0.059
0.060
0.061
0.062
0.063
0.064
0.065
0.066
0.067
0.068
0.069
0.070
0.071
0.072
0.073
0.074
0.075
0.076
0.077
0.078
0.079
0.080
0.081
0.082
0.083
0.084
0.085
0.086
0.087
0.088
7