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Many of our customers have used our pin tools to "wound" cell
culture monolayers and then study the effects of different
treatments on wound healing. Initially our VP 408FH Replicator, VP
384FH Replicator or Robotic Pin Tools converted to manual use with
the BMPM-PAD-1 plus a special wounding Library Copier (VP 381NW, VP
381NW4.5 or VP 381NW5) were used to make the wounds. For most
applications, these wounding tools give good results. However, some
customers who were using computerized image analysis programs needed
the wounds to be more precisely localized. For these customers, we
have developed specialized pin tools that have very tight hole
tolerances in the floating fixture so the wounds are precisely and
consistently located in the same position in each well. These
wounding series pin tools (our “W” and “WP” pin tools) are
illustrated on this page.
A recent addition to our line of wounding pin tools are pins with a 0.05 mm
layer of Parylene deposited using a vapor deposition process to produce a soft
lining on the pin tips. Each pin now acts as “eraser” to remove the cell
monolayer without scratching the plastic well surface under the monolayer.
Another new innovation and problem solved only from V&P Scientific.
The wounding pin tools that incorporate Parylene-coated pins can also be used in
our standard floating fixtures because the pins are larger in diameter and fit
snugly in the standard fixture holes. In contrast, the uncoated wounding pins
need to have the smaller diameter holes found in the wounding floating fixtures
in order to achieve the snug fit.
Another factor that can lead to wound variation is a loose fit between the
microplate and the registration device (e.g., our VP 381NW Library Copier).
This is due to variation in the molds used to make the microplates. One way of
dealing with the problem is to use our adjustable Library Copier (VP 381NWGV4.5).
A less elegant but practical solution is to apply layers of tape to the inside
of our standard wounding Library Copiers until you have a snug fit with the
microplate.
Click here for a list of published articles that cite the products on this page.
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The photo to the right shows the
wounding dimensions of VP 381NW,
VP 381NW4.5 and VP 381NW5 with FP,FP6, FP6-WP, FP4-WP and
FP1-WP pins.
This is how the Small Molecule Screening Facility at Keck-UWCCC does their
quantitative wound healing assay using VP 408FH.
http://www.hts.wisc.edu/Resources.htm#wh |
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Part # |
Wound
width in mm** |
Requires Standard or Wounding Floating Fixture |
Coated with Parylene |
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FP-W |
0.37 mm to 0.51 mm |
Wounding Fixture |
No |
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FP1-W |
0.457 mm to 0.458
mm |
Wounding Fixture |
No |
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FP4-W |
0.914 mm to 0.915
mm |
Wounding Fixture |
No |
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FP6-W |
1.58 mm to 1.59 mm |
Wounding Fixture |
No |
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FP-WP |
0.47 mm to 0.61 mm |
Standard Fixture |
Yes |
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FP1-WP |
0.557 mm to 0.558 mm |
Standard Fixture |
Yes |
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FP4-WP |
1.014 mm to 1.015 mm |
Standard Fixture |
Yes |
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FP6-WP |
1.68 mm to 1.69 mm |
Standard Fixture |
Yes |
** The wound width will also be determined
by the type of cell monolayer and how many passes are made.
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The photos to the right shows a 96
Wounding Replicator with FP1-WP Pins.
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E-Clip Style Floating, Solid Pin - 1.58mm Chamfered to 0.3mm Diameter Flat
Tip, 34mm Long, 23mm Exposed Pin Length, Delivers ~100nl |
FP |
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Parylene coated wounding pin 1.68 mm Diameter Pin with a 0.481 mm to 0.608
mm diameter point 35 mm Long with an E-Clip Groove |
FP-WP |
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E-Clip Style Floating, Flat Tip Solid Pin - 1.58mm Diameter, 33.3mm Long,
22.5mm Exposed Pin Length |
FP6 |
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Parylene coated wounding pin 1.68 mm Diameter Pin 35 mm Long with an E-Clip
Groove, price each |
FP6-WP |
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Wounding Pin 0.457 mm Diameter Pin Protruding 1 mm Out a 38 mm Long Cannula,
1.58 mm in Diameter with an E-Clip Groove in the Cannula |
FP1-W |
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Parylene Coated Wounding Pin with 0.557 mm Diameter Pin Protruding 1 mm Out
a 38. mm Long Cannula, 1.68 mm in Diameter with an E-Clip Groove in the
Cannula |
FP1-WP |
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Wounding Pin 0.914 mm Diameter Pin Protruding 1 mm Out a 38 mm Cannula with
an E-Clip Groove in the Cannula |
FP4-W |
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Parylene Coated Wounding Pin with 1.014 mm Diameter Pin Protruding 1 mm Out
a 38 mm Long Cannula, 1.68 mm in Diameter with an E-Clip Groove in the
Cannula |
FP4-WP |
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Foam silicone rubber gasket used to hold the pins evenly on
the surface of the mono-layer.
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Other useful and highly recommended accessories used with
wounding pin
tools are
the VP 426 Pin Cleaning Pad and VP 540S
Reservoir for holding the Pin Cleaning
Pad while
cleaning the pins between woundings.
VP 426 - Pin Cleaning
Pad, 4 mm nylon
bristles.
VP 540S
- Polypropylene reservoir holding a VP 426 Pin Cleaning
Pad for pin cleaning. SBS feature on the bottom for
deck placement. |
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The AFIX24FP6W was designed to be used in tissue
culture "wounding" experiments where a monolayer of tissue
culture cells was grown in a 24 well microplate and a V&P
floating pin replicator was used to create a precise and
reproducible wounding pattern in all the mono-layers. The
scientists could then apply different treatments to each
well and gauge how effective they were.
The pins on this replicator are on an 19.3 mm center to center
spacing. Each 24 well microplate manufacture has a
slightly different center to center spacing, please check
before ordering.
In
addition to the AFIX24FP6W floating frame you need to order
the BGPK-E Kit, either 24 FP or 24 FP6 pins and the BMPM-PAD-1
to obtain a complete replicator. For large (1.58 mm
wide) wounds choose the FP6 pins. For smaller (0.38 mm
wide) wounds choose the FP pins.
The
AFIX24FP6W is made with slightly smaller (1.61 mm) diameter holes in
the float plate than the AFFIX24FP6 so the pins will not
"wobble" when wounding the mono-layer.
The
following 24 well microplates have 19.3 mm center to center
spacing and are compatible with our wounding replicator:
BD
Biosciences (Falcon) Cat. No.s 353047, 353226,
353935,353847, 351147, 358115, 353504
Corning
life Sciences Cat. No.s 3337, 3524, 3526, 3527, 3473
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The AFIX48FP6W was designed to be used in tissue
culture "wounding" experiments where a monolayer of tissue
culture cells was grown in a 48 well microplate and a V&P
floating pin replicator was used to create a precise and
reproducible wounding pattern in all the mono-layers. The
scientists could then apply different treatments to each
well and gauge how effective they were.
The pins on this replicator are on an 13.5 mm center to
center spacing (Nunc plates). Each 48 well microplate
manufacture has a slightly different center to center
spacing, please check before ordering.
In addition to
the AFIX48FP6W floating frame you need to order the BGPK-E Kit,
either 48 FP or 48 FP6 pins and the BMPM-PAD to obtain a complete
replicator. For large (1.58 mm wide) wounds choose the FP6
pins. For smaller (0.38 mm wide) wounds choose the FP pins.
The AFIX48FP6W is made with slightly smaller
(1.61 mm) diameter holes in the float plate than the AFFIX48FP6 so the pins
will not "wobble" when wounding the mono-layer. |
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The AFIX96FP6W was designed to be used in tissue
culture "wounding" experiments where a monolayer of tissue
culture cells was grown in a 96 well microplate and a V&P
floating pin replicator was used to create a precise and
reproducible wounding pattern in all the mono-layers. The
scientists could then apply different treatments to each
well and gauge how effective they were.
The pins on this replicator are on 9 mm center to center
spacing.
In addition to
the AFIX48FP6W floating frame you need to order the BGPK-E Kit,
either 96 FP or 96 FP6 pins and the BMPM-PAD-1 to obtain a complete
replicator. For large (1.58 mm wide) wounds choose the FP6
pins. For smaller (0.38 mm wide) wounds choose the FP pins.
The AFIX96FP6W is made
with slightly smaller (1.61 mm) diameter holes in the float plate than the
AFFIX96FP6 so the pins will not "wobble" when wounding the
mono-layer. |
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The AFIX384FP6W was designed to be used in tissue
culture "wounding" experiments where a monolayer of tissue
culture cells was grown in a 384 well microplate and a V&P
floating pin replicator was used to create a precise and
reproducible wounding pattern in all the mono-layers. The
scientists could then apply different treatments to each
well and gauge how effective they were.
The pins on this replicator are on 4.5 mm center to
center spacing.
In addition to
the AFIX48FP6W floating frame you need to order the BGPK-E Kit,
either 384 FP or 384 FP6 pins and the BMPM-PAD-1 to obtain a complete
replicator. For large (1.58 mm wide) wounds choose the FP6
pins. For smaller (0.38 mm wide) wounds choose the FP pins.
The AFIX384FP6W is made with slightly
smaller (1.61 mm) diameter holes in the float plate than the AFFIX384FP6 so
the pins will not "wobble" when wounding the mono-layer.
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The VP 381NW LIBRARY COPIER was
designed to be used in tissue culture "wounding" experiments where a
monolayer of tissue culture cells was grown in a 96 or 384 well microplate and a
V&P floating pin replicator was used to create a precise and
reproducible wounding pattern in all the mono-layers. We modified a
VP 381N into a VP 381NW by making it with 2 oval alignment holes (1.58 mm
by 2.58 mm) so the 2 guide pins
could be moved in a race track movement and the replicator pins would
damage the monolayer in a precise and reproducible manner. . The
scientists could then apply different treatments to each well and gauge
how effective they were. A variety of different pin
diameters on our Replicators allows for different patterns of damage.
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The photo to the right illustrates an oval alignment hole.
The first scientist to use wounding
monolayers with one of our floating pin tools was Justin C. Yarrow
at Harvard. In
Dr. Yarrow's experiments he used one of our 384 pin tools and did the wounding
"free hand". The VP 381NW is an improvement over the free hand
technique.
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The VP 381NW4.5 is the same as the VP
381NW except for having longer ovals (4.5 mm) which allows
for longer wounding pattern if desired.
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The VP 381NWGV4.5 was designed with an adjustable base so that it can be
registered precisely and securely to any microplate. Thus wounds that are made
using it are precise in length and width.
Because microplate dimensions vary from manufacture to manufacture and between
batches we have developed an adjustable registration frame to hold the
microplate securely so it can not move during the wounding process. |
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The photo on the left illustrates how the base can be precisely
adjusted to accommodate plates on slightly different dimensions. A
less elegant but practical solution is to apply layers of tape to
the inside of a VP 381NW4.5 until you have a snug fit to the
microplate.
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The VP 381NW5 is the same as the VP
381NW and VP 381NW4.5 except for having longer ovals (5 mm) which allows
for longer wounding pattern if desired.
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The VP 381NW5-V is the same as the VP
381NW5 except for having ovals (5 mm) in a vertical
orientation.
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The VP 381NWYR1.7 was designed to make a 1.7 mm long wound in
a tissue culture monolayer grown in 384 well microplates using a 96
pin wounding pin tool. The VP 381NWYR1.7 is composed of 2
registration frames (right and left handed versions) that are used
in tandem to make 4 wounding processes with the 96 pin tool. The
sides of the registration frame register to the sides of the pin and
keep it straight while making the wounds. This method produces
straight reproducible wounds.
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One of the advantages of this system is that you can use a less
expensive 96 pin wounding pin tool rather than a more expensive 384
pin wounding pin tool to accomplish the same wounding pattern.
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This registration system will work with any of our 96 pin wounding pin tools.
This system also contains alignment holes in the registration frame to help
register the pin tool to the microplate wells. |
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Another group wanted to see if
the number of cells in a monolayer would affect the wound healing so we
made another Library copier that would fit over 24 well microplates.
The 24 well microplates are taller than 96 and 384 well plates so they
required a custom Library Copier to accommodate them, the VP 381W-24.
The oval pattern dimensions are the same (1.58 mm by 2.58 mm) but in a
vertical orientation.
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