Cell Culture Wounding Applications

     
 


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.

     
 


Wounding Dimensions in Microplates

     
 


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

 
 

 


Wounding Pins

     
 
 
 
 
 


Pins Used in Wounding Experiments

 
   
Part #

Wound width in mm**

Requires Standard or Wounding Floating Fixture

Coated with Parylene

FP-W

0.37 mm to 0.51 mm

Wounding Fixture

No
FP1-W 0.457 mm to 0.458 mm Wounding Fixture No
FP4-W 0.914 mm to 0.915 mm Wounding Fixture No
FP6-W 1.58 mm to 1.59 mm Wounding Fixture No
FP-WP 0.47 mm to 0.61 mm Standard Fixture Yes
FP1-WP 0.557 mm to 0.558 mm Standard Fixture Yes
FP4-WP 1.014 mm to 1.015 mm Standard Fixture Yes
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.
 

 
 


The photos to the right shows a 96 Wounding Replicator with FP1-WP Pins.

 
 
     
 
   
E-Clip Style Floating, Solid Pin - 1.58mm Chamfered to 0.3mm Diameter Flat Tip, 34mm Long, 23mm Exposed Pin Length, Delivers ~100nl FP
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
E-Clip Style Floating, Flat Tip Solid Pin - 1.58mm Diameter, 33.3mm Long, 22.5mm Exposed Pin Length FP6
Parylene coated wounding pin 1.68 mm Diameter Pin 35 mm Long with an E-Clip Groove, price each FP6-WP
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
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
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
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
 
     


  BMPM-PAD-1

     
 
Foam silicone rubber gasket used to hold the pins evenly on the surface of the mono-layer.
 
 


  Pin Cleaning Accessories

     
 


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.

 

 


  Wounding Replicators

     


     24 Well Format

     


AFIX24FP6W

     
 


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

 

 

 
 


     48 Well Format

     


AFIX48FP6W

     
 


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.

 
 


     96 Well Format

     


AFIX96FP6W

     
 


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.

 
 


     384 Well Format

     


AFIX384FP6W

     


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.

 
 
         
 

AFIX384FPWP

     
         
 

The AFIX384FPWP was designed to be used in tissue culture "wounding" experiments. V&P Scientific’s Wounding Pin Tools are used to create a precise and reproducible wounding pattern in every well of the microplate. The scientists could then apply different treatments to each well and gauge how effective they were.  The AFIX384FPWP registers the pins at a 4.5mm center-to-center spacing. 


In addition to the AFIX384FPWP floating frame you need to order the following to obtain a complete replicator:
BMPM-E-CLIP
FP-WP (Qty 384) or FP6-WP (Qty 384)
BMPM-PAD (Qty 1) and BMPM-PAD-1 (Qty 2)


FP-WP will produce a 0.38 mm wide wound
FP6-WP will produce a 1.68 mm wide wound

 

 

 


  Library Copiers

     
 


VP 381NW

     
 


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.

 
 
 


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.
 

 
 
 



VP 381NW4.5

     
 


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.
 

 

 
 


VP 381NWGV4.5

     
 


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.

 
 
 


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.

 
 
       
 

VP 381NWGH4.5

     
         
 

The VP 381NWGH4.5 was designed with an adjustable base so that it can be registered precisely and securely to any microplate. Wounds can be made in the horizontal direction in both 96 and 384 well plates. Wounds made using it are precise in length and width.

 

This library copier has two sets of slots which allows the user to use a partially filled 384 pin tool fixture containing only 96 or 192 pins to create wounds in all 384 wells.

 



 



VP 381NW5

     
 


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.
 

 

 
 



VP 381NW5-V

     
 


The VP 381NW5-V is the same as the VP 381NW5 except for having ovals (5 mm) in a vertical orientation.

 

 

 
 


VP 381NWYR1.7

     
 


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.

 
 
 


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.

 
 
 


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.

 
 
     
 
     
 


VP 381W-24

     
 


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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