Caliper (Zymark) Pin Tools

     
   
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Defining a Pin tool
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Pin Tool Weights
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Caliper (Zymark) Sciclone Adaptor To Turn A
Sciclone ALH™  Liquid Handling System Into A Pin Tool

 


As the trend towards assay miniaturization continues with increasing demands to make nanoliter transfers, here is a simple way to convert a Sciclone ALH Liquid Handling System into a 96, 384 or 1,536 pin tool work station that will deliver from 2 nl to 5 ul.  You can easily switch back and forth on the fly between the Pipeting System and the pin tool.   The pin tool is hot swappable with both fixed cannula and disposable tip arrays.  Just park the Pin Tool on a V&P Docking Station (VP 550) and use the robot to dock with it and pick it up.

           
 

Caliper (Zymark) and V&P Scientific are collaborating to combine V&P's pin tool technology  with the high throughput automation of Calipers' (Zymark) Zephyr workstation. 
 

Pricing and system assistance is available from Caliper (Zymark) at
1-(508) 435-3439 or visit http://www.caliperls.com/products/zephyr-compact-liquid-handling-workstation.htm

 

PDF file on the Zephyr.

   
 


Caliper (Zymark) and V&P Scientific are collaborating to combine V&P's pin tool technology  with the high throughput automation of Calipers' (Zymark) Sciclone ALH workstation. 
 

Pricing and system assistance is available from Caliper (Zymark) at
1-(508) 435-3439 or visit http://caliperls.com/products/sciclonealh3000.html

 

PDF file on the Sciclone ALH 3000.

 

Click here to view a video demonstrating a V&P Pin Tool being used on a Sciclone ALH 3000.

 
 
 


Save Money

     
 


Pin tools will save you money.  Not only will you use less of your valuable lead compounds,  you will also pay for the pin tool in your second screening operation by eliminating the cost of disposable pipet tips.  At $.12/tip, 200 cases of 384 tips are ~$9,000.00

 
 
 
 


Cleaning The Pins

     
 


Procedures for transferring liquids, robot speed, wash solutions, blotting protocols and pin cleaning can found at this hyperlink.  Cleaning the pins between mother plates is easy and effective.  In an experiment to determine the amount of carry over after 100 transfers of concentrated FITC (mother to daughter plate) we were only able to detect  0.000,001,8% of the residual FITC on the pin using only two static 100 ml wash baths.  To put this into perspective, it is equivalent to removing all but 18 molecules out of a possible 1,000,000,000 molecules. This experiment was repeated using DMSO as the first wash solution and virtually identical results were obtained, indicating the thoroughness of the wash.

     
 


For comprehensive data on nanoliter dispensing with pin tools see the following pages.

 
 

 

Storage Stations for Pin Tools offer a safe place to store pin tools and keep them clean.

 


  Factors Affecting Pin Delivery Volumes

     
 


The list on the right summarizes the factors that contribute to the volume delivered.  With each application these factors can be controlled and standardized so that the delivery volumes are very reproducible.  With most applications the CV's are less than 5%.
 

 1. Pin Diameter
 2. Surface tension of the liquid being transferred
 3. Surface tension of the pin
 4. Speed of removal of pin from source liquid
 5. Speed of pin striking recipient dry plate
 6. Depth to which the pin is submerged in source plate
 7. Depth to which the pin is submerged in recipient plate
 8. Volume of slot in pin
 9. Surface tension of the dry plate and dwell time
     
 


V&P pins transfer liquid by carrying the liquid on the sides, tips and in slots we cut into the tips. The amount delivered is determined by the diameter and surface tension of the pin, the surface tension of the liquid, how far the pin is submerged into the liquid and the speed with which the pin is removed from the liquid.  When all of these variables are fixed, the accuracy of the liquid transfer is very consistent, with CV averages better than 5%.

 

Increasing the speed of withdrawal from the source plate by 7- fold will increase the volume delivered by as much as 3 fold in a linear relationship.  This phenomenon can be exploited to expand the range of delivery volume for a single pin. The table and the chart below illustrates the effects of increasing the speed 7- fold.  The speed range is from 0.78 cm/sec to 5.7 cm/sec.  Use this chart to select the pin that will deliver in your desired range.

     


  Select The Volume You Need To Transfer For Your Application

 
 


DMSO Liquid To Liquid Transfers With Uncoated Pins

     
   
Pin Diameter Shape Delivery Volume Range
FP9 0.229 mm Solid 5 - 15 nl
FP8 0.350 mm Solid 12 – 28 nl
FP1 0.457 mm Solid 19 –  68 nl
FP1S6 0.457 mm 6 nl Slot 26 – 74 nl
FP1S10 0.457 mm 10 nl slot 31 – 79 nl
FP1S20 0.457 mm 20 nl slot 43 – 89 nl
FP1S30 0.457 mm 30 nl slot 53 – 99 nl
FP1S40 0.457 mm 40 nl slot 64 – 109 nl
FP1S50 0.457 mm 50 nl slot 74 – 117 nl
FP3 0.787 mm Solid 84 – 240 nl
FP4 0.910 mm Solid 112 – 207 nl
FP 1.580 mm Solid Pointed 157 – 595 nl
FP3S100 0.787 mm 100 nl slot 178 – 326 nl
FP2S200 0.787 mm 200 nl slot 278 – 424 nl
FP6 1.580 mm Solid Flat 363 – 884 nl
FP3S500 0.787 mm 500 nl slot 527 – 656 nl
FPS.5 1.580 mm 0.5 ul slot 443 – 906 nl
FP6S.5 1.580 mm 0.5 ul slot 726 – 1,253 nl
FPS 1.580 mm 1 ul slot 938 – 1,424 nl
FP6S 1.580 mm 1 ul slot 1,239 – 1,786 nl
FPS2 1.580 mm 2 ul slot 1,817 – 2,248 nl
FP6S2 1.580 mm 2 ul slot 2,102 – 2,576 nl
FPS5 1.580 mm 5 ul slot 4,475 – 4,828 nl
FP6S5 1.580 mm 5 ul slot 4,475 – 4,828 nl
 
   

*Delivery volume range is determined by speed of withdrawal from source liquid.
slow speed  = 0.78 cm/sec = low volume delivery range
fast speed  =  5.70 cm/sec = high volume delivery range
Follow this link to assay methods

*Speed of withdrawal determines volume transferred. The faster the withdrawal speed the larger the volume transferred.  Withdrawal speed range is from 1.5 mm/sec to 30mm/sec.

 

*The volume of liquid in the source well also makes a significant difference in the volume transferred and there is a difference between sold and slotted pins.  The volume used for the 96 well data was 200 ul and the volume used for the 384 well data was 75 ul.  If you use more or less volume in your wells it will affect the volume transferred.  See this link for more information.

 

The links below are to delivery volume range tables for uncoated and hydrophobic coated delivering either DMSO or Aqueous solutions in liquid to liquid transfers.

 

DMSO TRANSFERS WITH HYDROPHOBIC COATED PINS

 

DMSO TRANSFERS WITH UNCOATED (STAINLESS STEEL) PINS

 

AQUEOUS TRANSFERS WITH HYDROPHOBIC COATED PINS

 

AQUEOUS TRANSFERS WITH UNCOATED (STAINLESS STEEL) PINS

   


  A Guide For Selecting The Right Pin For Your Application

     
 


 

1.  What volume do you need to  transfer?   DMSO? Aqueous? See these hyperlinked tables to determine your options.  Also consider custom slot pins.

 

2. Will the source plate have wells with significantly different levels of liquid?  (Cherry picked source plate or edge drying effect?).  If yes and if the absolute volume transferred is critical, then select the largest slot pin that is in your transfer range.  This will minimize the effect of liquid height on the volume of liquid carried on the sides of the pin.  Also consider custom slot pins.

 

3. Does the material transferred bind non-specifically to stainless steel?  If yes then select the Hydrophobic/lipophobic coated pins.  If no, select the uncoated pins.

 

4.  What is the Z clearance on the robot deck from the highest impediment (top of the source plate/recipient plate/wash reservoirs, etc.) to the top of the robot mounting plate?  If it is greater than 77 mm you can use any of pins.  If it is less than 77 mm, you can only use the shorter FP(#)C series pins (12 mm exposed pin length), the FP(#N) series pins (17 mm exposed pin length) and the E-clip series pins (23 mm exposed pin length pins.  If the Z clearance is less than 60 mm, we recommend you contact us.  There are a few tricks we can do to shave off 5 to 10 mm in height. 

 

Links to the specifications of all the robot mounting plates/pin tool/pin combinations can be found on the height ("Z") restriction page

 

5.  Is the source plate a deep well plate?  If so you may need to use a 30 mm long exposed length pin ("T" pin) to reach down to the lower levels of the well. 

 

6.  Should you choose the solid pin versus the slot pin? Although the there appears to be a slight advantage for slot pin, CV's between the two pins are very good.  Both are easy to clean between specimens.  Biggest factor is cost.  If you don't need to deal with varying liquid heights and you can obtain the volume necessary with a solid pin, choose the economical solid pin.

 

7.    If you are still uncertain about which pin to select for your application, you can perform a simple "Proof of Principle" test with several different pins using our inexpensive VP 450FP3 Replicator Strip coupled via a VP 452MP to one of our robot mounting plates.   An even simpler solution is to use a work station with a 1, 4 or 8 probe dispense head with the new V&P Mono Pin Tool to test the various pins.

 


  Liquid To Dry Plate Transfers For Multiplexing Assays

     
   

Many of our customers want to make liquid to dry plate transfers to make mini-microarrays in a single well for multiplexing assays.  By their very nature these transfers only leave a small volume remaining on the plate.  The amount remaining depends on the surface tension of the plate, the transfer liquid and the pin (diameter and shape).  We have found that reasonable results can be obtained if you use an aqueous spotting buffer and a polypropylene surface (CV's less than 9%).  DMSO spotting buffers leave much less liquid on the dry plate and when used with polystyrene plates the CV's are significantly higher. 

 

The plate to the right was spotted using a 50 nl slot pin (FP1CS50) transferring oligo DNA in an aqueous spotting buffer to a specially treated polypropylene plate.  Our customer in this case wanted to place 42 individual spots in the bottom of each well of a 96 well microplate.  Spot diameter = 0.55 mm. Volume transferred to the spot = 5.85 nl

 

The photo on the right is the result of a four fold serial dilution hybridization assay of the Oligo spots with a labeled DNA that can be detected optically and quantitated.

 

Follow this link for more information on liquid to dry transfers and Multiplexing.

 

 


  Defining A Pin Tool

     
 


Originally we sold our pin tools with unique part numbers.  As we have added more pins to our selection and more robot systems to mount the pin tools to, the number of unique part numbers grew exponentially to an un-manageable size.  Our solution has been to specify a pin tool by the 3 component parts that compromise the Pin Tool.  The Pins, the Floating Fixture and the Robot Mounting Plate.  If you have previously ordered a pin tool by unique part number, we will be able to provide the same pin tool to you.

     


  Ordering Your Sciclone Pin Tool As Simple As 1, 2, 3

     
 


A pin tool consists of three separate parts.

 

1.  The pins, (#FP___)
2.  The Floating Frame Fixture (96, 384 or 1536) (#AFIX___FP_)
3.  The mounting plate that attaches the Floating Frame Fixture to the Zymark Sciclone (#BMPZYMARK)

 

For example the part numbers for pin tool in the photo to the right would be FP3, AFIX384FP3 and BMPZYMARK.

 

To order, just tell us those 3 part numbers and we will assemble your Pin Tool.

 
 


  1. Choose the pin to match your application, pin length, delivery volume requirements
  and/or the non-specific binding nature of the sample.

 
 


Solid Pins

     
 


Solid pins are the most economical and control volume transferred by pin diameter.

 

Solid pins are available in 5 different exposed pin lengths; 12, 17, 22, 23 and 30 mm.  The 12 mm ("C") pins are used with robots that have "Z" axis limitations and the 30 mm ("T") pins are used when transferring from a deep well source plate.  The E-clip pins have exposed lengths of 22 and 23 mm.  They are used to transfer larger volumes of liquids because of their larger diameter

     
 
Slot Pins
     
 


Slot pins have several advantages over solid pins:

  1. Slot pins carry a larger volume than solid pins in liquid to liquid transfers.

  2. Slot pins are not as sensitive to the depth of the mother liquid as solid pens.

  3. Slot pins deliver a larger volume than solid pins in liquid to dry plate transfers.

  4. Slot pins deliver a larger volume than solid pins in liquid to membrane transfers.

  5. Slot volume is customizable.

  6. Slot pins can deliver to dry plates by pre-wetting the plate.

     
 


Hydrophobic Drophobic/Liphobic Coated Pins

     
 


Hydrophobic/lipophobic coated pins are very useful in handling solutions with proteins or lipids that may stick to naked stainless steel.  Both solid and slot pins can be coated.  Click here for more details on the nature of our hydrophobic/ lipophobic coating and the coating process.

     
 


The tables below contain hyperlinks to the various types/lengths/diameters/coatings of the pins we offer.

     
 


Tube Style Pins

     
   

Uncoated

12 mm/33 mm length - solid pins 17 mm/38 mm length solid pins 17 mm/50 mm length solid pins 30 mm/50 mm length solid pins
12 mm/33 mm length slot pins 17 mm/38 mm length slot pins 17 mm/50 mm length slot pins 30 mm /50 mm length slot pins
 
   

Hydrophobic coated

12 mm/33 mm length solid pins 17 mm/38 mm length solid pins 17 mm/50 mm length solid pins 30 mm/50 mm length solid pins
12 mm/33 mm length slot pins 17 mm/38 mm length slot pins 17 mm/50 mm length slot pins 30 mm/50 mm length slot pins
 
 


E-Clip Style Pins
 

     
     
 

Please visit this hyperlink for Pin Tool Data And Charts on hydrophobic pins.

     


  2. Choose the Floating Fixture to match your application.  Standard are 96, 384 and 1,536
  formats.

 
 


These floating fixtures consist of an upper float plate that will all accommodate a 1.58 mm diameter tube, a lower float plate that will accommodate either a .229 mm (FP9), .457 mm (FP1) or .787 mm (FP3) diameter pin and 6 standoffs that keep the two plates separate.  The length of the standoffs is determined by the length of the pin used.  Your application will determine if you select the 96, 384 or 1,536 formats and your previously made pin selection will determine the diameter of holes in the lower float plate.  Click here for a complete selection of these floating fixtures..

 

If you have more specific needs call us for help at (800) 455-0644.

 
 
 


Combine the number and type of Pins with the Floating Frame Fixture selected and the Basic Mounting Plate for the Caliper (Zymark) Sciclone (BMPZYMARK) and you have a complete pin tool. 

 

Call Caliper (Zymark) for pricing information at 1-(508) 435-3439 and tell them the part numbers you need for the pin tool - Pin # (FP___)
Floating Frame Fixture # (AFIX__FP__ )
Zymark Sciclone Mounting Plate # (BMPZYMARK).

 
 
 


The Caliper (Zymark)/V&P pin tool above has 384 FP3 pins that are 0.787 mm in diameter and will deliver ~ 80 nl

 
 
 


The Caliper (Zymark)/V&P pin tool above has 1,536 FP3 pins that are 0.787 mm in diameter and will deliver ~ 80 nl

 
 


  Replacement Pin Link for the Tube style TekCel Pin Tools.

 
 
The Double Float Plate TekCel Assay TekBench Tube Style Pin Tools have several advantages over the E-Clip Style Pin Tools:
  1. The Tube Style Pin Tools are smaller in diameter and deliver very small volumes (down to 2 nl and as much as 500 nl depending on diameter and slot size).

  2. Smaller pin diameters also result in more precise delivery volumes (CV's ~4%).

  3. Because the pin is registered between 2 float plates over a greater height, the pin angle will be truer (closer to 90) when they touch the surface. 
  4. The smaller hole on the lower float plate allows us to make very tight tolerances on the fit between the pin and the hole.  This results in very precise placement of the pins on the surface so that very high density arrays can be made without overlapping the spots.

The E-Clip Pin Tools on the other hand are capable of delivering larger volumes (from 0.5 to 5 ul).

     


  Custom Floating Slot Pin Replicators

     
 


Custom Floating Slot Pin Replicators can also be made using the 0.457 mm and 0.787 mm diameter pins and placing them into the Floating Pin Replicators described above.  The slot volumes between these two pins can cover a range of volumes from 5 nl to 2000 nl by changing the width and depth of the slot.  Inquire by phone or e-mail for more information.

     


  Cleaning Pins On A Caliper (Zymark) Sciclone Mounted Pin Tool

 
 


The most important aspect of using pin tools is to start with clean pins and then keep them clean so they will consistently deliver the same volume.  We recommend our Pin Cleaning Solution (VP 110) to clean the pin of protein and other residue buildup before you start an assay.  To quickly and efficiently remove the previous transfer from the pin we have several different wash reservoirs, flowing reservoirs and fountains to clean  96, 384 and 1536 floating pin tools.  These wash stations also are adapted so the blotting station is part of the washing station thus reducing the cleaning cycle time and the number of robot stations required for the cleaning process.  We also have other independent blotting stations (VP 540D).


Integral to the cleaning process of all pins is the blotting step between baths.  Lint particles in the slots or on pin tips, from standard paper towels, will interfere with the loading and unloading of liquid.  We have solved this problem.  The VP 540DB blotting system is a tray with the standard microplate foot print that contains a super absorbent polypropylene pad covered by a Lint Free Blotting Paper.  We strongly recommend that you use this or our other lint free blotting systems.

     
 


Storage Stations for Pin Tools offer a safe place to store pin tools and keep them clean and out of harms way.

     


  Pin Tool Accessory Kit

     
 


We highly recommend that you order the Caliper (Zymark) Pin Tool Accessory kit at the time you order the pin tool.  This "essentials" accessory kit contains the VP 550 Docking Station for the Pin Tool on the robot deck giving you the capability for "Hot Swapping" the pin tool for other accessories, The VP 540DB Lint Free Blotting System is used to clean the pins efficiently between samples, two VP 540 Wash Reservoirs to wash the pins between source plates, the VP 110 Pin Cleaning Solution is used the clean the pins daily, the VP 425 Pin Cleaning Brush is used if a build up occurs and the VP 555 Safe Pin Tool Storage System used to keep it clean and the fragile pins out of harms way.   Although we have several other wash and blot accessories available these are the most cost effective for the majority of applications.  If you elect to use the fountain and flowing reservoirs we still strongly recommend the other items in this kit.  When purchased as the VP OK18 Kit you will receive a 5% discount over the sum of the individual items.

     
   

Zymark Sciclone Pin Tool Accessory Kit - VP OK18

Pin Tool Docking Station for "hot swapping" (1) VP 550
Lint Free Blotting System with pad (1) VP 540DB
Pin Wash Reservoirs (2) VP 540
Pin Cleaning Solution (1) VP 110
Pin Cleaning Brush (1) VP 425
Safe Pin Tool Storage Station (1) VP 555
     


  Solid Floating E-Clip Pins

     
 


E-Clip pins are ideally suited for delivering larger volumes of liquids from one plate to another or to a membrane.

 
 


  Solid E-Clip Pin used in Double Float Plate Replicators

     
   
Exposed pin length = 23 mm, overall pin length = 34 mm, pin diameter = 1.58 mm, pointed tip, solid pin hanging drop = ~100 nl FP
Exposed pin length = 22 mm, overall pin length = 33 mm, pin diameter = 1.58 mm, flat tip, solid pin hanging drop = ~400 nl FP6
 


 Slotted Floated E-Clip Pins

     
 


**NEW** Flat Tip Slotted E-Clip Pins

     
 


Our new FP6 slotted flat tip E-Clip pins are the pins of choice for transferring liquids to agar or other soft surfaces and melting/transferring frozen stocks while replicating genomic/microbial libraries.

 
 
   
Exposed pin length = 22 mm, overall pin length = 33 mm, pin diameter = 1.58 mm, flat tip, slot volume = 0.5 ul FP6S.5
Exposed pin length = 22 mm, overall pin length = 33 mm, pin diameter = 1.58 mm, flat tip, slot volume = 1.0 ul FP6S
Exposed pin length = 22 mm, overall pin length = 33 mm, pin diameter = 1.58 mm, flat tip, slot volume = 2.0 ul FP6S2
Exposed pin length = 23 mm, overall pin length = 33 mm, pin diameter = 1.58 mm, flat tip, slot volume = 5.0 ul FP6S5
 
 


Pointed Tip Slotted E-Clip Pins

     
 


Our pointed tip FP series slotted E-Clip pins are used to remove samples from the tips of conical wells of source plates.  Because of the sharp point they are not recommended for spotting on agar or other soft surfaces.

 

 
 
 


Pointed E-Clip Slot Pins used in Double Float Plate Replicators


VP 408FS.5, VP 408FS, VP 408FS2, VP 408FS5, VP 384FS.5, VP 384FS,
VP 384FS2, VP 384FS5 

     
   
Exposed pin length = 23 mm, overall pin length = 34 mm, pin diameter = 1.58 mm, slot volume = 0.5 ul FPS.5
Exposed pin length = 23 mm, overall pin length = 34 mm, pin diameter = 1.58 mm, slot volume = 1 ul  FPS1
Exposed pin length = 23 mm, overall pin length = 34 mm, pin diameter = 1.58 mm, slot volume = 2 ul  FPS2
Exposed pin length = 23 mm, overall pin length = 34 mm, pin diameter = 1.58 mm, slot volume = 5 ul  FPS5
 
 


**The actual volumes delivered depend upon many controllable factors that will vary from one application to another.  For example in liquid to liquid transfers the amount transferred will include the hanging drop on the bottom of the pin as well as the liquid on the sides of the pin.  If the transfer was to a membrane then only the hanging drop would be transferred and not the liquid on the sides of the pin.  If the transfer was to a dry plate then only about 33% the hanging drop would be transferred to the dry plate and the rest would be on the pin.  The division of the hanging drop would be determined by the surface tension of the pin and the surface tension of the dry plate

     
 


:-)

 
         

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