Cybio Pin Tools

     
   
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Select Mounting
Cleaning Pins
Blotting Station
Technotes
Storage Stations
Fixture Frames
Pin Tool Data
Defining a Pin Tool


Cybio Adapter to turn a  CYBI™ Well  Pipetting System into a Pin Tool Work Station*

 

 

As the trend towards assay miniaturization continues with increasing demands to make nanoliter transfers, here is a simple way to convert either a 96 or a 384 CyBi Well Pipetting 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 between the Pipetting System and the pin tool by just exchanging the adaptor for the tip manifold.

 

Cybio and V&P Scientific, Inc. are again collaborating to place V&P Pin tools onto CyBio's CyBiR Well workstations.  Contact CyBio Inc at productinfo@cybio-ag.com or ((877)879-5624) for pricing and availability.  You can learn more about their complete line of robotic workstations at the Cybio web site  (http://www.cybio-ag.com/english/index.html).   Please contact V&P Technical Service at sales@vp-scientific.com  or (800) 455-0644  for technical questions about the pin tools.

           


  SAVE MONEY

     
 


If you have an obsolete 96 or 384 CybiR-Well pipeting station or one with damaged pistons, you can bring it back on line as a 96, 384 or 1536 pin tool work station.  Not only will you save a ton of money rehabilitating an obsolete or broken work station,  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.

 
 


  Pin Resetting Feature

     
 


In addition our CyBio adaptor also uses the pistons on the CyBio Pipettor to reset the pins in the rare event that they get stuck in the up position.  Simply programming the aspirate and dispense functions to preset values determines correct travel of the pistons.  The pistons of the CyBiR-Well Pipetting System pass right through the holes in the adaptor and press on the push pad.  The Piston Push Pad is connected to the Pin Push Pad and motion is transferred to the pins (see photo to the right).

 

We also use a similar fixture with different actuating mechanism on the PerkinElmer Pin tools and the Beckman FX to reset the pins.

 
 


  Robot Selection and pin length

     
 


Whether you have a 96 or a 384 CyBiR-Well or a PlateMate  Pipetting System we can attach any of our 96, 384 or even 1536 pin tool formats to either a 96 or a 384 CyBio Adaptor.  Because of the height limitation of the CyBi Well Pipetting System we use a shorter pin and a shorter tube to reduce the overall pin length to 33 mm.  The pin protrudes 12 mm below the lower float plate.  The pins used in the CyBio  Double Float Plate Replicators are all fixtured in a 1.58 mm diameter tube to give strength and stability to the small diameter pins.   The total height of CyBio Replicators from the top of the adaptor plate to the tip of the FPC series pins is  59 mm.  We recommend the FPC series pins for the standard CiBiR-Well.  For deep well CyBiR-Wells you can use the taller FP series pins.  The total height of the CyBiR-Replicators with FP series pins is 77 mm.  When you place your order we will need to know which adaptor to use (96 or 384) and the pin length.

 

Technical notes section for details on how to program the CyBiR-Well for pin tool applications.

     


  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.  This is equivalent to reading of 18 out of a possible 1,000,000,000 units 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 below 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.

     


  Selecting a Pin Tool as Simple as 1, 2, 3

   


 
 


1. Select the appropriate pin.  For the standard microplate CyBio and Plate Mate robots we recommend the shorter FPC_ series pins.  For the deep well  microplate versions we recommend the  longer FP_ series pins.

 

2. Select the Floating Fixture with the appropriate pattern (96, 384 or 1,536) and diameter of holes.

 

3. Select the robot system you wish to use the Pin Tool with.  We have made Mounting Plates to for both the 96 and 384 dispensing heads.  The BMP Mounting Plate in  the photo to right will accommodate the CyBio 96 dispensers.

 

   


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

 


  Tube Style Pins

     
 


     
 


Uncoated

 
   
Pin Type Shape Exposed Pin Length Total Pin Length
FP_CB Solid 12 mm 33 mm
FP_N Solid 17 mm 38 mm
FP_ Solid 17 mm 50 mm
FP_T Solid 30 mm 50 mm
       
FP_CS_ Slot 12 mm 33 mm
FP_NS_ Slot 17 mm 38 mm
FP_S_ Slot 17 mm 50 mm
FP_TS_ Slot 30 mm 50 mm
 
 


Hydrophobic coated

     
   
Pin Type Shape Exposed Pin Length Total Pin Length
FP_CH Solid 12 mm 33 mm
FP_NH Solid 17 mm 38 mm
FP_H Solid 17 mm 50 mm
FP_TH Solid 30 mm 50 mm
       
FP_CS_H Slot 12 mm 33 mm
FP_NS_H Slot 17 mm 38 mm
FP_S_H Slot 17 mm 50 mm
FP_TS_H Slot 30 mm 50 mm
 


  E-Clip Style Pins

     
 

 

 
 


  48 Pin Double Float Plate CyBio Replicators

     
 


The pin tool on the right has 48 pins that are centered in the replicator.

 
 


  96 Pin Double Float Plate CyBio Replicators

     
 


T
he pin tool to the right has 96 pins that are .457 mm in diameter and deliver

~ 20nl

 
 


  384 Pin Double Float Plate CyBio Replicators

     
 


The pin tool on the right has 384 pins that are .229 mm in diameter and deliver ~ 2 nl hanging drop and ~4 nl liquid to liquid.

 

 


  1536 Pin Double Float Plate CyBio Replicators

     
 


     


  Replacement Pins for the above 96 and 384 Pin Double Float Plate CyBio Replicators.

 


The Double Float Plate CyBio style Replicators have several advantages over the E-Clip Style Floating Replicators:

  1. The double float plate pins are smaller in diameter and deliver very small volumes (down to 2 nl).
     

  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).

 


  **NEW** Hydrophobic Coated Pin Replicators

     
 


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

 
 
 


Replacement Pins for the above 96, 384 and 1536 Hydrophobic Pin CyBio Replicators.

 

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

 
 
 

  CyBio/Matrix Slot Pin Replicators
     
 


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 s 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. See customer data

 

 


  Hydrophobic Coated Slot Pin Replicators

     
 


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

 
     


  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 1000 nl by changing the width and depth of the slot.  Inquire by phone or e-mail for more information.

     

 
  Replacement Pins

     
 


Replacement Pins for all the replicators on this page can be found on this hyperlink.

     


  Cleaning Pins On Cybio Robot Pin Tools

     
 


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.

 

     


  Pin Tool Accessory Kit

     
 


We highly recommend that you order the CyBio/Matrix Pin Tool Accessory kit at the time you order the pin tool.  This "essentials" accessory kit contains two Low Profile Pin Washing Reservoirs with anti-splash Baffles to dampen the wave action induced by moving the plates on the robot. The VP 540DB Lint Free Blotting System is used to clean the pins efficiently between samples, 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 OK19 Kit you will receive a 5% discount over the sum of the individual items.

     
   

CyBio/Matrix Pin Tool Accessory Kit - VP OK19

Low Profile Pin Wash Reservoirs (2) VP 542
Anti-Splash Baffles for VP 542 (2 sets) VP 542F
Lint Free Blotting System with pad (1 pkg) VP 540DB
Pin Cleaning Solution (1) VP 110
Pin Cleaning Brush (1) VP 425
Safe Pin Tool Storage Station (1) VP 555
 
         

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