How To Choose the Right Robot Pin Tool – Considerations

The Right Pin Tool - Considerations

The most important feature to consider is the transfer volume range. However, there are multiple factors that affect volume transfer. We have conducted experiments on 6 factors to help you select the best pin for your application. Filling out the Pin Tool Questionnaire before reading this blog will help your understanding of the necessary considerations.

  • Pin Diameter More liquid is transferred on a solid pin with a large diameter as a Hanging Drop. See Chart for Range of Transfer in DMSO and Aqueous Solutions.
  • Slot vs Solid Pin and Size More liquid is transferred with a Slot Pin than with a Solid Pin and the larger the slot the more is transferred. See Chart.
  • Pin Withdrawl Speed from the Source Liquid The faster the withdrawal speed the more liquid is transferred. This phenomenon allows you to significantly adjust the volume transferred with your robot’s speed.  See Chart.
  • Volume of Fluid in the Source Plate Maintaining consistent volume in plates using Solid Pins is more important than when using Slot Pins. See Chart.
  • Pin Surface Tension Hydrophobic Coated pins transfer slightly less than uncoated pins. See Chart
  • Surface Tension of the Transferred Liquid Aqueous and DMSO have different surface tensions which can also be changed by adding protein (BSA), DNA, or surfactants like tween 20. See Chart

 The Decision Tree Of Choosing a Robot Pin Tool

Step One – Pick Mounting Plate for your Robot
This choice will allow the Pin Tool to be manipulated by the robot’s pipettor or plate handler.  Which of these robot functions has the greatest range of withdrawal speeds? If you haven’t purchased a robot yet, talked to us about which one would be best suited for your application.
Step Two – Pick Source Plate Transfer Volume
If the volume in the source wells are low or variable select a slot pin because they fill by capillary action and are less affected by the volume in the source well.  If the source well volume is high or constant select a solid pin as they are more economical. If the pin will transfer a liquid to an agar surface, select a solid pin as they are easier to clean off carry-over agar. More liquid is transferred with a Slot Pin than with a Solid Pin and the larger the slot the more is transferred. See Chart. Maintaining consistent volume in plates using Solid Pins is more important than when using Slot Pins.  See Chart.
Step Three – Pick a Desired Pin Transfer Volume
Pick a Pin based on the combined conditions of the volume in the source microplate, the nature of the liquid (Aqueous, DMSO) and its surface tension (BSA, DNA or Tween 20) the diameter and shape of the wells, and the speed range of withdrawal of your robot. See Charts for the Range of Transfer in DMSO and Aqueous Solutions. The faster the withdrawal speed the more liquid is transferred. This phenomenon allows you to significantly adjust the volume transferred with your robot’s speed. See Chart. Select hydrophobic pin coating if the analyte will non-specific bind to stainless steel Hydrophobic Coated pins transfer slightly less than uncoated pins. See Chart.

We also offer sterile and non- sterile 96 and 384 pin disposable polypropylene pin tools that can be manipulated robotically, these each deliver ~ 100 nl.

Step Four -Pick Microplate Height
If a deep well microplate is selected then a “T” tall pin is required to reach the bottom of the wells. Some robots have limited Z heights and can not accommodate “T” pins with their longer exposed length (30 mm) and longer overall length (50.8mm). Those robots must use the shorter “C” pins and then are restricted to only deep well microplates that are at least 2/3 full or they are restricted to only using standard Microplates. See these drawings (Link) for the total overall height of several pin tools which includes the height of the floating fixture and the height of the mounting plate. These drawings are helpful when selecting a robot.
Step Five -Pick a Microplate Format (24, 48, 96, 384, 1536), Floating Fixture, and Registration Accessory
  Registration Accessories to Match Format
Step Six – Pick the Docking Station
Match Docking Station to robotic liquid handler
Step Seven – Pick Wash Reservoir
Depending on the source liquid and the recipient liquids you need to select the appropriate wash systems. Most customers select our running H2O or ETOH Spillway wash reservoirs. We recommend
Step Eight – Pick Cleaning  Supplies
Blotting the pins on lint-free blotting paper after transferring them to the recipient microplate is very important in reducing carry-over liquids into the wash reservoirs.  We recommend also blotting the pins after each wash reservoir.  We also recommend cleaning the pins with our VP 110A pin cleaning solution at the end of each days use, so they are ready for use the next day. We Recommend
Step Nine – Pick a Pin-Drying System
After the final ETOH wash and blot select a pin drying system or let ETOH evaporate naturally over a minute or 2. We recommend using a fan dryer which only takes a few seconds. VP 904A
Step Ten – Still Have Questions?
Please contact one of our knowledgeable scientists, they will be able to guide you through the process.

Hydrophobic Coated vs Non-Coated Pin Transfers

Solid Pin Delivery Data For Aqueous Solutions In 96 Format With Uncoated And /Ah Hydrophobic Coated Pins
PinDescriptionnl TransferredCV%
0.229 mm diameter (FP9)Total PinUncoated7.412.4
Hydrophobic7.465.4
0.229 mm diameter (FP9)Hanging DropUncoatedN/AN/A
Hydrophobic2.093.8
0.457 mm diameter (FP1)Total PinUncoated33.483.2
Hydrophobic28.177.5
0.457 mm diameter (FP1)Hanging DropUncoated16.964.5
Hydrophobic8.510.8
0.787 mm diameter (FP3)Total PinUncoated87.323.9
Hydrophobic77.43.9
0.787 mm diameter (FP3)Hanging DropUncoated48.771.2
Hydrophobic43.059.4
1.19 mm diameter  (VP 409 & VP 386)Total PinUncoated247.222.8
Hydrophobic192.672.6
1.19 mm diameter (VP 409 & VP 386)Hanging DropUncoated76.351.6
Hydrophobic108.42.8
1.58 mm diameter (VP 408 & VP 384)Total PinUncoated273.54.6
Hydrophobic259.253.1
1.58 mm diameter (VP 408 & VP 384)Hanging DropUncoated201.935
Hydrophobic170.047.5

Transfer Of Horseradish Peroxidase In Tris Buffered Saline With Pin Tools

Conclusion

Coating pins will reduce the total amount of liquid transferred and also reduce the amount of non-specific binding to the stainless-steel pins. If the substance you are transferring has high non-specific binding this will be an important factor in selecting your pins.

Slot Pin Delivery Data For Aqueous Solutions In 96 Format With Uncoated And /Ah Hydrophobic Coated Pin
PinDescriptionnl TransferredCV%
0.229 mm diameter (FP9)Total PinUncoated7.412.4
Hydrophobic7.465.4
0.229 mm diameter (FP9)Hanging DropUncoatedN/AN/A
Hydrophobic2.093.8
0.457 mm diameter (FP1)Total PinUncoated33.483.2
Hydrophobic28.177.5
0.457 mm diameter (FP1)Hanging DropUncoated16.964.5
Hydrophobic8.510.8
0.787 mm diameter (FP3)Total PinUncoated87.323.9
Hydrophobic77.43.9
0.787 mm diameter (FP3)Hanging DropUncoated48.771.2
Hydrophobic43.059.4
1.19 mm diameter  (VP 409 & VP 386)Total PinUncoated247.222.8
Hydrophobic192.672.6
1.19 mm diameter (VP 409 & VP 386)Hanging DropUncoated76.351.6
Hydrophobic108.42.8
1.58 mm diameter (VP 408 & VP 384)Total PinUncoated273.54.6
Hydrophobic259.253.1
1.58 mm diameter (VP 408 & VP 384)Hanging DropUncoated201.935
Hydrophobic170.047.5

Transfer Of Horseradish Peroxidase In Tris Buffered Saline With Pin Tools

Conclusion

Although the slots in the pin are a precise volume, the liquid that is transferred is usually more. The reason for this is due to the surface tension of the liquid causing the liquid in the slot to “bow out” thus increasing the volume of the liquid in the slot. If is important for you to transfer exactly a certain volume we can make custom slots to match the surface tension characteristics of your liquid

Liquid Surface Tension

Effect Of DNA Or BSA Concentration On Slot Pin Transfers Of Uncoated And Hydrophobic Coated Pins (FP3CS500)
Solvent/SampleConcentrationCV%nl FITC TransferredCV%nl FITC Transferred
UncoatedUncoatedHydrophobic CoatedHydrophobic Coated
DMSO (-)08.1353.427.5298.72
DMSO + DNA (mg/ml)0.56.6497.216.6435.86
0.259432.494.1391.93
0.1258.9363.640.9344.75
0.06252.3381.862331.68
0.03131.5378.034.4331.71
0.01561.2357.521.4329.03
Tris (-)04.9577.317.2493.53
Tris + DNA (mg/ml)0.54.5540.531.1477.5
0.254.6518.216.1456.75
0.12515.8583.254.1438.82
0.06254.2551.173.1433.69
0.03134.4536.662.3458.37
0.01562.9528.531.2441.1
Tris + BSA (%)45.4462.1311409.27
14452.862.7426.58
0.2511.7456.451.3408.72
0.06251.1445.226.5393.07
0.01563.7462.853.9430.2
0.00391.5493.542.2437.29
0.0012.9504.250.7475.96
Conclusions

1. Increasing the concentration of DNA (sheared salmon sperm) to .25 mg/ml significantly increases the volume of DMSO liquid transferred for both coated and uncoated FP3S500 Slot Pins.
2. Increasing the concentration of DNA does not significantly increase the volume of Tris buffer (aqueous) transferred by both coated and uncoated FP3S500 Slot Pins.
3. Increasing the concentration of BSA (Bovine Serum Albumin) significantly decreases the volume of Tris buffer transferred by both coated and uncoated FP3S500 Slot Pins.
4. Hydrophobic coated FP3S500 Slot Pins transferred less DMSO – DNA and less Tris DNA and less Tris BSA than the uncoated FP3S500 Slot Pins.
5. Both coated and uncoated FP3S500 pins transfer significantly more aqueous solution than DMSO.

Effect Of DNA Or BSA Concentration On Slot Pin Transfers Of Uncoated And Hydrophobic Coated Pins (FP1CS50)
Solvent/SampleConcentrationCV%nl FITC TransferredCV%nl FITC Transferred
UncoatedUncoatedHydrophobic CoatedHydrophobic Coated
DMSO (-)04.249.382.149.31
DMSO + DNA (mg/ml)0.54.951.242.656.79
0.251.750.21.249.53
0.1251.551.272.349.77
0.06252.249.344.148.19
0.03131.249.030.250.23
0.01562.445.91.446.64
Tris (-)02.689.512.991.34
Tris + DNA (mg/ml)0.5777.110.684.62
0.253.982.221.684.89
0.1253.985.42185.08
0.06251.585.362.885.03
0.0313284.52388.19
0.01562.682.922.883.2
Conclusions

1. In contrast to the FP3S500 data, increasing the concentration of DNA to .25 mg/ml does not significantly increase the volume of DMSO liquid transferred for both coated and uncoated FP1S50 Slot Pins.
2. Increasing the concentration of DNA does not significantly increase the volume of Tris buffer (aqueous) transferred by both coated and uncoated FP1S50 Slot Pins.
3. In contrast to the FP3S500 data, FP1S50 coated pins transferred about the same volume of DNA at all concentrations as did uncoated pins.
4. Both coated and uncoated FP1S50 pins transfer significantly more aqueous solution than DMSO.
5. The differences between the FP3S500 and the FP1S50 pin may be due to the different pin diameter’s effect on contact angle and therefore on the “wetting” of the pin. See the diagram on the link to / ah energy system.

PinDescriptionnl TransferredCV%
0.229 mm diameter (FP9)Total PinUncoated7.412.4
Hydrophobic7.465.4
0.229 mm diameter (FP9)Hanging DropUncoatedN/AN/A
Hydrophobic2.093.8
0.457 mm diameter (FP1)Total PinUncoated33.483.2
Hydrophobic28.177.5
0.457 mm diameter (FP1)Hanging DropUncoated16.964.5
Hydrophobic8.510.8
0.787 mm diameter (FP3)Total PinUncoated87.323.9
Hydrophobic77.43.9
0.787 mm diameter (FP3)Hanging DropUncoated48.771.2
Hydrophobic43.059.4
1.19 mm diameter  (VP 409 & VP 386)Total PinUncoated247.222.8
Hydrophobic192.672.6
1.19 mm diameter (VP 409 & VP 386)Hanging DropUncoated76.351.6
Hydrophobic108.42.8
1.58 mm diameter (VP 408 & VP 384)Total PinUncoated273.54.6
Hydrophobic259.253.1
1.58 mm diameter (VP 408 & VP 384)Hanging DropUncoated201.935
Hydrophobic170.047.5

Aqueous Transfer with Solid Pins

Hydrophobic coating pins will reduce the total amount of aqueous HRP liquid transferred and also reduce the amount of non-specific binding to the stainless-steel pins. If the substance you are transferring has high non-specific binding this will be an important factor in selecting your pins.

 

Pin diameter also has an effect on the degree of reduction of liquid transfer with hydrophobic coating as the smaller the diameter the less the reduction of transfer. This is most likely due to the curvature of the pin affecting the wetting contact angle

PinDescriptionnl TransferredCV%
0.457 mm diameter (FP1)6 nl SlotTotal Pin*Uncoated25.610.8
HydrophobicN/AN/A
10 nl SlotTotal Pin*Uncoated23.366.1
Hydrophobic25.856.9
50 nl SlotTotal Pin*Uncoated67.832.5
HydrophobicN/AN/A
0.787 mm diameter (FP3)  100 nl SlotTotal Pin*Uncoated180.327.2
Hydrophobic205.845.5
200 nl SlotTotal Pin*Uncoated277.824.9
Hydrophobic287.33.8
500 nl SlotTotal Pin*Uncoated581.165.2
Hydrophobic555.693

DMSO Transfer with Slot Pins

Hydrophobic coating pins will slightly increase the total amount of DMSO FITC liquid transferred.

PinDescriptionnl TransferredCV%
0.787 mm diameter (FP3)    100 nl Slot Total Pin, Including SlotUncoated195.691.6
Hydrophobic170.22.9
0.787 mm diameter (FP3)  100 nl Slot, Slot OnlyUncoated149.674.9
Hydrophobic129.617.6
0.787 mm diameter (FP3)200 nl Slot Total Pin, Including SlotUncoated269.771.9
Hydrophobic228.6217.1
0.787 mm diameter (FP3)200 nl Slot, Slot OnlyUncoated237.528.9
Hydrophobic186.95.9

Aqueous Transfer with Slot Pins

Although the slots in the pin are a precise volume, the liquid that is transferred is usually more because of the volume carried on the sides of the pins. 

As seen with other aqueous data the amount transferred on hydrophobic coated Slot pins is less than on uncoated Solid or Slot pins. Thus Hydrophobic coating has the most effect on aqueous transfers.

Withdrawl Speeds Impact on Volume Transfer

Solid Pins More affected by Source Plate Volume

Volume Transferred For FP1 Pins (Uncoated) In 96 And 384 Formats
Volume Transferred For FP3 Pins (Uncoated) In 96 And 384 Formats

Note: Same volume (200ul for 96 Format and 74 ul for 384 Format) in recipient plates and same pin withdrawal speed for all pins. Changes to pin withdrawal speed or volume in the source plate can result in different volumes being transferred.

Transfer volumes should always be confirmed by customers for their assay conditions and automated system.

Aqueous Solutions Pin Transfer Volumes Ranges

Aqueous Solutions on Uncoated Pins in 96 Format Microplates(1)
Pin TypePin Diameter(mm)Shape96 Format Low Range(nL)²96 Format High Range(nL)²
FP90.229Solid1339
FP80.356Solid1537
FP10.457Solid2261
FP1S60.4576nL Slot3467
FP1S100.45710nL Slot3974
FP1S500.45750nL Slot90124
FP30.787Solid93213
FP3S1000.787100nL Slot213334
FP3S2000.787200nL Slot311449
FP3S5000.787500nL Slot515671
FP40.914Solid126289
Footnotes: (1) Delivery volume range is determined by speed of withdrawal from source liquid: Z-Speed Range = 1.5-30 mm/sec, slow speed = low volume delivery range, fast speed = high volume delivery range (2) 200ul source plate volume per well
Aqueous Solutions on Hydrophobic Pins in 96 Format Microplates(1)
Pin TypePin Diameter(mm)Shape96 Format Low Range(nL)²96 Format High Range(nL)²
FP90.229Solid1338
FP80.356Solid
FP10.457Solid2360
FP1S60.4576nL Slot3367
FP1S100.45710nL Slot4075
FP1S500.45750nL Slot86119
FP30.787Solid76209
FP3S1000.787100nL Slot188324
FP3S2000.787200nL Slot288436
FP3S5000.787500nL Slot473649
FP40.914Solid
Footnotes: (1) Delivery volume range is determined by speed of withdrawal from source liquid: Z-Speed Range = 1.5-30 mm/sec, slow speed = low volume delivery range, fast speed = high volume delivery range (2) 200ul source plate volume per well
Aqueous Solution on E-Clip, Uncoated Pins(1)
Pin TypePin Diameter(mm)ShapeLow Range(nL)²High Range(nL)²
FP1.58Solid Pointed175594
FPS.51.58500nL Slot524962
FPS1.581000nL Slot10561476
FPS21.582000nL Slot17392174
FPS51.585000nL Slot51504953
FP61.58Solid Flat465960
FP6S.51.58500nL Slot9341445
FP6S1.581000nL Slot13961930
FP6S21.582000nL Slot20722637
FP6S51.585000nL Slot48204693
Footnotes:(1) Delivery volume range is determined by speed of withdrawal from source liquid: Z-Speed Range = 1.5-30 mm/sec, slow speed = low volume delivery range, fast speed = high volume delivery range (2) 200ul source plate volume per well for 96 Format and 75ul source plate volume per well for 384 Format

DMSO Pin Transfer Volume Range Charts

Uncoated Pins in 96 and 384 Format Microplates(1)
Pin TypePin Diameter(mm)Shape96 Format Low Range(nL)²96 Format High Range(nL)²384 Format Low Range(nL)³384 Format High Range(nL)³
FP90.229Solid41038
FP80.35Solid1326618
FP10.457Solid18431131
FP1S60.4576nL Slot24491534
FP1S100.45710nL Slot30542140
FP1S200.45720nL Slot37612746
FP1S300.45730nL Slot46683554
FP1S400.45740nL Slot57784563
FP1S500.45750nL Slot70905675
FP30.787Solid671392979
FP40.91Solid941973498
FP3S1000.787100nL Slot175241114163
FP3S2000.787200nL Slot280332203250
FP3S5000.787500nL Slot535559427464
FP4S10000.911000nL Slot9401011704800
FP4S20000.912000nL Slot1518160812771362
Footnotes: (1) Delivery volume range is determined by speed of withdrawal from source liquid: Z-Speed Range = 1.5-30 mm/sec, slow speed = low volume delivery range, fast speed = high volume delivery range (2) 200ul source plate volume per well (3) 75ul source plate volume per well
Hydrophobic-coated Pins in 96 and 384 Format Microplates(1)
Pin TypePin Diameter (mm)Shape96 Format Low Range(nL)²96 Format High Range(nL)²384 Format Low Range(nL)³384 Format High Range(nL)³
FP9H0.229Solid41038
FP8H0.35Solid924617
FP1H0.457Solid1539927
FP1S6H0.4576nL Slot23491432
FP1S10H0.45710nL Slot29532038
FP1S20H0.45720nL Slot35592643
FP1S30H0.45730nL Slot47693553
FP1S40H0.45740nL Slot54754158
FP1S50H0.45750nL Slot69905773
FP3H0.787Solid671342776
FP4H0.91Solid9518932102
FP3S100H0.787100nL Slot170227108164
FP3S200H0.787200nL Slot266320190239
FP3S500H0.787500nL Slot520542416456
FP4S1000H0.911000nL Slot9321000741805
FP4S2000H0.912000nL Slot1571163813511423
Footnotes: (1) Delivery volume range is determined by speed of withdrawal from source liquid: Z-Speed Range = 1.5-30 mm/sec, slow speed = low volume delivery range, fast speed = high volume delivery range (2) 200ul source plate volume per well (3) 75ul source plate volume per well
E-Clip, Uncoated Pins, for 96 and 384 Format Microplates(1)
Pin TypeDiameter (mm)Shape96 Format Low Range(nL)²96 Format High Range(nL)²384 Format Low Range(nL)³384 Format High Range(nL)³
FP1.58Solid Pointed147411168395
FPS.51.58500nL Slot442704631843
FPS1.581000nL Slot893113013431498
FPS21.582000nL Slot1911203826072767
FPS51.585000nL Slot3908429651805253
FP61.58Solid Flat323674154398
FP6S.51.58500nL Slot73410428551053
FP6S1.581000nL Slot1210150016381717
FP6S21.582000nL Slot2299238427873068
FP6S51.585000nL Slot4329465652375245
Footnotes:(1) Delivery volume range is determined by speed of withdrawal from source liquid: Z-Speed Range = 1.5-30 mm/sec, slow speed = low volume delivery range, fast speed = high volume delivery range (2) 200ul source plate volume per well (3) 75ul source plate volume per well