Patricia Koutz Director of R&D

Patricia KoutzDirector of R&D

Introduction to V&P Scientific Pin Tools and Accessories for

Tecan Workstations

Private and Confidential22

Advantages of V&P Pin ToolsAdvantages of V&P Pin ToolsDelivery from 2 nl to 5 ul.Delivery from 2 nl to 5 ul.

Nanoliter delivery with CVs averaging ~5% Nanoliter delivery with CVs averaging ~5% fluorescein.fluorescein.

Convert a retired 96 robot to 384 or 1536 Convert a retired 96 robot to 384 or 1536 pin tool station.pin tool station.

Accommodates 1536 format easily.Accommodates 1536 format easily.

Nanoliter delivery facilitates assay Nanoliter delivery facilitates assay miniaturization.miniaturization.

Pin tool assay procedure shortens cycle time Pin tool assay procedure shortens cycle time by half.by half.


Advantages of V&P Pin ToolsAdvantages of V&P Pin Tools

Eliminates dilution step in screening assays to Eliminates dilution step in screening assays to save time, reagents and disposable costssave time, reagents and disposable costs. .

(10-50nL direct into assay plate)(10-50nL direct into assay plate)

No disposable tip costs – pays for tool by No disposable tip costs – pays for tool by second run.second run.

Method development is quick.Method development is quick.

Pin tools mount to 45 commercial robot Pin tools mount to 45 commercial robot systems.systems.

Pin tools are useful in many applications.Pin tools are useful in many applications.


Tecan Floating Pin Tool – Tecan Floating Pin Tool – MCA/TeMOMCA/TeMO

Private and Confidential

Tecan Floating Pin Tool – RoMATecan Floating Pin Tool – RoMA

55


What Makes a Pin Tool?What Makes a Pin Tool?

The PinsThe PinsThe Fixture- The Fixture- 2 float plates2 float plates

The Mounting PlateThe Mounting Plate


V&P PinsV&P PinsLarge selection of pin diameters, slots, lengths, and Large selection of pin diameters, slots, lengths, and coatings to accommodate many applications. coatings to accommodate many applications.

Slot width is typically less than 1/3 the diameter of the Slot width is typically less than 1/3 the diameter of the pin.pin.

Solid pin with Solid pin with hanging drophanging drop

Slot pin with liquid in slot


V&P Pin DiametersV&P Pin Diameters


Tube & E-Clip Style PinsTube & E-Clip Style Pins


E-Clip Style PinsE-Clip Style Pins

Common UsesCommon UsesLarger Volume TransfersLarger Volume Transfers

(500nl-5ul)(500nl-5ul)Cell TransfersCell Transfers

(Yeast, E. coli, etc.):(Yeast, E. coli, etc.):• Liquid-to-liquid Liquid-to-liquid (Solid or slotted pins)(Solid or slotted pins)• Liquid-to-membrane Liquid-to-membrane (Solid or slotted pins)(Solid or slotted pins)• Liquid-to-agar Liquid-to-agar (Solid pins only)(Solid pins only)• Agar-to-agar Agar-to-agar (Solid pins only)(Solid pins only)

1.58mm 1.58mm

1.58mm

1.58mm

4.77mm

2.36mm

2.36mm

3.18mm


FP6 Slot PinsFP6 Slot Pins

1.58mm diameter


Tube-Style PinsTube-Style Pins

Common UsesCommon UsesVery Small Volume Transfers (2-500nl)Very Small Volume Transfers (2-500nl)

Compound TransfersCompound Transfers






Misc. InfoMisc. Info

Liquid transfers with slotted pinsLiquid transfers with slotted pins• Slots fill by capillary actionSlots fill by capillary action• Slots do not need to be submerged to fillSlots do not need to be submerged to fill• Slots MUST be submerged to transfer liquidSlots MUST be submerged to transfer liquid

The slot of the FP6S5 pin (5ul volume) is The slot of the FP6S5 pin (5ul volume) is 6mm deep.6mm deep.

Watch liquid displacement in wells of Watch liquid displacement in wells of microplate as pins enter the wells.microplate as pins enter the wells.


V&P Floating Pin Tool FixtureV&P Floating Pin Tool Fixture


V&P Mono Pin Tool FixtureV&P Mono Pin Tool Fixture

Mono Pin Tool on MWG RoboAmpMono Pin Tool on MWG RoboAmp


V&P Rotational Mounting Plate V&P Rotational Mounting Plate for Tecan MCA/TeMO Pin Toolsfor Tecan MCA/TeMO Pin Tools

1818


V&P Mounting Plate for Tecan V&P Mounting Plate for Tecan RoMA Pin ToolsRoMA Pin Tools

1919


Guide for SelectingGuide for Selectingthe Right Pin Toolthe Right Pin Tool

1.1. Robot system.Robot system.

2.2. Volume to be transferred.Volume to be transferred.

3.3. Volume in source plate.Volume in source plate.

4.4. Solid pins versus slotted pins.Solid pins versus slotted pins.

5.5. Non-specific binding to stainless Non-specific binding to stainless steel.steel.

6.6. ““Z” clearance on robot deck.Z” clearance on robot deck.



1.1. Robot system.Robot system.• V&P RoMA Mounting Plate (BMPROMA)V&P RoMA Mounting Plate (BMPROMA)

• V&P standard MCA/TeMO mounting plate (V&P standard MCA/TeMO mounting plate (BMPTECANBMPTECAN))

• V&P rotational MCA/TeMO mounting plate (BMPTECANR)V&P rotational MCA/TeMO mounting plate (BMPTECANR)




5.5. Non-specific binding to stainless steel.Non-specific binding to stainless steel.



Tecan TeMO Pin ToolTecan TeMO Pin Tool(Yale)(Yale)


Tecan RoMA Pin Tool Tecan RoMA Pin Tool (U of Kentucky)(U of Kentucky)

2323


V&P Docking Station for V&P Docking Station for Tecan Pin ToolsTecan Pin Tools


V&P Docking Station for V&P Docking Station for Tecan Pin ToolsTecan Pin Tools










Factors Affecting Pin Delivery Factors Affecting Pin Delivery VolumeVolume

Pin diameter.Pin diameter.

Volume of slot in pin.Volume of slot in pin.

Surface tension of the liquid being transferred.Surface tension of the liquid being transferred.

Surface tension of the pin. – how clean? Surface tension of the pin. – how clean? Uneven transfers due to the sides and slot volume variationUneven transfers due to the sides and slot volume variation

Depth to which the pin is submerged in source Depth to which the pin is submerged in source plate.plate.

SPEED OF REMOVAL OF PIN FROM SOURCE SPEED OF REMOVAL OF PIN FROM SOURCE LIQUID.LIQUID.


Speed of removal of pinsSpeed of removal of pins

Pull pins out SLOW from source platesPull pins out SLOW from source plates• Excess liquid will wick off and LESS volume will Excess liquid will wick off and LESS volume will

be transferredbe transferred

Pull pins out FASTPull pins out FAST• Excess liquid does Excess liquid does notnot wick off and HIGHER wick off and HIGHER

volumes will be transferred volumes will be transferred

INCREASE SPEED = INCREASE VOLUMEINCREASE SPEED = INCREASE VOLUME


Speed of removal of pinsSpeed of removal of pins

During Customer Set Up:During Customer Set Up:

Customers set up a standard curve and Customers set up a standard curve and mock fluorescein assay at various mock fluorescein assay at various withdrawal speeds to determine the withdrawal speeds to determine the appropriate speed for the volume desired.appropriate speed for the volume desired.

CV’s generally do not change with speed.CV’s generally do not change with speed.


V&P Scientific Nanoliter PinsV&P Scientific Nanoliter Pins

PINPIN DIAMETERDIAMETER SHAPESHAPE DELIVERYDELIVERY

VOLUMEVOLUME

RANGERANGE1,21,2

FP9FP9 0.229 mm0.229 mm SolidSolid 4 - 10 nl4 - 10 nl


FP1S10FP1S10 0.457 mm0.457 mm 10 nl Slot10 nl Slot 29 - 65 nl29 - 65 nl




FP3S200FP3S200 0.787 mm0.787 mm 200 nl Slot200 nl Slot 284 - 401nl284 - 401nl

FP3S500FP3S500 0.787 mm0.787 mm 500 nl Slot500 nl Slot 505 - 583 nl505 - 583 nl1 Volume range determined by speed of withdrawal from source plate2 Change in speed 7-fold, liquid transferred in DMSO


What happens exactly?What happens exactly?

Pin tools are a TRANSFER tool, not a Pin tools are a TRANSFER tool, not a dispensing tool.dispensing tool.• Example: If you transfer 5 ul from the source plate to Example: If you transfer 5 ul from the source plate to

45 ul in the destination plate with a pin tool, you will 45 ul in the destination plate with a pin tool, you will have 50 ul of a 1:10 dilution in the destination plate…have 50 ul of a 1:10 dilution in the destination plate…while the pins are in the wells.while the pins are in the wells.

• When the pins are withdrawn from the wells, they will When the pins are withdrawn from the wells, they will remove 5 ul of the 1:10 dilution, leaving 45 ul final remove 5 ul of the 1:10 dilution, leaving 45 ul final volume in the destination plate. volume in the destination plate.

Solid pins transfer liquid on the sides of the pins Solid pins transfer liquid on the sides of the pins and larger hanging dropletand larger hanging dropletSlotted pins transfer liquid on the sides, in the Slotted pins transfer liquid on the sides, in the slot and hanging dropletslot and hanging droplet


What happens exactly?What happens exactly?

Pre-wet pins in source platePre-wet pins in source plate• Move pins up and down through the meniscus, 3X Move pins up and down through the meniscus, 3X

(completely out of the liquid).(completely out of the liquid).• If the pins hit a air bubble going into well, this mixing If the pins hit a air bubble going into well, this mixing

will displace it.will displace it.• Similar to pre wetting pipetting tips.Similar to pre wetting pipetting tips.

Post-transfer mix in destination platePost-transfer mix in destination plate• Move pins up and down through the meniscus, 3X. Move pins up and down through the meniscus, 3X. • Pins act as piston moving up and down to mix. Pins act as piston moving up and down to mix. • Just letting pins sit in liquid doesn’t help increase Just letting pins sit in liquid doesn’t help increase

amount of volume transfer.amount of volume transfer.


Last Withdrawal SpeedLast Withdrawal Speed

The LAST mix withdrawal of the pins from The LAST mix withdrawal of the pins from the source plate is the most important.the source plate is the most important.This speed is determined from the mock This speed is determined from the mock fluorescein assay run during set up.fluorescein assay run during set up.Use the same speed and number of cycles Use the same speed and number of cycles in source and destination plates so the in source and destination plates so the transfer amounts will be the same.transfer amounts will be the same.The first two up and down mix movements The first two up and down mix movements can be faster.can be faster.










Affect of Variable Source Plate Affect of Variable Source Plate Volumes on Pin Transfer VolumeVolumes on Pin Transfer Volume

0%

10%

20%

30%

40%

50%

60%

70%

80%

90%

100%

FP

6

FP

S.5

FP

6S.5

FP

S

FP

6S

FP

S2

FP

6S2

FP

S5

FP

6S5

FP

4

FP

Pins

% T

ran

sfer

red

200 ul100 ul50 ul

Volume in Source Well






4.4. Solid pins versus slotted pins.Solid pins versus slotted pins.• Cost, CV’sCost, CV’s




Hydrophobic/Lipophobic CoatingHydrophobic/Lipophobic Coating

Prevents or reduces non-specific binding.Prevents or reduces non-specific binding.

Prevents or reduces lipid and lipoprotein binding.Prevents or reduces lipid and lipoprotein binding.

Prevents liquid “creep” up the pinPrevents liquid “creep” up the pin..

Increases size of hanging drop.Increases size of hanging drop.

Decreases volume on side of pin.Decreases volume on side of pin.

Coating can be applied to all V&P stainless steel Coating can be applied to all V&P stainless steel pins. pins.

At least $2 more vs. uncoated pin.At least $2 more vs. uncoated pin.

Whole pin coated including slot.Whole pin coated including slot.








6.6. ““Z” clearance on robot deck.Z” clearance on robot deck.• Long pins versus short pinsLong pins versus short pins


Tube-Style Exposed Pin LengthTube-Style Exposed Pin Length


Questions To Ask CustomersQuestions To Ask Customers

What is the pin tool format?What is the pin tool format?• 96, 384 or 1536 pins?96, 384 or 1536 pins?

What are the mother and daughter plate formats?What are the mother and daughter plate formats?• 96, 384 or 1536?96, 384 or 1536?• Standard microplates or deep well plates?Standard microplates or deep well plates?

What is the typical volume in the mother and daughter What is the typical volume in the mother and daughter plates?plates?• For example: For example:

• Average of 50ul liquid in the 384 well mother plateAverage of 50ul liquid in the 384 well mother plate

• Average of 200ul liquid in the 96 well daughter plate or Average of 200ul liquid in the 96 well daughter plate or

agar surface in Nunc Omni Tray for daughter plate.agar surface in Nunc Omni Tray for daughter plate.


Questions To Ask CustomersQuestions To Ask Customers(Cont.)(Cont.)

What volume do you need to transfer between the What volume do you need to transfer between the mother and daughter plate?mother and daughter plate?

What type of sample is being transferred?What type of sample is being transferred? • For example: For example:

• Transferring compound libraries in 100% DMSO from Transferring compound libraries in 100% DMSO from mother plate to aqueous cell-based assays in daughter mother plate to aqueous cell-based assays in daughter plates. plates.

• Transferring bacterial or yeast cells in culture media Transferring bacterial or yeast cells in culture media from mother plate to agar surface in daughter plate.from mother plate to agar surface in daughter plate.

Will the pin tool be attached to the instrument head or be Will the pin tool be attached to the instrument head or be picked up with the gripper arm?picked up with the gripper arm?


Pin Tool DataPin Tool Data


Are your pins clean?Are your pins clean?


Pin Validation AssayPin Validation Assay (Fluorescein Transfer)(Fluorescein Transfer)

MaterialsMaterialsV&P Scientific pin tools.V&P Scientific pin tools.

CyBi™-Well 96 Liquid Handling Robot.CyBi™-Well 96 Liquid Handling Robot.

Fluorescein Isothiocyanate (FITC), Isomer 1. Fluorescein Isothiocyanate (FITC), Isomer 1. Sigma P/N F7250.Sigma P/N F7250.

100% DMSO. Sigma P/N D2650.100% DMSO. Sigma P/N D2650.

100 mM Tris, pH 8.0. Sigma P/N T3038.100 mM Tris, pH 8.0. Sigma P/N T3038.

Microfluor 96-Well Microtiter Plates, Dynex Microfluor 96-Well Microtiter Plates, Dynex Technologies P/N 7805.Technologies P/N 7805.

PerkinElmer/Wallac VICTOR2 Multilabel Reader.PerkinElmer/Wallac VICTOR2 Multilabel Reader.


Pin Validation AssayPin Validation Assay MethodsMethods

Dip pin tool into source plate (3X)Dip pin tool into source plate (3X)

Pins removed from source plate at controlled Pins removed from source plate at controlled speedsspeeds

Dip pin tool into assay plate (3X)Dip pin tool into assay plate (3X)

Wash pinsWash pins

RepeatRepeat

Withdrawal speeds: 0.78-5.76 cm/secWithdrawal speeds: 0.78-5.76 cm/sec



Pin Tool MaintenancePin Tool Maintenance

Daily cleaning – Before and after assay.Daily cleaning – Before and after assay.Cleaning in-between sample transfers.Cleaning in-between sample transfers.Heavy duty cleaning.Heavy duty cleaning.• Frequently used pin tools - clean once a weekFrequently used pin tools - clean once a week• Infrequently used pin tools - clean once a monthInfrequently used pin tools - clean once a month


VP 540DB Blotting StationVP 540DB Blotting Station


VP 110 Pin Cleaning SolutionVP 110 Pin Cleaning Solution

Available in 30ml bottles of Available in 30ml bottles of 5X concentrated solution.5X concentrated solution.

Dilute 1:5 in distilled water.Dilute 1:5 in distilled water.

Dilution can be reused up to Dilution can be reused up to two months if stored in two months if stored in sealed container.sealed container.


Cleaning Pins In-BetweenCleaning Pins In-BetweenSample Transfers (cont.)Sample Transfers (cont.)

Thorough washing uses 7 deck positions totalThorough washing uses 7 deck positions total96 & 384 pin tools can be blotted in offset 96 & 384 pin tools can be blotted in offset positions in VP 540DB Blotting Station.positions in VP 540DB Blotting Station.• ~2.25-4.5mm offset.~2.25-4.5mm offset.

Blot in same position on Blotting Station for Blot in same position on Blotting Station for each wash station.each wash station.Watch how high the pins are pushed up into Watch how high the pins are pushed up into the float plate during blotting.the float plate during blotting. If excess liquid is not blotted from slots, the If excess liquid is not blotted from slots, the pins may take 60-90 sec to dry unless a fan or pins may take 60-90 sec to dry unless a fan or vacuum system is used.vacuum system is used.


Cleaning Pins In-BetweenCleaning Pins In-BetweenSample Transfers (cont.)Sample Transfers (cont.)

Link to Technical Note for liquid sample Link to Technical Note for liquid sample transfer with robot pin tools and cleaning transfer with robot pin tools and cleaning robot pin toolsrobot pin tools::

http://www.vp-scientific.com/Technotes/135.pdf

Pin Tool Pin Tool AccessoriesAccessories


Converting Manual to Robotic Converting Manual to Robotic Pin ToolPin Tool

Tube-style (and some E-Clip-style) Manual Tube-style (and some E-Clip-style) Manual V&P Pin Tools can be converted to Tecan-V&P Pin Tools can be converted to Tecan-compatible robotic Pin Tool by removal of the compatible robotic Pin Tool by removal of the Basic Guide Pin Plate (BGP).Basic Guide Pin Plate (BGP).


Tecan Pin Tool Docking StationTecan Pin Tool Docking Station

VP 550A Docking Station and TeMO Adaptor Plate


MovieMovie

5555

TeMO Picking Up Pin Tool TeMO Picking Up Pin Tool from Docking Stationfrom Docking Station

Movie Pickup PinTool in Dock of TeMo.mpg


MovieMovie

5656

TeMO Setting Pin Tool TeMO Setting Pin Tool into Docking Stationinto Docking Station

Movie Drop PinTool in Dock of TeMo.mpg


Spillway Wash ReservoirsSpillway Wash Reservoirs

VP 549EtOH Wash ReservoirVP 549EtOH Wash Reservoir

VP 549H2O Wash ReservoirVP 549H2O Wash Reservoir


VP 540T Flowing Wash ReservoirsVP 540T Flowing Wash Reservoirs


VP 549PE-EP3 Wash StationVP 549PE-EP3 Wash Station


VP 546B 384 Fountain Wash StationVP 546B 384 Fountain Wash Station


VP 546C 384 Fountain Wash StationVP 546C 384 Fountain Wash Station

Constructed with Constructed with hydrophilic coated hydrophilic coated aluminum.aluminum.Large channel diameter Large channel diameter (2.25mm diameter) (2.25mm diameter) to to accommodate most pipet accommodate most pipet tips in addition to pin tools.tips in addition to pin tools. Reduced channel inlet Reduced channel inlet orifices orifices (0.35mm) (0.35mm) for more for more uniform flow with all wash uniform flow with all wash liquids.liquids.


Disposable Pin ToolsDisposable Pin Tools


New Pin Tool DevelopmentsNew Pin Tool Developments

3456 Pin Tools.3456 Pin Tools.

11stst 3456 pin tool currently in development 3456 pin tool currently in development phase.phase.

Documents

Patricia Koutz Director of R&D