Pin Tool Data

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Pin Tool Data

The following data on the various pins can be extrapolated to all the robot systems and manual systems as the only difference between them is the length of the pin. The critical elements of pin diameter, shape and coating are all the same. As long as these elements remain fixed the CV's will average under 5%.

We and our Customers have conducted many experiments to elucidate the characteristics of transferring liquids with pin tools. We realize that there is probably too much data here and it may be confusing, but better too much than too little. If you have trouble finding the data you need give us a call and we will be happy to help.

If you want to conduct your own assays to quantitate pin transfers, we have prepared some comphrensive methods and procedures you will find helpful. See Technical note 65.

The following is a U-Tube video from the Broad Institute illustrating a cell-based Screen at the Broad Institute using a V&P 384 Pin Tool on High Res Robot:

This video illustrates how our pin tools are used in the field.

Mechanical/Speed Effects on Pin Transfers

Effect of solvent (Aqueous versus DMSO) on volume transferred by solid and slot pins.
Effect of speed of withdrawal on volume transferred by solid and slot pins. See this page also.
Spotting oligo DNA on to dry plates.
Effect of dwell time on dry plate on volume transferred. See this page also.
Effect of volume in source plate on volume transferred.
Effect of concentration of DNA and BSA on volume transferred with uncoated and hydrophobic coated slot pins - liquid to liquid.
Effect of concentration of DNA and BSA on volume transferred with uncoated and hydrophobic coated slot pins - liquid to dry plate.
liquid to Dry plate transfers
Effect of volume and concentration in source plate on volume transferred.
Effect of Pin cleaning using wash and blot technique. Carry Over data - 100 plate transfers on a- Robot pin tool
Effect of Pin cleaning using wash and blot technique. Accuracy data 25 plate transfers with- Manual pin tool..
Effect of Hydrophobic/Lipophobic coating of pins on aqueous transfer.
Effect of Hydrophobic/lipophobic coating of pins on DMSO transfer. See this page also.
Determination of volume in hanging drop versus the sides of the pin. See this page also.
Determination of volume in slot versus the sides of the pin.
Determination of volume transferred and CVs by a manual disposable pin tool

Uncoated and Hydrophobic Coated Solid Pins

Aqueous Solid Pin Delivery Data On Uncoated Pins And Pins With /ah Hydrophobic Coating

Transfer Of Horseradish Peroxidase In Tris Buffered Saline With Pin Tools
Pin	Description		nl Transferred	CV%
0.229 mm diameter (FP9)	Total Pin	Uncoated	7.41	2.4%
0.229 mm diameter (FP9)	Total Pin	Hydrophobic	7.46	5.4%
0.229 mm diameter (FP9)	Hanging Drop	Uncoated	N/A	N/A
0.229 mm diameter (FP9)	Hanging Drop	Hydrophobic	2.09	3.8%
0.457 mm diameter (FP1)	Total Pin	Uncoated	33.48	3.2%
0.457 mm diameter (FP1)	Total Pin	Hydrophobic	28.17	7.5%
0.457 mm diameter (FP1)	Hanging Drop	Uncoated	16.96	4.5%
0.457 mm diameter (FP1)	Hanging Drop	Hydrophobic	8.51	0.8%
0.787 mm diameter (FP3)	Total Pin	Uncoated	87.32	3.9%
0.787 mm diameter (FP3)	Total Pin	Hydrophobic	77.40	3.9%
0.787 mm diameter (FP3)	Hanging Drop	Uncoated	48.77	1.2%
0.787 mm diameter (FP3)	Hanging Drop	Hydrophobic	43.05	9.4%
1.19 mm diameter VP 409 & VP 386	Total Pin	Uncoated	247.22	2.8%
1.19 mm diameter VP 409 & VP 386	Total Pin	Hydrophobic	192.67	2.6%
1.19 mm diameter VP 409 & VP 386	Hanging Drop	Uncoated	76.35	1.6%
1.19 mm diameter VP 409 & VP 386	Hanging Drop	Hydrophobic	108.40	2.8%
1.58 mm diameter VP 408 & VP 384	Total Pin	Uncoated	273.50	4.6%
1.58 mm diameter VP 408 & VP 384	Total Pin	Hydrophobic	259.25	3.1%
1.58 mm diameter VP 408 & VP 384	Hanging Drop	Uncoated	201.93	5.0%
1.58 mm diameter VP 408 & VP 384	Hanging Drop	Hydrophobic	170.04	7.5%

*Total pin = all the liquid on sides of pin plus the hanging drop that is transferred to a liquid filled well.
** Hanging drop = only the liquid in the hanging drop that is transferred to a membrane.

Coating pins will reduce the total amount 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.

DMSO Solid Pin Delivery Data On Uncoated Pins And Pins With /ah Hydrophobic Coating

Transfer of Fluorescein 5-Isothiocyanate-Isomer I (FITC) In 100% DMSO To 1 M Tris, pH 8.0
Pin	Description		nl Transferred	CV%
0.229 mm diameter (FP9)	Total Pin*	Uncoated	3.47	1.6%
0.229 mm diameter (FP9)	Total Pin*	Hydrophobic	3.61	6.8%
0.229 mm diameter (FP9)	Hanging Drop**	Uncoated	0.90	5.6%
0.229 mm diameter (FP9)	Hanging Drop**	Hydrophobic	1.29	8.8%
0.457 mm diameter (FP1)	Total Pin	Uncoated	16.58	3.9%
0.457 mm diameter (FP1)	Total Pin	Hydrophobic	16.06	2.9%
0.457 mm diameter (FP1)	Hanging Drop	Uncoated	3.74	1.9%
0.457 mm diameter (FP1)	Hanging Drop	Hydrophobic	0.66	7.2%
0.787 mm diameter (FP3)	Total Pin	Uncoated	71.11	4.1%
0.787 mm diameter (FP3)	Total Pin	Hydrophobic	N/A	N/A
0.787 mm diameter (FP3)	Hanging Drop	Uncoated	50.8	12.1
0.787 mm diameter (FP3)	Hanging Drop	Hydrophobic	N/A	N/A

*Total pin = volume of DMSO in hanging drop and on sides of pin.
**Hanging drop = volume of DMSO in hanging drop that is transferred to a membrane.

Detailed Assay Method:
To Determine Volumetric Liquid Transfer With Pin Tools Using FITC In 100% DMSO
To Determine Volumetric Liquid Transfer With Pin Tools Using HRP in TBS/Tween 20

Slot Pin Data

Summary Of Pin To Pin Delivery Precision

Pin Diameter	Volume of Slot	Mother well solvent	Recipient well solvent	Label	CV%
1.58 mm	5 ul	Aqueous	Aqueous	Methyl Orange	3.7%
1.58 mm	2 ul	Aqueous	Aqueous	Methyl Orange	4.9%
.457 mm	50 nl	Aqueous	DMSO	FITC	3.2%
.457 mm	50 nl	ETOH	DMSO	FITC	3.4%
.457 mm	10 nl	Aqueous	DMSO	FITC	4.0%
.457 mm	10 nl	ETOH	DMSO	FITC	2.7%
.787 mm	100 nl	Aqueous	Aqueous	Horseradish peroxidase	1.6%
.787 mm	200 nl	Aqueous	Aqueous	Horseradish peroxidase	1.9%

Hydrophobic Slot Pin Data

Aqueous Slot Pin Delivery Data On Uncoated Pins And Pins With /ah Hydrophobic Coating

Transfer Of Horseradish Peroxidase In Tris Buffered Saline With 96 Pin Tools
Pin	Description		nl Transferred	CV%
0.787 mm diameter (FP3)	100 nl Slot Total Pin Including Slot	Uncoated	195.69	1.6%
0.787 mm diameter (FP3)	100 nl Slot Total Pin Including Slot	Hydrophobic	170.20	2.9%
0.787 mm diameter (FP3)	100 nl Slot Slot Only	Uncoated	149.67	4.9%
0.787 mm diameter (FP3)	100 nl Slot Slot Only	Hydrophobic	129.61	7.6%
0.787 mm diameter (FP3)	200 nl Slot Total Pin Including Slot	Uncoated	269.77	1.9%
0.787 mm diameter (FP3)	200 nl Slot Total Pin Including Slot	Hydrophobic	228.62	17.1%
0.787 mm diameter (FP3)	200 nl Slot Slot Only	Uncoated	237.52	8.9%
0.787 mm diameter (FP3)	200 nl Slot Slot Only	Hydrophobic	186.90	5.9%

*Total pin = all the liquid on sides of pin plus the slot that is transferred to a liquid filled well.
** Slot only = only the liquid in the slot that is transferred to a membrane.

Note that in the data above, 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

DMSO Slot Pin Delivery Data On Uncoated Pins And Pins With /ah Hydrophobic Coating

Transfer of Fluorescein 5-Isothiocyanate-Isomer I (FITC) In 100% DMSO To 1 M Tris, pH 8.0
Pin	Description			nl Transferred	CV%
0.457 mm diameter (FP1)	6 nl Slot	Total Pin*	Uncoated	25.6	10.8%
	6 nl Slot	Total Pin*	Hydrophobic	N/A	N/A
	10 nl Slot	Total Pin*	Uncoated	23.36	6.1%
	10 nl Slot	Total Pin*	Hydrophobic	25.85	6.9%
	50 nl Slot	Total Pin*	Uncoated	67.83	2.5%
	50 nl Slot	Total Pin*	Hydrophobic	N/A	N/A
0.787 mm diameter (FP3)	100 nl Slot	Total Pin*	Uncoated	180.32	7.2%
	100 nl Slot	Total Pin*	Hydrophobic	205.84	5.5%
	200 nl Slot	Total Pin*	Uncoated	277.82	4.9%
	200 nl Slot	Total Pin*	Hydrophobic	287.30	3.8%
	500 nl Slot	Total Pin*	Uncoated	581.16	5.2%
	500 nl Slot	Total Pin*	Hydrophobic	555.69	3.0%

Total pin* = volume of DMSO in hanging drop and on sides of pin.

For customer slot pin tool data please visit this hyperlink

Effect of Source Plate Volume on Pin Volume Transferred

Because the pins carry liquid on the sides as well as the tips and in slots, the volume transferred by pins will be affected by the depth the pin is submerged into the source plate. The following experiment addresses this phenomenon. The results illustrate the importance of having the same volume in all the source wells when using solid pins. This differs from slot pin transfers that are not significantly affected by volume. In that case, the larger the slot the less affect source plate volume has on the transfer volume.

Assay conditions:

Hydrophobic coated and uncoated, solid and slot pins were dipped into varied volumes of FITC in 100% DMSO or 0.1M Tris Buffer in 96 well source plates (25, 50, 100, and 200 ul).

Pins were removed from source plate at a speed of 0.78 cm/sec.

Pins then transferred the FITC to 200 ul 0.1 M Tris, pH 8.0 in black polystyrene assay plate. The fluorescence was measured in a Victor Plate Reader from which the volume of FITC transferred was calculated.

In the following table the volume transferred from the 200 ul source plate was assigned the value of 100% and the volumes transferred by the same pins from source plates with 100, 50 and 25 ul volumes were calculated as a percentage of this volume.

DMSO Transfers - Uncoated Pins
		Source Plate Volume
Pin	Coating	200 ul	100 ul	50 ul	25 ul
FP9	U	100.0%	70.9%	49.4%	28.1%
FP8	U	100.0%	64.9%	41.6%	N/A
FP1	U	100.0%	71.3%	53.5%	37.0%
FP1S6	U	100.0%	80.3%	66.8%	N/A
FP1S10	U	100.0%	83.5%	71.2%	N/A
FP1S20	U	100.0%	86.9%	77.2%	N/A
FP1S50	U	100.0%	89.3%	83.4%	N/A
FP3	U	100.0%	82.5%	66.9%	63.9%
FP3S100	U	100.0%	93.1%	83.8%	84.6%
FP3S200	U	100.0%	95.8%	89.6%	91.0%
FP6	U	100.0%	81.7%	68.9%	78.7%
FP3S500	U	100.0%	98.5%	93.0%	94.9%
FPS.5	U	100.0%	90.4%	77.1%	81.4%
FP6S.5	U	100.0%	90.3%	78.6%	86.5%
FPS	U	100.0%	94.0%	90.4%	85.5%
FP6S	U	100.0%	94.6%	89.8%	83.0%
FPS2	U	100.0%	96.1%	93.5%	90.3%
FP6S2	U	100.0%	96.5%	94.2%	85.4%
FPS5	U	100.0%	97.0%	95.3%	87.5%
FP6S5	U	100.0%	93.1%	94.8%	89.8%
FP4	U	100.0%	61.4%	49.8%	24.3%
FP	U	100.0%	69.9%	50.6%	50.3%

Aqueous Transfers - Uncoated Pins
		Source Plate Volume
Pin	Coating	200 ul	100 ul	50 ul	25 ul
FP9	U	100.0%	79.4%	52.4%	36.0%
FP8	U	100.0%	67.9%	50.8%	43.4%
FP1	U	100.0%	77.2%	68.1%	61.0%
FP1S6	U	100.0%	79.0%	66.6%	N/A
FP1S10	U	100.0%	81.9%	71.9%	N/A
FP1S20	U	100.0%	87.9%	82.5%	N/A
FP1S50	U	100.0%	94.4%	89.8%	N/A
FP3	U	100.0%	92.0%	87.6%	87.7%
FP3S100	U	100.0%	98.3%	93.2%	94.3%
FP3S200	U	100.0%	99.7%	97.7%	100.5%
FP6	U	100.0%	99.6%	96.4%	N/A
FP3S500	U	100.0%	108.8%	109.8%	N/A
FPS.5	U	100.0%	92.6%	88.0%	N/A
FP6S.5	U	100.0%	100.1%	96.2%	N/A
FPS	U	100.0%	99.7%	95.3%	N/A
FP6S	U	100.0%	101.6%	98.8%	N/A
FPS2	U	100.0%	99.7%	96.0%	N/A
FP6S2	U	100.0%	100.0%	98.5%	N/A
FPS5	U	100.0%	98.8%	95.3%	N/A
FP6S5	U	100.0%	98.4%	98.4%	N/A
FP4	U	100.0%	82.2%	74.8%	N/A
FP	U	100.0%	63.8%	47.4%	N/A

DMSO Transfers - Hydrophobic Pins
		Source Plate Volume
Pin	Coating	200 ul	100 ul	50 ul	25 ul
FP9	H	100.0%	66.2%	47.2%	28.8%
FP1	H	100.0%	70.7%	54.1%	37.7%
FP1S6	H	100.0%	78.3%	64.4%	N/A
FP1S10	H	100.0%	81.0%	69.1%	N/A
FP1S20	H	100.0%	85.4%	76.1%	N/A
FP1S30	H	100.0%	87.5%	78.6%	N/A
FP1S40	H	100.0%	87.6%	79.3%	N/A
FP1S50	H	100.0%	89.8%	82.6%	N/A
FP3	H	100.0%	89.2%	75.8%	73.3%
FP3S100	H	100.0%	93.1%	84.6%	84.1%
FP3S200	H	100.0%	95.3%	88.8%	91.1%
FP3S500	H	100.0%	100.3%	93.7%	95.6%

Aqueous Transfers - Hydrophobic Pins
		Source Plate Volume
Pin	Coating	200 ul	100 ul	50 ul	25 ul
FP9	H	100.0%	75.1%	49.1%	32.8%
FP1	H	100.0%	75.6%	66.8%	59.3%
FP1S6	H	100.0%	77.9%	65.6%	N/A
FP1S10	H	100.0%	83.9%	74.0%	N/A
FP1S20	H	100.0%	86.8%	77.9%	N/A
FP1S30	H	100.0%	88.0%	83.0%	N/A
FP1S40	H	100.0%	90.8%	84.4%	N/A
FP1S50	H	100.0%	92.7%	89.1%	65.5%
FP3	H	100.0%	98.4%	93.0%	95.0%
FP3S100	H	100.0%	99.8%	96.0%	97.9%
FP3S200	H	100.0%	101.5%	99.7%	N/A
FP3S500	H	100.0%	107.5%	111.1%	N/A

Data for a similar volume of source plate well experiment, where the concentration was increased proportional to the volume drop (edge effect drying) is on this page.

Accuracy and Precision Data on FP3 Pins
Twenty-five Manual FP3 Replicate Transfers

Assay conditions:

FP3 Pins on 96 pin tool dipped into 200 ul of 0.6 mg/ml FITC in 100% DMSO in 96 well source plate.

Pins removed from source plate at slow speed by hand.

Pins transferred to 200 ul 0.1 M Tris, pH 8.0 in black polystyrene assay plate.

Pins washed with DMSO and Isopropanol and process repeated 25 times.

Standard Deviations of Each Individual Pin Over Twenty five Transfers
12210	10367	10889	12027	12556	12204	12077	12797	12838	13240	14214	12837
9870	9101	8968	9045	10444	11369	12941	12231	11566	13185	13072	12412
10879	10534	7916	9149	9823	9579	11002	11583	11641	12318	12417	12836
12507	9917	8424	9013	10445	10474	10695	11756	10618	11671	12609	14009
10316	10692	8672	8943	10516	9939	11731	12823	11959	14614	12736	12803
12427	11942	10489	11892	11121	11015	10979	12379	11437	12977	12022	13289
11469	12009	10463	11143	11483	12090	11245	11206	10163	11498	11609	11974
11928	13380	12297	13092	12249	10459	14758	12768	12807	12946	11533	13598

Means of Each Individual Pin Over Twenty five Transfers
175199	167987	168780	179896	177477	172754	176475	178194	173510	180418	173092	175342
172809	166962	168435	171034	168005	168666	181265	169233	168903	173762	172993	169765
167298	165832	169438	175892	169829	178468	172546	174387	177447	171384	169894	169421
169084	167889	173436	181685	176511	170238	179931	179247	170418	173792	172997	179068
165882	176052	166423	168458	168968	174238	179874	173538	181658	179024	175375	169496
165960	173950	174790	182885	174083	175978	183721	181238	169725	181726	171937	167683
171445	167841	166939	164066	175782	170673	170084	178250	168350	177029	166365	171713
163360	170203	170746	164877	165561	166233	175879	168842	175449	172994	179367	180043

CV% of Each Individual Pin Over Twenty five Transfers
7.0%	6.2%	6.5%	6.7%	7.1%	7.1%	6.8%	7.2%	7.4%	7.3%	8.2%	7.3%
5.7%	5.5%	5.3%	5.3%	6.2%	6.7%	7.1%	7.2%	6.8%	7.6%	7.6%	7.3%
6.5%	6.4%	4.7%	5.2%	5.8%	5.4%	6.4%	6.6%	6.6%	7.2%	7.3%	7.6%
7.4%	5.9%	4.9%	5.0%	5.9%	6.2%	5.9%	6.6%	6.2%	6.7%	7.3%	7.8%
6.2%	6.1%	5.2%	5.3%	6.2%	5.7%	6.5%	7.4%	6.6%	8.2%	7.3%	7.6%
7.5%	6.9%	6.0%	6.5%	6.4%	6.3%	6.0%	6.8%	6.7%	7.1%	7.0%	7.9%
6.7%	7.2%	6.3%	6.8%	6.5%	7.1%	6.6%	6.3%	6.0%	6.5%	7.0%	7.0%
7.3%	7.9%	7.2%	7.9%	7.4%	6.3%	8.4%	7.6%	7.3%	7.5%	6.4%	7.6%

All Plates, Manual transfers
Std Dev.	12429
Mean	172832
Maximum	216038
Minimum	140955
CV%	7.2%
mg FITC/well	0.0293
nl FITC Transferred	73.299

Extensive raw data and graphs of this experiment are found at this hyperlink.

Determination of Volume of Hanging Drop on FP3 Pin

Assay conditions:

FP3 Pins dipped into 200 ul 0.6 mg/ml FITC in 100% DMSO.

Pins removed from source plate at 15 RPM (slow).

Pins touched off on nylon membrane or not touched off.

Pins transferred to 200 ul 0.1 M Tris, pH 8.0 in black polystyrene assay plate.

Hanging drop calculated by subtraction.

0.787 mm Diameter Pin (Uncoated FP3) Volume Transferred by Total Pin or Hanging Drop
Description	nl Transferred	CV%
Total Pin	72.66	9.8%
Hanging Drop	32.89	7.5%

Effect of Dwell Time on Dry Plate Transfers

Assay conditions:

FP3 Pin dipped into 200 ul 0.6 mg/ml FITC in 100% DMSO.

Pins removed from source plate at 15 RPM (.78 cm/sec).

Pins touched off in dry wells of black polystyrene assay plate**.

200 ul 0.1 M Tris, pH 8.0 was added to each well.

0.787 mm Diameter Pin (Uncoated FP3) Volume Transferred by Pin Tip
Dwell Time (Sec.)	nl Transferred	CV%
1	8.36	10.7%
2	8.43	11.1%
4	8.61	9.3%
10	9.36	8.9%

Detailed Assay Method:

Pin cleaning method