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

     
 

 

 
 

 
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%
Hydrophobic 7.46 5.4%
0.229 mm diameter
(FP9)
Hanging Drop Uncoated N/A N/A
Hydrophobic 2.09 3.8%
0.457 mm diameter (FP1) Total Pin Uncoated 33.48 3.2%
Hydrophobic 28.17 7.5%
0.457 mm diameter
(FP1)
Hanging Drop Uncoated 16.96 4.5%
Hydrophobic 8.51 0.8%
0.787 mm diameter (FP3) Total Pin Uncoated 87.32 3.9%
Hydrophobic 77.40 3.9%

0.787 mm diameter
(FP3)

Hanging Drop Uncoated 48.77 1.2%
Hydrophobic 43.05 9.4%
1.19 mm diameter
 VP 409 & VP 386 
Total Pin Uncoated 247.22 2.8%
Hydrophobic 192.67 2.6%
1.19 mm diameter
 VP 409 & VP 386
Hanging Drop Uncoated 76.35 1.6%
Hydrophobic 108.40 2.8%
1.58 mm diameter
 VP 408 & VP 384
Total Pin Uncoated 273.50 4.6%
Hydrophobic 259.25 3.1%
1.58 mm diameter 
 VP 408 & VP 384
Hanging Drop Uncoated 201.93 5.0%
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%
Hydrophobic 3.61 6.8%
0.229 mm diameter (FP9)  

Hanging Drop**

Uncoated 0.90 5.6%
Hydrophobic 1.29 8.8%
0.457 mm diameter (FP1) Total Pin Uncoated 16.58 3.9%
Hydrophobic 16.06 2.9%
0.457 mm diameter (FP1) Hanging Drop Uncoated 3.74 1.9%
Hydrophobic 0.66 7.2%
0.787 mm diameter
(FP3)
Total Pin Uncoated 71.11 4.1%
Hydrophobic N/A N/A
0.787 mm diameter
(FP3)
Hanging Drop Uncoated 50.8 12.1
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%
Hydrophobic 170.20 2.9%
0.787 mm diameter (FP3)  

100 nl Slot 
Slot Only

Uncoated 149.67 4.9%
Hydrophobic 129.61 7.6%
0.787 mm diameter (FP3) 200 nl Slot Total Pin
Including Slot
Uncoated 269.77 1.9%
Hydrophobic 228.62 17.1%
0.787 mm diameter (FP3) 200 nl Slot
 Slot Only
Uncoated 237.52 8.9%
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%
Hydrophobic N/A N/A
10 nl Slot Total Pin* Uncoated 23.36 6.1%
Hydrophobic 25.85 6.9%
50 nl Slot Total Pin* Uncoated 67.83 2.5%
Hydrophobic N/A N/A
0.787 mm diameter (FP3)   100 nl Slot Total Pin*

Uncoated

180.32 7.2%
Hydrophobic 205.84 5.5%
200 nl Slot Total Pin* Uncoated 277.82 4.9%
Hydrophobic 287.30 3.8%
500 nl Slot Total Pin* Uncoated 581.16 5.2%
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

     
         

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