PV820/PV830 www.wpiinc.com Pneumatic PicoPump INSTRUCTION MANUAL Serial No.
Other WPI Favorites M3301 Manual Micromanipulator The world’s most widely used micromanipulator Weighing just 550 grams and employing a slim space-saving design, this well-built micromanipulator outsells all others worldwide for high precision experiments where magnification is in the range of up to 250. Its design allows units to stand tightly grouped — since all control knobs project to the rear.
PV820/PV830 Pneumatic PicoPump CONTENTS ABOUT THIS MANUAL ........................................................................................................................ 1 INTRODUCTION.................................................................................................................................... 1 PV820 ............................................................................................................................................... 2 PV830 .........................
iv World Precision Instruments
PV820/PV830 Pneumatic PicoPump ABOUT THIS MANUAL The following symbols are used in this guide: This symbol indicates a CAUTION. Cautions warn against actions that can cause damage to equipment. Please read these carefully. This symbol indicates a WARNING. Warnings alert you to actions that can cause personal injury or pose a physical threat. Please read these carefully. NOTES and TIPS contain helpful information. Fig.
amount of time that the injection pressure is applied to the output port. Time intervals can range from 10 seconds down to 10ms or less, depending on the eject pressure setting. The injection pressure interval can be triggered manually on the front panel, by foot switch or by a computer controlled TTL pulse. A 5V monitor output provides a logic-level pulse for your computer or other monitoring device.
PV820/PV830 Pneumatic PicoPump Parts List After unpacking, verify that there is no visible damage to the instrument. Verify that all items are included: (1) PicoPump (PV820 or PV830) (1) 3316 Startup Kit, which includes: • (1) 0.25” female NPT fitting for nitrogen tank regulators (Fig. 3.) • (1) Suction connector, silicone (Fig. 4.) • (1) 5’ length of hard pressure tubing, 1/4” OD. (Cut it in half to make pressure and vacuum lines.) (1) 5430-ALL PicoNozzle Kit (Fig.
Fig. 5—Each PicoPump is supplied with a 5430-ALL PicoNozzle (Version 2) kit. Legacy PicoNozzle Kits NOTE: The legacy Version 1 PicoNozzle Kits are still available and sold separately and also work well with the PicoPumps. Fig. 6—PicoNozzle Version 1 Kits (#5430-10, 5430-12, 5430-15, 5430-20). Version 1 PicoNozzle kits each contains: (1) MPH6S microelectrode holder (1) Handle for the MPH6S (4” hollow tube with male Luer fitting at both ends-handle diameter is 6.25 x 100mm.) (1) 5’ tubing (0.060” ID, 0.
PV820/PV830 Pneumatic PicoPump To mount the micropipette, pulled capillary glass may be inserted in the holder. A screw cap allows the glass micropipette to be firmly held by a rubber gasket. The luer fittings make changing micropipettes easy by allowing quick removal of the pipette holder from the 4˝ luer lock adapter. Test to make sure the micropipette is firmly held by pulling on it. Unpacking Upon receipt of this instrument, make a thorough inspection of the contents and check for possible damage.
INSTRUMENT DESCRIPTION PV820 Hold Pressure Gauge Eject Pressure Gauge Solenoid Controls PV830 Vacuum Pressure Gauge Secondary Vent Port rt Selector Switch Hold Pressure Gauge Secondary Vent Port SSeconda (Vacuum/Atmosphere) (Vacuum Eject Pressure essure e ge Gauge Solenoid SSo ole enoid Inputt Selector Switch Selector l h So Solenoid C Controls E Eject Port Power err Switch h Fig. 7—The major features of the PicoPumps are labeled in this diagram.
PV820/PV830 Pneumatic PicoPump ! CAUTION: The regulators leak a small amount of gas as a part of their normal operation. To conserve tank contents, make sure to turn off the main gas supply pressure when the PicoPump is not in use. Vacuum is adjusted by turning the vacuum Regulator knob located next to the vacuum gauge. Turning the knob clockwise increases the vacuum (lower pressure), and counterclockwise decreases the vacuum.
Typical Setups Most of the time, you will use one of two setups on the PicoPump: 2nd Vent Port Selector Switch* Solenoid Input Selector Switch** Result Hold a cell or reverse fill a pipette Vac Vent (PV820) Vac (PV830) Vacuum applied at the Eject port until solenoid is engaged Normal operations Vac Hold • Vacuum available at the Secondary Vent Port Purpose • Positive hold pressure applied at the Eject port until solenoid is engaged.
PV820/PV830 Pneumatic PicoPump Range Switch/Period Dial–The duration of the solenoid open time, in the timed mode, is determined by the 10-turn Period dial and the setting of the Range switch. In the 100ms range, the solenoid times may be set from 0.1–100ms (every turn of the dial is 10ms). In the 10s range, pulses may be set from 10ms to 10s (every turn of the dial is 1.0s).
Installing Hard Tubing 1/4 in. Tubing Collar Quick Connect Fitting To insert, push the tubing in firmly. To remove, push in on the collar while pulling the tubing out. Fig. 9—The PicoPump Pressure and Vacuum ports are located on the rear panel. ! CAUTION: Do not remove tubing holders. These parts are intended to remain on the pump. To install the hard tubing, firmly insert tubing into the quick-connect fittings on the rear of the instrument.
PV820/PV830 Pneumatic PicoPump Pressure Input–Connect the Pressure Input (located at the back of the instrument) to a suitable source of pressure, such as a compressed gas tank or an air line, using 0.25˝ OD tubing supplied. Pressure may be anywhere in the range of 0–150 PSI. Connect this 0.25˝ OD tubing to the quick connect fitting. A 0.25˝ female NPT fitting is supplied for connecting to a nitrogen regulator on a N2 tank. Recommended gases are dry air, nitrogen or argon.
Gasket Color Pipette Diameter (mm) 3. Green 1.0 Black 1.2 Red 1.5 White 1.65 Connect the other end of the small tubing to the appropriate port on the front of the PicoPump. ! WARNING: DANGER OF INJURY EXISTS IF THE PIPETTE IS INSECURE. HIGH PRESSURE CAN CAUSE EJECTION AT HIGH VELOCITY. NOTE: Long lengths of tubing tend to decrease the time response of the system due to a loading effect of the increased volume. For fastest possible response, keep the tubing length (and diameter) as small as possible.
PV820/PV830 Pneumatic PicoPump NOTE: The pressure of capillary action is determined by the inner diameter of the glass capillary where the meniscus of air/liquid interface is located. It has nothing to do with the pipette tip size.
The most important of these steps is cleaning the glass. Small amounts of dust or grease can easily clog micron-sized tips. Cleaning with chromic acid solutions before pulling the electrode is commonly performed, but care must be taken to thoroughly rinse the pipettes to remove all traces of the chromic acid, which has some affinity for glass. Some researchers prefer hydrochloric or nitric acid. Silanization of the glass is also recommended for small tips.
PV820/PV830 Pneumatic PicoPump For ejected volumes less than 1nL, visual inspection in air proves to be difficult due to rapid evaporation. The same technique may be used though if the drop is kept submerged under oil. Droplets may seem to disappear after emergence from the tip. Sometimes this is due to creepage of the aqueous fluid back along the outside shank of the micropipette. This creepage may be decreased by silanizing the outside of the pipette.
Solenoid Input Selection Switch. When the pipette is full, set the Solenoid Input Selection Switch to Hold. 7. Set the Duration switch to Gated. Hold down the foot switch to eject fluid from the micropipette into the oil-filled dish. 8. Record the amount of time it takes in seconds for the meniscus in the micropipette to travel from one 1mm mark to the next. It should take between 20 and 30 seconds for each injection. You may need to adjust the eject pressure to achieve this time frame. 9.
PV820/PV830 Pneumatic PicoPump Multibarrel Microinjection For injection with multibarrel micropipette, the PolyFil multibarrel micropipette coupling kit can be purchased from World Precision Instruments. This multibarrel micropipette coupling kit allows easy and secure coupling of a multi-barrel micropipette to a pressure source. Kits include a five-port manifold which allows use of a single PicoPump to drive up to six micropipette barrels independently.
ACCESSORIES Part Number 3260 2932 2933 5430-10 5430-12 5430-15 5430-20 5430-ALL 5440 MPH6S MPH6R 3316 LM-500865 Description Foot Switch Rack Mount Kit, 31 2-in. high (PV800 & PV820) Rack Mount Kit, 51 4-in. high (PV830) PicoNozzle 1 Kit (MPH6S for 1.0 mm pipette & 5-ft tubing assembly) PicoNozzle 1 Kit (MPH6S for 1.2 mm pipette & 5-ft tubing assembly) PicoNozzle 1 Kit (MPH6S for 1.5 mm pipette & 5-ft tubing assembly) PicoNozzle 1 Kit (MPH6S for 2.
PV820/PV830 Pneumatic PicoPump TROUBLESHOOTING THE HOLD PRESSURE If the Hold regulator can’t reduce the pressure low enough to keep the liquid from coming out of the pipette tip, DO NOT try to shut off the pressure by forcing the regulator knob counterclockwise. The regulator is designed so that the flow can’t be shut off completely. It should, however, be able to reduce the pressure to below 0.2 PSI, which is low enough for most applications. If the regulator can control the pressure above 0.
SPECIFICATIONS This unit conforms to the following specifications: PV820 PV830 PRESSURE Pressure Input 0 to 150 psi 0 to 150 psi 0.3 to 90 psi * 0.3 to 90 psi Pressure Output Pulse Width (10-Turn Dial) 10 ms to 10 s in Timed Mode 10 ms to 10 s in Timed Mode Regulator Accuracy 0.1% (20-turn dial) * 0.1% (20-turn dial) * 0.05 psi * 0.05 psi * Regulator Repeatability Gauge Accuracy 3% at full scale * 3% at full scale * Input Connector Quick Connect (1⁄4 in. OD Tubing) Quick Connect (1⁄4 in.
PV820/PV830 Pneumatic PicoPump APPENDIX A: FLOW DIAGRAMS The following diagrams depict how air and vacuum are routed through the PV820 and PV830 pumps. Air Source PV820 HOLD PRESSURE EJECT PRESSURE pressure Vacuum Source hold vac atmosphere Vaccum Port Output (vacuum or air) Solenoid Trigger/ Timing vent Eject Port Output (output of the solenoid = vacuum, eject pressure or hold pressure) Fig.
Schematics PV820 Block Diagram Filter Pressure Input Eject Pressure Output Port Solenoid Eject Pressure Gauge Pressure Regulator NO Active NC Inactive Pressure Regulator Toggle Valve (hold/vent) Hold Hold Pressure Gauge Filter Filter Hold Pressure Output Port Toggle Valve (Pressure/Vac) Vacuum Input Vacuum Atmosphere Fig. 17—The schematic diagram shows the inner workings of the PV820 PicoPump.
PV820/PV830 Pneumatic PicoPump APPENDIX B: DROPLET VOLUME Use the chart below to gauge the volume of a droplet and the table to determine the volume a micropipette can hold. 1cm 215μm 1mL = 1.24cm 10nL = 4.64mm 267μm 100μm 100μL = 5.76mm 1nL = 2.67mm 100pL = 1.24mm 10pL = 2.15mm 1.24μm 46μm 10μL = 1mm 57μm 21.5μm 1μL = 464μm 27μm 10μm 100nL = 576.36μm 1pL = 12.4μm Fig. 19—This graph compares the dimensions of a cube and a sphere with the same internal volume.
BIBLIOGRAPHY R.E. McCaman, D.G. McKenna and J.K. Ono, A Pressure System for Intracellular and Extracellular Ejections of Picoliter Volumes , Brain Research 136 141-147 (1977) T.W. Stone, Microintophoresis and Pressure Ejection (1985) Y. K llstrom and S. Lindstr m, A Simple Device for Pressure Injections of Horseradish Peroxidase into Small Central Neurons , Brain Research 156 102-105 (1978) R.E. Hammer, et al. Production of Transgenic Rabbits, Sheep and Pigs by Microinjection , Nature 315 680-683 (1985) R.
PV820/PV830 Pneumatic PicoPump INDEX Symbols 20-turn regulator 1 3316 3 5430-ALL 3 A argon 11 atmospheric air 7 C capillary action 12 chromic acid 14 cleaning 17 cordset 9 E eject port 7 electroionophoresis 2 external pulse generator 9 External Trigger Input Connector 8 F foot switch 2, 7, 8, 9, 16 fuse 9 G gasket 11 gaskets 3 Gated 9 gauges 1, 6 H Hold pressure 12 hydrochloric 14 L lamp 8 light 7 M Monitor 9 multibarrel micropipette 17 World Precision Instruments N nitric acid 14 nitrogen 11 P parts lis
DECLARATION OF CONFORMITY 26 World Precision Instruments
PV820/PV830 Pneumatic PicoPump WARRANTY WPI (World Precision Instruments, Inc.) warrants to the original purchaser that this equipment, including its components and parts, shall be free from defects in material and workmanship for a period of one year* from the date of receipt. WPI’s obligation under this warranty shall be limited to repair or replacement, at WPI’s option, of the equipment or defective components or parts upon receipt thereof f.o.b. WPI, Sarasota, Florida U.S.A.
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