WPI PicoPump PV830 User manual

PV830
Pneumatic PicoPump
World Precision Instruments
INSTRUCTION MANUAL
Serial No._____________________
042712
www.wpiinc.com

PV830 Pneumatic PicoPump
World Precision Instruments iii
Copyright © 2012 by World Precision Instruments, Inc. All rights reserved. No part of this publication
may be reproduced or translated into any language, in any form, without prior written permission of
World Precision Instruments, Inc.
CONTENTS
ABOUT THIS MANUAL ..................................................................................................................... 1
INTRODUCTION.................................................................................................................................. 1
General ............................................................................................................................................. 1
Notes and Warnings .................................................................................................................... 1
Unpacking ....................................................................................................................................... 2
INSTRUMENT DESCRIPTION.......................................................................................................... 3
Setup.................................................................................................................................................. 5
Vacuum Input................................................................................................................................. 6
Pressure Input................................................................................................................................. 6
Eject Pressure Port (front panel)............................................................................................... 6
Vacuum Port ................................................................................................................................... 8
OPERATING INSTRUCTIONS........................................................................................................... 9
Front Panel ...................................................................................................................................... 9
Rear Panel .....................................................................................................................................10
Techniques In Microinjection ..................................................................................................10
Setting the Hold Pressure ...................................................................................................11
Micropipette Manufacture..................................................................................................12
Volume Calibration...............................................................................................................12
Multibarrel Microinjection..................................................................................................14
MAINTENANCE..................................................................................................................................14
Cleaning .........................................................................................................................................14
ACCESSORIES.....................................................................................................................................14
Replacement Parts......................................................................................................................15
TROUBLESHOOTING.......................................................................................................................15
SPECIFICATIONS................................................................................................................................16
Pressure..........................................................................................................................................16
Vacuum...........................................................................................................................................16
Timing.............................................................................................................................................16
Physical Specifications...............................................................................................................16
BIBLIOGRAPHY .................................................................................................................................17
DECLARATION OF CONFORMITY...............................................................................................18
WARRANTY ........................................................................................................................................19
Claims and Returns ....................................................................................................................19
Repairs ............................................................................................................................................19

iv World Precision Instruments

PV830 Pneumatic PicoPump
World Precision Instruments 1
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. 1—PV830 Pneumatic PicoPump
INTRODUCTION
The PV830 is designed to inject very small quantities of fluids, such as drugs,
into cells or small organelles. Pressure injection is an especially useful alternative
to electroionophoresis, since it does not mandate the use of charged ions. Two
different positive pressures may be applied–one for ejection at high pressure and
a second, lower pressure to prevent back filling of the pipette by capillary action
or diffusion. Vacuum may also be applied to hold cells or small objects and to load
pipettes from the tip. Cells may be held by vacuum and simultaneously injected
using pressure.
Notes and Warnings
!WARNING: WHEN USING A DELIVERY MICROPIPETTE, SECURE THE
PIPETTE FIRMLY. WHEN HIGH PRESSURE IS APPLIED, A LOOSE PIPETTE
CAN BE EJECTED FORCEFULLY. DO NOT APPLY PRESSURES IN EXCESS
OF 150PSI (1000KPA). USE DRY AIR, NITROGEN OR OTHER INERT GASES ONLY.
!WARNING: THIS INSTRUMENT IS FOR INVESTIGATIONAL USE ONLY IN
ANIMALS OR OTHER TESTS THAT DO NOT INVOLVE HUMAN SUBJECTS.

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Unpacking
Upon receipt of this instrument, make a thorough inspection of the contents and
check for possible damage. Missing cartons or obvious damage to cartons should be
noted on the delivery receipt before signing. Concealed damage should be reported
at once to the carrier and an inspection requested. Please read the section entitled
“Claims and Returns” on page 23 of this manual. Please contact WPI Customer
Service if any parts are missing at 941.371.1003 or [e-mail].
Returns: Do not return any goods to WPI without obtaining prior approval (RMA
# required) and instructions from WPI’s Returns Department. Goods returned
(unauthorized) by collect freight may be refused. If a return shipment is necessary,
use the original container, if possible. If the original container is not available, use a
suitable substitute that is rigid and of adequate size. Wrap the instrument in paper
or plastic surrounded with at least 100mm (four inches) of shock absorbing material.
For further details, please read the section entitled “Claims and Returns” on page
23 of this manual.

PV830 Pneumatic PicoPump
World Precision Instruments 3
INSTRUMENT DESCRIPTION
NO NC
Pressure Input
Filter
Pressure
Regulator
Pressure
Gauge
Pressure Gauge
Pressure
Regulator
Solenoid
Toggle Valve
(vac/hold)
Pressure Output
Vacuum Gauge
Vacuum
Regulator Filter
Vacuum Input
Filter
Inactive
Active
Hold
Vacuum
Toggle Valve
(Vac/Atm)
Atmosphere
Vacuum/Atm
Output
Fig. 2—The pnematic block diagram of the PicoPump is shown.
Both pressure lines (Hold and Eject) and the vacuum line pass through precision
regulators and are monitored by measuring gauges (Fig. 2). A fast solenoid
valve controls the application of the Eject pressure to the Pressure Port. When
the solenoid is inactive, the Pressure Port is connected to the Hold pressure.
Alternatively, the solenoid can be vented to the vacuum/atmosphere line, depending
upon the position of the Vac/Hold toggle switch. This feature allows the pressure
ejection channel to have a mild sucking action when pressure is not being applied.
A manual pneumatic switch also allows the vacuum/atmosphere line to be used
independently of the solenoid-controlled Pressure line.
Quantitative control over the amount of fluid injected is attained by adjusting the
pressure and the duration of the pressure pulse. For dispensing aliquots of as little
as tens of picoliters through micron-sized pipette tips, pressures of 10 to 100PSI are
used in pulses as brief as 10ms. Longer pulses, higher pressures and larger pipette
tip diameters give correspondingly greater amounts of fluid. For convenience, the
pressure is usually kept constant throughout a series of experiments and the pulse
width is varied to give different amounts of injected fluid.
The Startup Kit (WPI #3316) includes:
(1) 0.25” NPT Fitting for nitrogen tank regulators
(2) 0.25” quick connect fit barbs (use on vacuum side, where applicable)
(1) 5’ hard tubing

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Fig. 3—(Left) The PV830 Startup kit includes tubing and several fittings.
Fig. 4—(Right) The threaded fitting is for a nitrogen regulator. Depress the plastic collar to
remove tubing.
Fig. 5—The PicoPump Pressure and Vacuum ports are located on the rear panel of the
PV830.
To install the hard tubing, push it into the port until the tubing engages and cannot
be pulled out. To remove tubing, press in on the plastic collar. Do not remove tubing
holders. These parts are intended to remain on the pump.

PV830 Pneumatic PicoPump
World Precision Instruments 5
Setup
Firmly insert tubing into the quick-connect fittings on the rear of the instrument. To
remove tubing, press in on the surrounding collar while pulling the tubing out. Plastic
tubing can be disconnected easily, while metal tubing may require more effort. For a
diagram of the setup, see Fig. 6.
Air or
Nitrogen
Supply
150 psi max
Vacuum Pump or Line
Pressure InputVacuum Input
Hold Eject
Port
Micropipette Holder
Micropipette
Holder
Glass
Micropipette
Glass
Micropipette
Quick Connect Fitting
Collar1/4 in. Tubing
To insert, push the tubing in firmly.
To remove, push in on the collar
while pulling the tubing out.
Vacuum
Port
Fig. 6—This diagram show the experimental setup.

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Vacuum Input
Connect the Vacuum Input (located at the back of the instrument) to a suitable
source of vacuum, such as a vacuum pump (WPI #LM-500865) or aspirator, using
0.25˝ OD hard tubing and the soft plastic connector (supplied). Vacuum may be
anywhere in the range of 0–30 in. Hg. The vacuum line does not need to be
connected if only positive pressure is needed.
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 0–150PSI. Connect this
0.25˝ OD tubing to the quick connect fitting as indicated in Fig. 6.
A 0.25˝ FNPT fitting is supplied for connection to a nitrogen regulator on a N2
tank. Recommended gases are dry air, nitrogen or argon. Never use corrosive
gases. If an air pressure line containing oil or water vapor is used, an external filter
is recommended to prevent excessive contamination of the internal pneumatic
components.
!WARNING: THE PRECISION REGULATORS USED IN THIS INSTRUMENT
CONTINUOUSLY VENT A SMALL AMOUNT OF SUPPLY PRESSURE AS A
PART OF THEIR NORMAL FUNCTION. TO PREVENT WASTE OF GAS,
ALWAYS TURN OFF THE MAIN SUPPLY PRESSURE WHEN THE PV830 IS NOT IN
USE.
Eject Pressure Port (front panel)
Each PicoPump is supplied with two PicoNozzle Kits plus tubing to connect the
holders to the pressure and vacuum ports. Use one kit for pressure, one for vacuum.
Version 1 Kit (Optional)
Fig. 7—PicoNozzle Version 1 Kit (#5430-xx) allows micropipettes to be securely mounted
in micropositioners for stable axial air delivery. Because air enters the pipette axially,
lateral whipping during injection is eliminated.

PV830 Pneumatic PicoPump
World Precision Instruments 7
This kit 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, 0.120” OD, male locking Luer fitting on one end and a
female locking Luer fitting at the other end, rated for 200 PSI and 86 durometer
shore A)
The microelectrode holder is equipped with a female luer to attach to one end of the
4˝ male/male luer lock adapter. The 5´ tubing also has a female luer at one end to
attach to the opposite end of the 4˝ luer lock adapter. The male luer fitting of the 5´
tubing must be cut off in order to attach it to the pressure port of the PicoPump.
When using a MPH6S handle, a firmer hold on the glass can be achieved by using
two gaskets in the micropipette holder.
NOTE: Capillary holders can be obtained from WPI which contain Ag/AgCl half-cells.
These holders (MPH6S, MPH6R) can be easily mounted upon amplifier headstage
probes so that potential and/or current, as well as pressure, can be measured or
dispensed through the capillary tip.
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.
Version 2 Kit (Included)
Fig. 8—Each PicoPump is supplied with a 5430-ALL (Version 2) kit.
This kit contains:
(2) PicoNozzle tip assemblies (Handle diameter is 6.25 x 100mm.)

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(2) 5’ tubing
(4) 1.0mm pipette gaskets (green)
(4) 1.2mm pipette gaskets (black)
(4) 1.5mm pipette gaskets (red)
(4) 1.65mm pipette gaskets (white)
Handle Tubing
Tubing connected
to the barb
Body Gasket Cap
Pipette
Fig. 9—The 5430-ALL (version 2) must be properly assembled.
Before use, the superthane tubing must be slid through the handle and connected to
the barb of the body. Be careful so that the barb does not break when attaching or
removing the tubing. Before inserting the pipette, place a gasket of the correct size in
the cap.
Gasket Color Green Black Red White
Pipette Diameter (mm) 1.0 1.2 1.5 1.65
!WARNING: DANGER OF INJURY EXISTS IF THE PIPETTE IS INSECURE.
HIGH PRESSURE CAN CAUSE EJECTION AT HIGH VELOCITY.
Long lengths of tubing will 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) to the smallest possible value.
Vacuum Port
If this channel is to be used, connect the other PicoNozzle Kit to it exactly as
described above.

PV830 Pneumatic PicoPump
World Precision Instruments 9
OPERATING INSTRUCTIONS
Front Panel
Fig. 10—PV830 Front Panel.
!CAUTION: The valves always pass some air. Do NOT apply too much
pressure when turning the knobs. They will not reach zero while the PV830 is
running. Over-tightening the knobs will damage the regulators.
Pressure Adjustment–Adjust the pressure by turning the Regulator knobs located
next to the pressure gauges. Turn the knob clockwise to increase the pressure, and
counter-clockwise to decrease it. Because the regulating mechanism is self-venting,
pressure is automatically released when the regulator setting is decreased. Although
the pressure gauges cannot be read with high accuracy (especially at pressure
settings below 2PSI), very reproducible pressures may be obtained, because the
pressure regulators have a 20-turn dial yielding good pressure resolution.
The pressure regulating mechanism functions by continuously bleeding a small
amount of gas. This bleed rate increases as the difference between the input
pressure and the output pressure increases. Loss of gas may be minimized by
decreasing the difference between the input and output pressures, but some
decrease in regulation may be noticed if the input pressure is not at least 10%
greater than the output pressure.
!CAUTION: To conserve tank contents, make sure to turn off the gas supply
pressure when the PV830 is not in use.
Vent–The vac/hold switch selects between venting the Eject pressure port to either
the Hold pressure (hold) or to the vac/atm line (vac). If the vacuum line is not in use
and the switch is set to vac, the pressure port is vented to the atmosphere.
Vacuum Adjustment–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 counter-clockwise decreases the vacuum. Because

10 World Precision Instruments
the regulating mechanism is self- venting, vacuum is automatically released when
the regulator setting is decreased. Although the vacuum gauge cannot be read
accurately at very low vacuum settings (below 2 in. Hg), reproducible settings may
still be obtained by means of the 20-turn dial regulator knob.
Vacuum–When the Vacuum switch is in the vac position, the vacuum output port is
connected to the regulated vacuum line. When the switch is in the atm position, the
output port is vented to the atmosphere.
Power–Switching the Power on allows the solenoid of the Eject Pressure line to be
activated. An amber light above the switch indicates when power is on.
Timing–Electronic timing of the Eject pressure solenoid is effected by the control
section on the right hand side of the instrument front panel. Operation of the Eject
pressure solenoid can be initiated by means of any of three different methods:
• by manually pushing the Start button.
• by applying a +5V level at the EXT Input.
• by depressing the optional Remote footswitch. (The Remote connector is
used for accessing the footswitch only).
The length of the pulse is determined by setting the Duration switch. In the gated
mode the pressure solenoid opens and remains open as long as any of the three
commands is maintained. In the timed mode, an electronic timer controls the time
duration that the pressure solenoid stays open and the three commands serve only
to start the timer. Long pulses in the timed mode may be aborted by pressing the
Stop button. A green lamp next to the Eject pressure gauge illuminates for as long
as the pressure solenoid is open (energized).
Period and Range–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 1–101ms (every turn of the dial
is 10ms) and in the 10s range pulses may be set from 100ms to 10.1s (every turn of
the dial is 1.0s). The minimum time interval is limited by the speed of the solenoid,
which varies from approximately 10ms at 0 psi to 3ms at 100 psi.
Monitor–The Monitor output connector produces a logic-level output (+5 V)
corresponding to the time interval during which the Eject pressure solenoid is
energized. At all other times the monitor output is low (0V).
Rear Panel
A polarized, 3-conductor, connector is used for line (mains) power input to the
instrument. A removable cordset, terminated with a grounded 3-prong connector,
is standard. An alternate cordset may be supplied when local circumstances dictate
different mains voltages and connections.
A fuseholder contains a protective fuse in series with the high side (brown or black
wire) of the mains. The holder accepts 1/4by 11/4inch (6.35mm by 31.8mm) fuses of
the type indicated on the rear panel (120V, 0.5A–#6402 or 230V, 0.25A–#6409).

PV830 Pneumatic PicoPump
World Precision Instruments 11
Techniques In Microinjection
One of the more demanding tasks for which the PicoPump has been designed is the
microinjection of fluid into cells. Difficulties encountered will not usually involve the
mechanics of the PicoPump, but rather the manufacture of suitable micropipettes.
Both care and steady hands are needed in making pipettes to take full advantage of
the features of the PicoPump. Here we list several important things to keep in mind
when working with the PicoPump.
Setting the Hold Pressure
The Hold pressure is used to counterbalance the capillary action of the fluid
backfilling into the pipettes. When an empty pipette is inserted into the fluid, a
meniscus can be seen to rise up from the capillary tip. Hold pressure may be set
by adjusting the regulator setting (with the Hold/vent switch in the hold position)
until the meniscus stops at the desired position. In many applications, colored dye
or fluorescent dye is dissolved in the injection fluid. The capillary effect can be seen
with the color change at the tip of the pipette. When the fluid flows into the pipette,
the color of the tip becomes lighter. If the hold pressure is higher than the capillary
pressure, the fluid oozes out of the pipette. The solution around the pipette will be
colored. Adjusting the Hold pressure prevents this from happening.
Understanding how capillary action causes the backfilling of the pipette helps you
to correctly use the Hold pressure. The backfilling flow rate is determined by the
pressure of capillary action and the tip size. Since the tip size is often determined
by the requirement of the application, controlling the pressure of capillary action
becomes the main option to eliminate backfilling. 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, a
common misunderstanding.
If we assume the pipette tip is near a cylindrical shape, the pressure of capillary
action can be described by the LaPlace equation:
P=4γcosθ/d
where γis the surface tension, θis the contact angle between the water and glass,
and d is the inner diameter of the capillary where the meniscus is located. In most
cases, we can assume the contact angle for glass and water is zero (unless the
glass surface is treated). From this equation we can see that the capillary pressure
can vary a thousand times when the meniscus is moved from a 0.5µm ID tip to
the 0.5mm shank. The pressure at 0.5µm tip is about 80 PSI (in aqueous solution)
while at the shank will be only 0.08 PSI. Using one regulator to counterbalance the
pressure in such a large dynamic range is not practical. The hold regulator in this
instrument is optimized to work in the 0.2–10 PSI range. 10 PSI can counterbalance
a meniscus at the section of tip where the inner diameter is 4µm. In practice, this is
the highest pressure ever needed. On the lower pressure end, it becomes difficult to
exactly counterbalance the capillary pressure when the meniscus is at the shank of

12 World Precision Instruments
the pipette. However, a 0.1–0.2 PSI pressure imbalance will not cause a significant
problem if the tip is small enough. The gravity of the fluid and the flow resistance
caused by friction from the glass wall will both help to stop the solution flow at this
pressure level. If the lowest pressure setting still can’t stop the fluid from oozing
out, try to switch off the hold pressure to see if the gravity and friction are sufficient
to counterbalance the backfill. In addition, resetting the regulator could allow the
regulator to perform much better in this range (see the “Troubleshooting The Hold
Pressure” on page 16).
The capillary action can also be reduced by adding a hydrophobic fluid (such as
silicone oil) behind the hydrophilic saline solution. It can be completely eliminated
by silanizing the shank of the pipette (silanization increases the θin the LaPlace
equation to 90º).
Micropipette Manufacture
The volume of fluid ejected is markedly dependent on the micropipette tip size.
When using micron-sized tips a reduction in tip-size of a few percent may give an
order of magnitude difference in the flow rate. With tip sizes less than 1µm, pressure
ejection becomes increasingly difficult and special steps must be taken.
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. With 1µm and smaller
tips, capillary action becomes prohibitively large, and the hydrophilic surface of
the glass greatly limits the flow of fluid through the tip. Silanization decreases the
surface tension and allows the fluid to flow smoothly through the tip. For similar
reasons, use of a capillary with an internal filament is contraindicated. Some of the
many papers on the art of silanization are listed in the bibliography.
When using the vacuum line to hold and manipulate individual cells a large tip
(about 10–20% of cell size) is recommended. To prevent damage to the cell this tip
should be fire-polished.
Volume Calibration
For ejected volumes greater than 1nL visual inspection using a microscope can
be an accurate gauge of volume. A single pulse deposits a drop of fluid on the tip
of the micropipette. The volume of this drop may be calculated by measuring the
radius of the drop and assuming the drop to be spherical. Fig. 11 may be helpful
in determining the volume for a given radius. See “Appendix” on page 19 for a
comparison of spherical and cubical volumes.

PV830 Pneumatic PicoPump
World Precision Instruments 13
1 pl
10 pl
100 pl
1 nl
10 nl
100 nl
1 µl
10 µm 100 µm 1 mm
Volume of a Spherical Droplet V =
πd3
6
Volume
Diameter
Fig. 11—(Left) The volume of a droplet increases as the diameter increases. See “Appendix”
on page 16.
Fig. 12—(Right) WPI’s MicroFil is helpful in backfilling glass pipette tips. Syringe filters
(available separately) help prevent clogged micropipette tips.
The following table is useful for converting between different units of volume.
Cubic Measure 1cm31mm3(100µm)3(10µm)31µm3
Volume 1mL 1µL 1nL 1pL (10-12L) 1fL (10-15L)
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. If it is desired to silanize only the outside of the pipette, the PV830
may be used to good advantage by applying air flow through the pipette during the
silanization.
Precise assays of ejected volume may also be obtained through various
radioisotopic methods. See the bibliography for further information.
A slight deflection of the micropipette tip may be noticed during the application
of the pressure pulse. This deflection may be eliminated by ensuring that the
micropipette is firmly seated in the holder and that the holder is firmly held by a
manipulator.

14 World Precision Instruments
When pumping electrolytes, some indication of flow may be obtained by monitoring
the resistance or the voltage of the micropipette. If, for example, the micropipette
is filled with 3M KCl and the sample fluid is 0.1M KCl, a significant decrease in
resistance of the micropipette occurs when pressure is applied. This happens
because the resistance measured is determined primarily by the conductivity of
the solution at and within the tip. As pressure is applied the 0.1M KCl surrounding
the tip is replaced by the more highly conductive 3M KCl, decreasing the measured
resistance. Likewise, if vacuum is applied, the resistance increases as the less
conductive 0.1M KCl fills the tip.
NOTE: The above technique will not work with exterior and interior solutions of the
same concentration. However, a voltage change can be measured as a function of
the pressure applied. Working with tip sizes of 1–2µm, we have seen a change in
measured voltage of approximately 1mV for every 10 PSI of applied pressure.
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 PV830 to drive up to six micropipette barrels independently.
MAINTENANCE
The PicoPump has been designed to yield reliable performance. However particular
laboratory conditions may require occasional replacement of the pressure and
vacuum filters. Should this become necessary, return the instrument to the factory
for filter replacement.
Cleaning
Do not use alcohol, aromatic hydrocarbons or chlorinated solvents for cleaning.
They may adversely react with plastic materials used to manufacture the instrument.
The exterior of this instrument may be cleaned periodically to remove dust,
grease and other contamination. There is no need to clean the inside. Use a soft
cloth dampened with a mild solution of detergent and water. Do not use abrasive
cleaners.

PV830 Pneumatic PicoPump
World Precision Instruments 15
ACCESSORIES
Part Number Description
3260 Foot Switch
2932 Rack Mount Kit, 31 2-in. high (PV800 & PV820)
2933 Rack Mount Kit, 51 4-in. high (PV830)
5430-10 PicoNozzle 1 Kit (MPH6S for 1.0 mm pipette & 5-ft tubing
assembly)
5430-12 PicoNozzle 1 Kit (MPH6S for 1.2 mm pipette & 5-ft tubing
assembly)
5430-15 PicoNozzle 1 Kit (MPH6S for 1.5 mm pipette & 5-ft tubing
assembly)
5430-20 PicoNozzle 1 Kit (MPH6S for 2.0 mm pipette & 5-ft tubing
assembly)
5430-ALL PicoNozzle 2 Kit
MPH6S Micropipette Holder (specify 1.0, 1.2, 1.5 or 2.0 mm)
MPH6R Micropipette Holder (specify 1.0, 1.2, 1.5 or 2.0 mm)
3316 Replacement Input Kit
LM-500865 Vacuum Pump
Replacement Parts
Part Number Description
75122-110 PicoNozzle gasket green 1.0mm, pkg. of 10
75122-210 PicoNozzle gasket black 1.2mm, pkg. of 10
75122-310 PicoNozzle gasket red 1.5mm, pkg. of 10
75122-410 PicoNozzle gasket white 1.65mm, pkg. of 10

16 World Precision Instruments
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 0.5 PSI, but not below 0.5 PSI, a simple reset will fix the problem.
The reason that the regulator can’t reduce the pressure below 0.5 PSI is because
the dampening O-ring inside the regulator is stuck. The O-ring is connected to
the regulator’s diaphragm. It prevents the diaphragm from oscillating in high flow
conditions. The diaphragm moves the valve shaft. The diaphragm uses the output
pressure to counter-balance the spring force that is controlled by the knob. When
the output pressure becomes very low, the force generated on the diaphragm
becomes low. Sometimes, it is not enough to overcome the friction of the O-ring to
push the valve to the completely closed position. A high pressure from the output
port will push the diaphragm and return the O-ring to the seat.
To introduce a high pressure:
1. Remove the 1/4” hard pressure tubing from the back of the pump.
2. Use pieces of tubing with 1/4” OD to make a small 1/16” ID drop down
extension.
3. Position the extension inside the end of the 1/4” hard pressure tubing.
4. Place the other end of the extension on the Eject port
5. Then, apply an injection at 20 PSI for 10 seconds.
6. Once the O-ring re-seats, it will remain there even when the output pressure
again increases, as long as there is no high flow output. When there is no
restriction at the output and the pressure is adjusted too high, the diaphragm
has to move a substantial distance to allow the valve to open more to
compensate the for pressure loss. This also causes the O-ring to move.
Therefore, if low-pressure regulation is very important for the application, you
should not set the Hold regulator at the high-pressure position when there is no
restriction at the output. There are two ways to achieve this:
• Always reduce the regulator pressure to minimum before assembly and
disassembly of the injection pipette.
• Turn off the Hold pressure valve before assembly and disassembly of the
injection pipette.
NOTE: If you have a problem/issue with that falls outside the definitions of this
troubleshooting section, contact the WPI Technical Support team at 941.371.1003 or
[e-mail].