2pk+_manual

2pk+_manual
Model # 2PK+
INSTRUCTION MANUAL
Ver. August.2012
ALA Scientific Instruments Inc.
60 Marine Street
Farmingdale, NY 11735
Tel.
# 631.393.6401
FAX: # 631.393.6407
E-mail:
mail: [email protected]
www.alascience.com
Page 1 of 20
Table of Contents:
Page #
INTRODUCTION ............................................................................................................. 3
PARTS LIST .................................................................................................................... 4
COMPONENTS .............................................................................................................. 5
SET-UP ........................................................................................................................... 7
QUARTZ CAPILLARY PREPARATION .......................................................................... 9
WHOLE CELL PERFUSION SET UP ........................................................................... 10
Whole Cell perfusion Setup with unprepared Quartz Capillary ............................. 11
Whole Cell perfusion Setup with prepared Quartz Capillary ................................ 12
PERFUSION PATCH PIPETTES .................................................................................. 13
Patch perfusion Setup with unprepared Quartz Capillary ..................................... 14
Patch perfusion Setup with prepared Quartz Capillary ......................................... 14
HELPFUL HINTS .......................................................................................................... 15
LIMITED WARRANTY................................................................................................... 16
DIAGRAMS ................................................................................................................... 17
Block Diagram ...................................................................................................... 17
Valve Modes to be Avoided .................................................................................. 17
SPECIFICATIONS ........................................................................................................ 18
PRESSURE CONVERSION TABLE ............................................................................. 19
REFERENCE ARTICLES.............................................................................................. 20
Page 2 of 20
Introduction
The 2PK+ is designed to enable the perfusion of whole ceIIs, patch pipettes and sharp electrodes during
electrical recordings and/or imaging studies. The key element in the success of the technique is the
inclusion of a quartz microperfusion capillary. The quartz capillary is coated with polyamide and is
extremely flexible and durable. Its durability allows it to be positioned near the tip of the recording pipette.
Quartz has excellent low noise characteristics which helps to limit the amount of noise introduced during
perfusion. This insures that high quality recordings can be obtained even as perfusion is taking place.
Perfusion in the 2PK (Pipette Perfusion Kit), the predecessor to the 2PK+, was achieved by simply
increasing the amount of negative pressure applied to the patch pipette, This caused the perfusion
solution to be drawn into the patch pipette after the negative pressure reached a sufficient level to
overcome the restriction to flow caused by the ID of the quartz microcapillary. A precision vacuum control
unit was employed so that the flow could be achieved without destruction of the cell or the patch.
In order to be able to perfuse whole cells, increasing suction would cause the contents of the cell to be
pulled into the pipette possibly clogging the tip of the pipette. To overcome this obstacle, the 2PK+ uses
positive pressure and a pressure vessel that permits fluid to be injected into the recording pipette under
pressure. The cell can be protected by leaving the pipette open to atmosphere. Conversely, negative
pressure can be applied to neutralize the positive pressure so that the cell experiences no net change in
pressure.
The process of cell perfusion is achieved when the lumen of the recording pipette is open to the interior of
the cell. Fluids and other objects are free to pass between the cell and the pipette and visa versa. There
is a natural tendency for fluid in the pipette to mix into the cell. However, the 2PK+ assures the mixing by
setting up a stream of solution that flows from the perfusion capillary into in recording pipette and then out
into the cell. The increased volume of fluid can be displaced up the pipette. Since such a small volume
of fluid is transferred for a particular perfusion event, many perfusions are possible before the pipette is
backfilled to the rim. Also it can take as long as an hour for a previous solution to back diffuse down the
pipette (see Reference Articles).
With the 2PK+, it is possible to perfuse whole cells, patch pipettes and sharp electrodes by changing the
recording media in the pipette and the internal milieu of the cell. In some instances it may be as though
there is an IV directly into the cell whereby the researcher has continuous access to the interior of the cell
for the introduction of agonists, antagonists and fluorescent dyes.
Page 3 of 20
Parts List
Pressure/Vacuum Generator
120 230
9 Volt DC wall power supply
Perfusion Pipette Holder
Accessories kit
Pressure Vessel (1)
3 reservoirs 0.125ml
Tube and double luer valve assembly (1)
PE tube (2 pieces of PE-10, 1’ long)
Silver wire ( 3 pieces, 2” long)
Butane torch (1 with instructions)
Sand paper (1)
30 ga needle (4)
Vacuum grease (1)
Quartz tubing (20 pieces, 100 micron ID)
Beem capsules (10 large, 10 small)
Alligator clips (4)
Polystyrene block (1)
Input hose (2 pieces 1/8” id TYGON tubing)
Instruction Manual
Options
MRC-6 with 3 reservoirs
Page 4 of 20
Components
1)
Pressure / Vacuum Generator
Used to give precise control of pressure and vacuum for perfusion. It consists of two precise 20 turn
pressure regulators and an internal input regulator set to 15 PSI. An LCD display indicates the pressure
being monitored (in mmHg) as selected via the mode switch. Pressure and vacuum cannot be monitored
simultaneously. Pressure and vacuum readings can only be taken when the system is hooked in with the
other components. The vacuum side should not be left open to air when on. This may introduce
contaminants such as dust that will decrease performance. A five micron filter is incorporated in the
vacuum line to prevent damage to the system in event the pipette is damaged and fluid is sucked back.
Vacuum is created by a venturi pump (positive air flow creates vacuum). The vacuum pressure regulator
adjusts the air flow into the venturi which regulates the vacuum pressure. Both vacuum and pressure are
measured.
When the unit is in operation it will sound like there is an air leak as it always makes a hissing noise. The
higher the vacuum/pressure setting the louder the hiss. This is part of normal operation.
The vacuum pressure generator is designed to produce at least 55 mm/Hg of pressure and vacuum
respectively (The pressure side can go higher but it is not recommended for safe operation of the
generator). This can give an effective pressure difference to the perfusion solution of 100 mm/Hg. Enough
to make it fly into the pipette!
2)
Power Supply
This is a 9V DC wall adapter used to power the pressure/vacuum generator.
3)
Perfusion Pipette Holder
The perfusion pipette holder is specifically designed for this purpose. It is made of polycarbonate and is
available in different configurations to match any amplifier or pipette size (customer specified). Just like a
standard patch pipette holder it has a port for a suction tube. It also has an additional port where a PE
tube can enter and easily access the pipette lumen. The PE tube can be locked in to keep the holder
sealed.
4)
Positive Pressure Vessel
The positive pressure vessel is used for whole cell perfusion. It allows the user to push rather than suck
solutions into the pipette. For whole cell perfusion, adding suction to the pipette might cause the pipette
tip to clog. The pressure vessel has a port for pressure to enter and another one for the PE tube that
carries the perfusion solution to the pipette. The perfusion solution is sealed into the micro vial at its
bottom. Solutions can be changed simply by unscrewing the vial and screwing in another. The volume is
deliberately kept small since so little is required to perfuse a cell and since some substances are
considered precious. As an option, the MRC-6 can be used with the positive pressure vessel to provide
for multiple solution changes. See the MRC literature & manual for more information.
5)
Output Hose and Luer Valves
This is the required assembly to connect the perfusion pipette holder and the positive pressure vessel
together with the rest of the system. The luer valves are used to divert pressures toward and away from
components and also to open the system to atmosphere or connect to mouth or syringe suction.
6)
Silver Wire
It is used to provide an electrical connection from the pipette to the head stage of the amplifier. It should
be chlorided before use.
Page 5 of 20
7)
Polyethylene Tubing - (PE)
Thin polyethylene tubing is used for the linkage of the quartz capillary to the positive pressure vessel. PE
tubing provides an inert and low volume transfer pathway for perfusion solutions. Its narrow ID means
that dead volumes are kept to a minimum when solutions are changed.
8)
Sand Paper
Used for one particular technique for removing carbon debris from the quartz capillary after melting. It
can also be used to bevel the tip of the quartz perfusion capillaries.
9)
30 Gauge Needle
Thirty gauge needles are provided to enable the priming of the perfusion tube (PE tube) and the quartz
capillary. This assembly must be filled with the pipette filling solution before a seal is made. This
prevents air from entering the pipette and also prevents premature introduction of the perfusion solution.
10)
Vacuum Grease
It is used to enhance various seals on the pipette holder and positive pressure vessel if necessary.
11)
Quartz Tubing
Quartz tubing is used for the perfusion canula to carry the perfusion substance down through the pipette
to the tip. The quartz is provided in raw form and can be pulled and cut before use. It is coated with
polyamide and thus has a brown appearance. As long as the polyamide coating is present, the quartz is
extremely flexible.
12)
Beem Capsules
Used for holding solutions for patch pipette perfusion.
13)
Polystyrene Block
A Polystyrene block is used to hold the positive Pressure vessel or the Beem capsules during
experiments. It provides a convenient way to hold these small components near the perfusion pipette and
also provides valuable electrical insulation to further reduce noise.
14)
Additional Reservoirs
Three additional reservoirs are provided for the positive pressure vessel. They allow for quick change of
solution.
15)
Alligator Clips
These are used for pulling the quartz capillaries. One may be used to pin the quartz up while one, two, or
three are used to weigh down the other end.
16)
Butane Torch
The butane torch provides a small size flame of adequate temperature to melt the quartz tubing at around
0
700 C. The torch will run for about 15 minutes on a full charge. It can be recharged using regular butane
that is used in cigarette lighters. Torched is shipped empty for safety reasons.
Page 6 of 20
Set-up
Upon opening the kit, first check to be sure that all parts on the check list are included. If any part is
missing or damaged contact the supplier immediately.
The tubing and luer valve assembly must be connected to the front of the pressure/vacuum generator
(PVG). Connect the assembly as shown in the above diagram.
The Vacuum Line will get connected to the pipette holder side port. The Positive Pressure line will
connect to the Pressure Vessel when performing whole cell perfusion.
Page 7 of 20
The rear panel of the 2PK+ Pressure/Vacuum Generator has 2 connections.
1. Power In - Connect the 9V dc wall adaptor power supply to this port.
2. Input Pressure – Connect positive pressure either from a tank or house air. The pressure used
must be between the stated pressure ranges. The recommended pressure value is 25PSI.
Before air or gas is supplied, test the unit by turning on the power. With no pressure supplied the
pressure on the LCD should be zero or below +/- 1.0 mmHg. The power switch is a push-in and lock
type. Power should be turned off when the unit is not in use. It is also a good practice to turn off the air
supply to the unit when it is not required.
The vacuum line should not be left open while vacuum is on. This can cause dust and debris to enter the
vacuum mechanism and reduce performance.
Page 8 of 20
Quartz Capillary Preparation
The quartz capillary (supplied) necessary for perfusion can be used as is or can be made to fit further
down the pipette electrode. The quartz capillary supplied is 100um ID and 230um OD. If used as is, the
quartz capillary will not be able to get close to the cell as possible. To get close to the cell and speed up
the response rate the quartz capillary needs to be modified.
Below are instructions to prepare the quartz capillary to reach further down the pipette.
(Note: Many users find that the quartz capillary can be used without pulling the tip smaller. It is up to the
user to determine if this method will work. The smaller pulled tip can fit further down the pipette, but once
the polyimide is burned off the tip can break. The un-pulled tip is more robust, but will not fit as far down
the pipette, it will give faster flow, however, than the pulled one.)
Remove a length of quartz tubing from the vial
and suspend it vertically about eight inches
(20cm) above a clean work bench using one of
the alligator clips. Use one, two, or three other
alligator clips to weight down the other end.
Be sure that the capillary hangs straight and
steady.
Using the butane torch at a high flame, heat
the capillary at the midpoint until it melts and
the lower end falls. Move the flame away
quickly when this happens so as not to destroy
the upper strand. The polyimide will burn
away and reveal the quartz glass underneath.
The exposed quartz is very hard and brittle.
The carbon on the pulled part of the quartz must be
removed. This can be done with a small piece of
sandpaper or a micro knife.
A dissecting
microscope is very helpful. In addition, the capillary
can be cleaned with a sonicator and methanol.
Finally, the tip of the quartz is cut back using a pair
of scissors, wire cutters or other implements. The final tip size should be no less than 35µm. Larger
sizes can be experimented with. With the advent of the positive pressure vessel, it may be possible to
perfuse without pulling the quartz capillary. However the small size of the tip will help place it further down
into the pipette tip. It should not be necessary to make a fresh quartz capillary for each experiment. One
capillary should last through several experiments before requiring a change.
Page 9 of 20
Whole Cell Perfusion Set Up
For an experiment in which whole cell perfusion is to be performed, the 2PK+ must
be assembled as follows:
1.
2.
3.
4.
5.
6.
7.
8.
9.
10.
11.
12.
13.
14.
15.
16.
17.
18.
Connect the positive pressure line to the pressure vessel port.
Connect the vacuum line to the pipette holder barb port.
Connect the pipette holder to the headstage.
O
Place the PE-10 tubing down the 30 port on the holder until it is even with
the silver wire.
Place a quartz perfusion capillary made in accordance with the directions
above, into the PE tube adjacent to the silver wire.
Adjust the perfusion capillary by pulling and pushing the PE-10 tube at the
O
30 port until its tip is just ahead of the silver wire.
Using a thirty gauge needle and syringe, fill the PE tube/quartz capillary
with the pipette filling solution.
Place the recording electrode over the quartz capillary and silver/silver chloride wire and seal it
into place with the thumb screw.
O
Use the PE tubing at the 30 port to push the quartz capillary down to the tip of the pipette. The
tip of the capillary should sit at a point where its tip is roughly half the diameter of the patch
O
pipette. Tighten the 30 port to hold the perfusion capillary in position and to maintain a good
O
seal (add vacuum grease if necessary). The rubber gasket of the 30 port can be sealed on the
quartz tube, the PE tube or the interface of the two. Be sure it is very tight when sealing on the
quartz tube. Here vacuum grease should be employed for a good seal. In general, never let the
PE tube make contact with the solution in the recording pipette.
Prepare solutions for perfusion in the reservoirs for the positive pressure vessel. You should
begin with the pipette filling solutions. As experience is gained it may be possible to start with a
test solution as long as the PE tube and perfusion capillary are first filled with the pipette solution
just in case there is any pre-flow.
Seal the reservoir into the positive pressure vessel while the PE tube is still loose. This will
prevent the pressure produced when sealing from starting premature perfusion.
Insert the PE tubing down the top of the Pressure Vessel until it is at the bottom of the reservoir.
The PE tube connecting the pipette holder to the pressure vessel should be kept as short as
possible. This speeds up perfusion and helps reduce stray capacitance which is a source of
noise.
Set up the polystyrene block so that the positive pressure vessel can be placed in it and held
there during an experiment.
Set the pressure 3-way valve with the OFF position toward the vacuum side.
Set the vacuum 3-way valve with the OFF position toward the unused port. The unused port can
be used to form a seal using a syringe attached to it or by mouth suction.
The system is now ready to do a whole cell perfusion.
Form a seal to the cell as usual. Be sure that the positive pressure vessel is ported to vacuum as
in the diagrams to prevent premature perfusion when going on-cell.
Rupture the cell membrane using current (tweaking the electrode) or increased suction. A
syringe may be necessary for this, DO NOT PORT HIGH PRESSURE FROM A SYRINCE TO
THE 2PK+ PRES./VAC. GENERATOR AS IT MAY DAMAGE THE PRESSURE TRANSDUCER.
19. To perfuse whole cells, open the vacuum side of the pipette holder to atmosphere and increase
positive pressure on the pressure vessel (Be sure to use proper valve configuration --see
Diagrams). Positive pressure can also be offset using vacuum. Switch modes on the
Press./Vac. Generator back and forth to supply exact opposite pressures at first. Be advised that
the pressure that the perfusion solution experiences are equal to the absolute value of positive
and negative pressures applied to the system. Example: +10 mm/Hg applied to the positive
pressure vessel and 10mm/Hg applied to the suction port on the pipette holder gives the
perfusion solution an effective pressure force to enter the pipette of 20mm/Hg. Using the 2PK+ at
Page 10 of 20
its maximum, an effective pressure gradient of over 100mm/Hg can be achieved! In some cases
it may be necessary to balance pressures, in other cases simply opening the pipette to
atmosphere is enough. 10-20mm/Hg should be enough to start flow.
Please note: (1) With time, the user typically develops a feel for his/her particular set up. (2)
Rarely will positive and negative pressures be matched for perfusion to take place. This it is a
good starting point for first time users. The user may find that perfusion takes place with a few
mmHg of positive pressure and two or three times the amount of negative pressure or vice versa.
It must be discovered empirically since each experimental set up is different.
20. Be sure to neutralize all pressures after perfusion has taken place so as not to over fill the
recording pipette or introduce air bubbles. To neutralize, open the pipette holder suction and the
positive pressure vessel to atmosphere using the luer valves.
21. When changing reservoirs on the positive pressure vessel be sure that there is no suction being
applied. Also be sure the PE tube is loose and the positive pressure line is open to atmosphere
at the luer valve so that premature perfusion does not take place.
Those who need to maintain some suction on the pipette to hold the seal should take special care
to adjust the system to allow this without causing perfusion.
Whole Cell perfusion Setup with unprepared Quartz Capillary
Page 11 of 20
The above diagram shows the system used with the quartz tube in its original size (not prepared).The
quartz tube has a 100um id and will allow for higher flow rates when used in this manner. However, this
also impedes the quartz tube from getting close to the cell as the pipette narrows at the end. Being closer
to the cell increases the response rate.
The diagram below shows the system used with a prepared quartz tube. As you can see the quartz is
able to go further down the pipette when the quartz tube is pulled and a fine tip is formed.
Whole Cell perfusion Setup with prepared Quartz Capillary
Page 12 of 20
Perfusion Patch Pipettes
The set up for pipette perfusion is similar to the set up for whole cell perfusion
except it is a little simpler. The positive pressure vessel and the positive pressure
side of the 2PK+ does not need to be used, however it can be incorporated once
practice is gained in the technique.
For pipette perfusion, set up the perfusion pipette holder as below and lead the PE
tube to the polystyrene block where it will sit in a Beem capsule of solution, since
only suction is necessary for perfusion.
1. Prepare materials for perfusion in a small Beem capsule and place it in the
polystyrene block holder.
2. Connect the pipette holder to the headstage.
O
3. Place the PE-10 tubing down the 30 port on the holder until it is even with
the silver wire.
4. Place a quartz perfusion capillary made in accordance with the directions
above, into the PE tube adjacent to the silver wire.
O
5. Adjust the perfusion capillary by pulling and pushing the PE-10 tube at the 30 port until its tip is
just ahead of the silver wire.
6. Using a thirty gauge needle and syringe, fill the PE tube/quartz capillary with the pipette filling
solution.
7. Place the recording electrode over the quartz capillary and silver/silver chloride wire and seal it
into place with the thumb screw.
O
8. Use the PE tubing at the 30 port to push the quartz capillary down to the tip of the pipette. The
tip of the capillary should sit at a point where its tip is roughly half the diameter of the patch
O
pipette. Tighten the 30 port to hold the perfusion capillary in position and to maintain a good
O
seal (add vacuum grease if necessary). The rubber gasket of the 30 port can be sealed on the
quartz tube, the PE tube of the interface of the two. Be sure it is very tight when sealing on the
quartz tube. Here vacuum grease should be employed for a good seal. In general, never let the
PE tube make contact with the solution in the recording pipette.
9. Lower the pipette into the bath and place the end of the PE tube into the Beem capsule
containing the first solution to be perfused.
10. Patch the cell as normal. If you are using the positive pressure vessel, be sure to port suction to
it as well so that there will be no pre-flow of solution.
11. When ready for perfusion simply turn over suction control to the vacuum generator on the 2PK+.
Increase vacuum on the pipette holder. 20-30 mmHg should be enough to cause perfusion. Use
more if necessary. If using the positive pressure vessel, increase pressure to the positive
pressure vessel, but be sure to balance it off with at least 10 mmHg greater negative pressure to
maintain the patch. IT SHOULD BE NOTED THAT POSITIVE AND NEGATIVE PRESSURES
USED IN CONJUCTION WILL ALMOST NEVER BE EVEN. One may find that 10 mmHg
suction might be balanced by 40mmHg pressure or vice versa for example. These levels must be
determined empirically in each set up.
12. When perfusion is to be stopped, return the system to normal suction pressure for maintaining the
patch. If using the positive pressure vessel, port suction (negative pressure) back to it to prevent
additional filling of the pipette.
Perfusion solutions can be changed anytime perfusion is stopped. Depending on the rate of perfusion,
they might be able to be changed without introducing an air bubble. Perfusion capillaries should be
changed as often as needed. They should last through at least a week's worth of experiments.
Caution should be used when changing solutions so as not to introduce air bubbles. It is a good idea to
open both sides of the system to atmosphere giving zero flow. It may be possible to change solution
under low suction (if necessary to maintain a seal) without introducing air bubbles if one works quickly.
Again, individual factors related to the whole experiment will determine this. It is a good idea to practice
changes under suction to see if the outcome is satisfactory.
Page 13 of 20
Patch perfusion Setup with unprepared Quartz Capillary
Patch perfusion Setup with prepared Quartz Capillary
Page 14 of 20
Helpful Hints
• Try to use filtered solutions for perfusion. De-gas all solutions slightly before use by gentle warming.
• Either pressure or vacuum can be monitored at a time. It is not possible to monitor both
simultaneously. The mode switch will not introduce bias to the pressure measurement.
• The 2PK+ is designed to measure down to +/- 0.1 mmHg. It can be affected by changes in
atmospheric pressure, i.e. weather or wind conditions. As a rule we claim that the pressure
measurement is accurate to 0.5mm/Hg. As the measuring capability is 0.1mm/Hg in fact greater
accuracy is possible, but we feel that measurement of less than 0.5mm/Hg should be used as a
reference.
• The advent of the positive pressure vessel may obviate the need to pull the quartz capillary to a fine tip
-- this should be determined empirically. Bear in mind that the smaller the tip of the perfusion capillary,
the closer to the tip of the recording pipette it can be placed.
• Shutting off the instrument does not stop gas flow. Turn off gas at the source when the unit is not in
use.
• Perfusion in both whole cell and pipette mode can take seconds to minutes. Times can be changed by
altering perfusion capillary tip size, pressure/vacuum settings and PE tubing length. Other factors like
cell size, type, and temperature have a great effect on perfusion speeds.
• Dozens of perfusions are possible before backfilling the pipette to the suction line. Longer/larger
pipettes should be used for long and complex experiments. ALA Scientific can provide perfusion
O
pipette holders with dual 30 ports but we do not recommend them since an additional tube in the
recording pipette will add more noise.
• Be sure to keep the pipette holder clean. Once a week it should be washed in distilled water and
methanol and dried in a desiccator. The same is true for the positive pressure vessel. Both items are
made of polycarbonate, so that frequent cleaning is necessary for low noise recordings.
• To perfuse sharp electrodes we recommend using the whole cell approach. The sharp electrode may
be less efficient for actual whole cell perfusion due to its small tip diameter. However, it should be
possible to perfuse the electrode.
Page 15 of 20
Limited Warranty
ALA Scientific Instruments agrees to warranty this product for one year from the date of shipment. Said
warranty covers all parts and labor necessary to remedy defects in workmanship and/or materials.
Coverage is limited to repair or replacement of parts. Items not covered under this limited warranty
include external tubing, pipette holders, pressure vessel, butane torch, connectors, internal filters, luer
valves and all disposable parts. All units returned to ALA Scientific Instruments for repair, whether under
warranty or not, must be returned freight prepaid. Freight collect will be refused and will result in longer
repair times.
ALA Scientific Instruments Inc. assumes no liability for damage to or resulting from the use of this product
including damage to other equipment, personal property and persons using this instrument. The user is
responsible for using this instrument in accordance with this manual for the intended purpose of internal
pipette and whole cell perfusion. Usage that is inconsistent with this intent may result in forfeiture of
warranty. Please consult ALA Scientific Instruments if considering alternative applications.
Please note: It is the buyer's responsibility to inspect this instrument upon receipt for possible damage
that has resulted from shipping and to report any claim to the carrier within three business days.
If the unit is damaged in shipment it is the buyer's responsibility to file a claim with the carrier. ALA
Scientific Instruments will assist as much as possible.
This instrument is designed for non-clinical use only!
Page 16 of 20
Diagrams
Block Diagram
Valve Modes to be Avoided
Page 17 of 20
Specifications
Wall Power Supply Output
9VDC/100mA (center = +)
Power Consumption
0.08W
MAX. Input Pressure
50 PSI (345 kPa)
MIN. Input Pressure
15 PSI (100kPa)
MIN. Output Pressure
0
MAX. Output Pressure
50 mmHg (6.66kPa)
MAX. Output Vacuum
-40 mmHg (-5.33kPa)
Input Pressure Connector
1/8in ID tube barb
Output Pressure Connectors
Luer lock
Dimensions
16 cm x 15.8 cm x 8.5 cm (LWH)
Weight
1 kg
Fuse
100mA (Located inside generator)
This instrument complies with all applicable standards for light industrial machinery at the European
Union designated by the CE mark
NOTE on Pressure Supply:
A pressure pump producing 2-4ATM of pressure may be substituted for the air/nitrogen cylinder. The
output of the pump must be filtered to be free of condensation so that moisture will not get into the 2PK+
and harm the regulators and pressure transducer.
Page 18 of 20
Pressure Conversion Table
Conversion table for millimeters of mercury to kilopascals
kPa
mmHg
kPa
mmHg
mHg
1
1.5
2
2.5
3
3.5
4
4.5
5
5.5
6
6.5
7
7.5
8
8.5
9
9.5
10
10.5
11
11.5
12
12.5
13
13.5
14
14.5
15
15.5
16
16.5
17
17.5
18
18.5
19
19.5
20
20.5
21
21.5
22
22.5
23
23.5
24
24.5
25
25.5
26
26.5
27
27.5
.1333
.1999
.2666
.3333
.3999
.4666
.5333
.5999
.6666
.7333
.7999
.8666
.9333
.9999
1.067
1.133
1.2
1.267
1.333
1.4
1.467
1.533
1.6
1.667
1.733
1.8
1.867
1.933
2
2.066
2.133
2.2
2.267
2.333
2.4
2.466
2.533
2.6
2.666
2.733
2.8
2.866
2.933
3
3.066
3.133
3.2
3.266
3.333
3.4
3.466
3.533
3.6
2.796
28
28.5
29
29.5
30
30.5
31
31.5
32
32.5
33
33.5
34
34.5
35
35.5
36
36.5
37
37.5
38
38.5
39
39.5
40
40.5
41
41.5
42
42.5
43
43.5
44
44.5
45
45.5
46
46.5
47
47.5
48
48.5
49
49.5
50
50.5
51
51.5
52
52.5
53
53.5
54
54.5
3.733
3.8
3.866
3.933
4
4.066
4.133
4.2
4.266
4.333
4.4
4.466
4.533
4.6
4.666
4.733
4.8
4.866
4.933
5
5.066
5.133
5.2
5.266
5.333
5.4
5.466
5.533
5.6
5.666
5.733
5.8
5.866
5.933
6
6.066
6.133
6.2
6.266
6.333
6.4
6.466
6.533
6.6
6.666
6.733
6.8
6.866
6.933
7
7.066
7.133
7.2
7.266
Page 19 of 20
55
55.5
56
56.5
57
57.5
58
58.5
59
59.5
60
60.5
61
61.5
62
62.5
63
63.5
64
64.5
65
65.5
66
66.5
67
67.5
68
68.5
69
69.5
70
70.5
71
71.5
72
72.5
73
73.5
74
74.5
75
75.5
76
76.5
77
77.5
78
78.5
79
79.5
80
80.5
81
81.5
kPa
7.333
7.4
7.466
7.533
7.6
7.666
7.733
7.8
7.866
7.933
8
8.066
8.133
8.199
8.266
8.333
8.399
8.466
8.533
8.6
8.666
8.733
8.8
8.866
8.933
9
9.066
9.133
9.2
9.266
9.333
9.4
9.466
9.533
9.6
9.666
9.733
9.8
9.866
9.932
10
10.066
10.132
10.2
10.266
10.332
10.4
10.466
10.532
10.6
10.666
10.732
10.8
10.866
Reference Articles
•
Tang JM, Wang J, Quandt FN, Eisenberg RS, "Perfusing Pipettes,"
Pflugers Archives (1990) 41 :347-350
Additional Articles:
2+
•
Song Y, Simard JM, " -Adrenoceptor stimulation activates large conductance Ca -activated K+
channels in smooth muscle cells from basilar artery of guinea pig," Pflugers Arch, Eur J Physiol
(1995) 430:984-993
•
Practical electrophysiological methods: a guide for invitro studies in vertebrate neurobiology.
Editors: Kettenmann H. & Grantyn R. Wiley Liss, inc. 1992 (Chapter 4.11 intracellular Perfusion
by Patch Electrodes, Hescheller J, Kameyama M, Speicher R. 241 245
(highly
recommended)
Page 20 of 20
Was this manual useful for you? yes no
Thank you for your participation!

* Your assessment is very important for improving the work of artificial intelligence, which forms the content of this project

Download PDF

advertisement