SUTTER INSTRUMENT P-97 Operation Manual

OPERATION MANUAL

P

P----97

PP

Flaming/Brown

Flaming/BrownFlaming/Brown

Micropipette

MicropipetteMicropipette

Puller

PullerPuller

Rev. 2.03 – DOM (20091301)

97

9797

P

P----97

PP

Flaming/Brown Micropipette Puller

Flaming/Brown Micropipette PullerFlaming/Brown Micropipette Puller

97

9797

Operation Manual

(Rev. 2.03 – DOM (20091301))

Sutter Instrument Company

One Digital Drive Novato, CA 94949

Voice: 415-883-0128 Web: www.sutter.com Fax: 415-883-0572 Email: info@sutter.com



P-97 FLAMING/BROWN MICROPIPETTE PULLER OPERATION MANUAL – REV. 2.03 – DOM (20091301)

TABLE OF CONTENTS

TABLE OF CONTENTSTABLE OF CONTENTS

1. GENERAL INFORMATI

1. GENERAL INFORMATION

1. GENERAL INFORMATI1. GENERAL INFORMATI

1.1 T

ECHNICAL SUPPORT

1.2 W

HAT’S NEW ON THE

2. SAFETY WARNINGS A

2. SAFETY WARNINGS AND PRECAUTI

2. SAFETY WARNINGS A2. SAFETY WARNINGS A

2.1 M

AIN FUSE

2.2 A

VOIDING ELECTRICAL SHOCK OR FIRE INJURY

2.3 B

ACK INJURY PREVENTION

3. PRECAUTIONS

3. PRECAUTIONS ................................

3. PRECAUTIONS3. PRECAUTIONS

3.1 P

RECAUTIONS DURING OPERATION

3.2 ON H

4. GLASS CAPILLARY &

4. GLASS CAPILLARY & HEATING FILAMENT SP

4. GLASS CAPILLARY &4. GLASS CAPILLARY &

4.1 G

LASS CAPILLARY

4.2 H

EATING FILAMENT

.........................................................................................................................................3

................................................................

ANDLING MICROPIPETTES

ON ................................

................................................................

ONON

................................................................

........................................................................................................................1

P-97?..............................................................................................................1

ND PRECAUTIONS

ND PRECAUTIND PRECAUTI

ONS................................

................................................................

ONSONS

................................................................

.............................................................................3

..............................................................................................................3

................................................................

.................................................................................................5

.......................................................................................................5

HEATING FILAMENT SPECIFICATIONS

HEATING FILAMENT SP HEATING FILAMENT SP

..............................................................................................................................7

..........................................................................................................................7

...........................................

................................................................

................................................

................................................................

...............................................................

................................................................

ECIFICATIONS................................

ECIFICATIONSECIFICATIONS

..........................................

................................................................

...............................5555

..............................................................

................3333

................................

...........1111

......................

..........7777

....................

5. UNPACKING

5. UNPACKING................................

5. UNPACKING5. UNPACKING

6. SETTING UP

6. SETTING UP................................

6. SETTING UP6. SETTING UP

6.1 L

INE POWER (MAINS

7. FRONT PANEL

7. FRONT PANEL................................

7. FRONT PANEL7. FRONT PANEL

7.1 C

ONTROLS

7.2 D

ISPLAY

8. MECHANICAL DESCRI

8. MECHANICAL DESCRIPTION (PULLER ANATOM

8. MECHANICAL DESCRI8. MECHANICAL DESCRI

8.1 S

OME BASIC INFORMATION

8.2 A

IR COOLING SYSTEM

8.3 H

EATING ASSEMBLY

8.4 U

PPER PULLEY ASSEMBLY

8.5 C

ABINET

8.6 E

LECTRONICS

9. FIRST TIM

9. FIRST TIME USE

9. FIRST TIM9. FIRST TIM

10. PROGRAMS

10. PROGRAMS................................

10. PROGRAMS10. PROGRAMS

10.1 P

10.2 P

10.3 P

10.4 D

10.5 S

10.6 V

10.7 C

10.8 E

ROGRAM STRUCTURE ROGRAM LINE PULL CYCLE PARAMETERS ULL CYCLE

EFAULT CONFIGURATION

ELECTING A PROGRAM

IEWING A PROGRAM LEARING A PROGRAM FROM MEMORY DITING A PROGRAM

................................................................

....................................

................................................................

..................................

................................................................

)......................................................................................................................11

................................................................

..............................................................

................................................................

..............................13

............................................................

........................................................................................................................................13

............................................................................................................................................14

PTION (PULLER ANATOMY)

PTION (PULLER ANATOMPTION (PULLER ANATOM

Y)................................

............................................................

Y)Y)

................................................................

............................17

........................................................

.............................................................................................................17

.....................................................................................................................17

........................................................................................................................18

..............................................................................................................20

...........................................................................................................................................21

...................................................................................................................................21

E USE ................................

E USEE USE

................................................................

..........................................................

................................................................

..........................23

....................................................

..................................

................................................................

..................................................................................................................25

................................................................................25

...................................................................................................................................26

...........................................................................................................27

................................................................................................................27

....................................................................................................................28

.......................................................................................28

.....................................................................................................................28

10.8.1 Entering a New Program......................................................................................................28

10.8.2 Editing an Existing Program................................................................................................29

....9999

........

..11

11

....

1111

13

1313

17

1717

23

2323

..25

25

....

2525

P-97 FLAMING/BROWN MICROPIPETTE PULLER OPERATION MANUAL – REV. 2.03 – DOM (20091301)

11. SOFTWARE CONTROL

11. SOFTWARE CONTROL FUNCTIONS

11. SOFTWARE CONTROL11. SOFTWARE CONTROL

11.1 RAMP TEST < 1 >......................................................................................................................31

11.2 CHANGE AIR PRESSURE < 2 >.............................................................................................32

11.3 AIR MODE < 3 >.........................................................................................................................33

11.4 AIR TIME AT START OF PULL < 4 > ...................................................................................33

11.5 AIR TIME AT END OF PULL < 5 >........................................................................................34

11.6 RESET TIME AND DATE < 6 > ..............................................................................................34

11.7 WRITE-PROTECT THE PROGRAM < 7 >............................................................................34

11.8 MEMORY TEST < 8 >...............................................................................................................34

12. PULLING PIPETTES

12. PULLING PIPETTES................................

12. PULLING PIPETTES12. PULLING PIPETTES

12.1 P

12.2 N

13. PARAMETER ADJUST

13. PARAMETER ADJUSTMENT

13. PARAMETER ADJUST13. PARAMETER ADJUST

13.1 G

13.2 M

13.3 P

13.4 S

13.5 T

13.6 F

ROCEDURES

OTES ON PROGRAM OPERATION

ENERAL INFORMATION

ICROPIPETTE/MICROINJECTION NEEDLE FABRICATION

13.2.1 HEAT ......................................................................................................................................37

13.2.2 PULL Strength.......................................................................................................................37

13.2.3 VELOCITY (Trip Point)........................................................................................................38

13.2.4 TIME Mode (Cooling)............................................................................................................38

13.2.5 Delay Mode (Cooling).............................................................................................................38

13.2.6 PRESSURE Adjustment.......................................................................................................39

13.2.7 Filament Width ......................................................................................................................39

ATCH PIPETTE FABRICATION

13.3.1 HEAT ......................................................................................................................................40

13.3.2 PULL Strength.......................................................................................................................40

13.3.3 VELOCITY (Trip Point)........................................................................................................40

13.3.4 TIME Mode (Cooling)............................................................................................................41

13.3.5 PRESSURE ............................................................................................................................41

TEP-BY-STEP PATCH PIPETTE PROGRAMMING

ECHNICAL TIPS FOR PULLING MICROPIPETTES

13.5.1 Regulating the Time it takes to pull a Sharp Pipette ........................................................43

13.5.1.1 Pipette Position..........................................................................................................................................................43

13.5.2 Regulating the Length and Tip Size of a Sharp Pipette....................................................43

13.5.2.1 Filament Width..........................................................................................................................................................43

13.5.2.2 Air Flow.......................................................................................................................................................................43

13.5.2.3 DELAY Mode of Active Cooling..............................................................................................................................44

IRE POLISHING

..................................................................................................................................35

FUNCTIONS................................

FUNCTIONS FUNCTIONS

................................................................

MENT ................................

MENTMENT

............................................................................................................................44

................................................................

...............................................................................................................37

.....................................................................................................40

................................................................

................................................................................................36

................................................................

.........................................................................41

.......................................................................43

......................................................

................................................................

..................................................

................................................................

...................................

................................................................

.........................................................37

......................31

............................................

..................35

....................................

31

3131

35

3535

...37

37

......

3737

14. HEATING FILAMENT

14. HEATING FILAMENTSSSS ................................

14. HEATING FILAMENT14. HEATING FILAMENT

14.1 G

14.2 T

14.3 B

15. MAINTENANCE

15. MAINTENANCE................................

15. MAINTENANCE15. MAINTENANCE

ENERAL INFORMATION

ROUGH FILAMENT

14.2.1 Positioning..............................................................................................................................47

14.2.2 Geometry.................................................................................................................................47

OX FILAMENT

14.3.1 Positioning..............................................................................................................................49

14.3.2 Geometry.................................................................................................................................49

P-97 FLAMING/BROWN MICROPIPETTE PULLER OPERATION MANUAL – REV. 2.03 – DOM (20091301)

..............................................................................................................................48

................................................................

...............................................................................................................47

.......................................................................................................................47

................................................................

..........................................................

................................................................

.............................................

................................................................

..........................51

....................................................

.............47

..........................

47

4747

51

5151

15.1 C

15.2 H

15.3 P

15.4 R

16. TROUBLESHOOTING

16. TROUBLESHOOTING................................

16. TROUBLESHOOTING16. TROUBLESHOOTING

16.1 C

16.2 C

16.3 T

LEANING

EATING FILAMENT REPLACEMENT

15.2.1 Systematic Filament Replacement.......................................................................................51

15.2.2 Air jet Position........................................................................................................................51

15.2.3 Positioning the Filament in Relation to the Glass Capillary ............................................51

ULLEY ADJUSTMENT EGENERATION OF DRIERITE GRANULES

ONTROLLING PIPETTE TIP SHAPES

16.1.1 Problem: What type of glass should be used?.....................................................................57

16.1.2 Problem: The resistance of pulled pipettes is too low. How can a higher-resistance

pipette be pulled?...............................................................................................................................57

16.1.3 Problem: Okay, but how can tips be made even smaller?.................................................57

16.1.4 Problem: How can the size of a patch-pipette be increased?.............................................58

16.1.5 Problem: Patch-pipette tips vary in size from pull to pull.................................................58

16.1.6 Problem: An injection pipette with a 1mm tip that is 20 to 50mm long needs to be

formed. How can this be done?.......................................................................................................58

16.1.7 Problem: The electrodes are bent. How can they be made to pull straight?..................58

16.1.8 Problem: One electrode is much longer than the other electrode....................................58

16.1.9 Problem: The shape and resistance of the pipette changes from pull to pull..................59

ONTROLLER PROBLEMS

16.2.1 Problem: The filament does not light up when pull is pressed........................................59

16.2.2 Problem: Display blank, fan not on, or locked up/overlap................................................60

16.2.3 Problem: Display shows a row of blocks.............................................................................60

16.2.4 Problem: Displayed program values are not correct.........................................................60

ECHNICAL SUPPORT

.......................................................................................................................................51

...........................................................................................51

...................................................................................................................53

...................................................................................54

................................................................

..............................................................................................................59

....................................................................................................................60

................................................................

...........................................................................................57

................................................

................................................................

................57

................................

57

5757

APPENDIX A. FUSE REP

APPENDIX A. FUSE REPLACEMENT

APPENDIX A. FUSE REPAPPENDIX A. FUSE REP

APPENDIX B. LIMITED

APPENDIX B. LIMITED WARR

APPENDIX B. LIMITED APPENDIX B. LIMITED

APPENDIX C. DISCLAIM

APPENDIX C. DISCLAIMER

APPENDIX C. DISCLAIMAPPENDIX C. DISCLAIM

APPENDIX D. TECHNICA

APPENDIX D. TECHNICAL SPECIFICATIONS

APPENDIX D. TECHNICAAPPENDIX D. TECHNICA

APPENDIX E. DRIERITE

APPENDIX E. DRIERITE MATERIAL SAFETY DAT

APPENDIX E. DRIERITEAPPENDIX E. DRIERITE

INDEX

INDEX................................

INDEXINDEX

TABLE OF FIGURES

TABLE OF FIGURESTABLE OF FIGURES

Figure 6-1. P-97 Cabinet (rear view)......................................................................................................11

Figure 6-2. P-97 Cabinet (end view, left). ..............................................................................................11

Figure 6-3. Power connection..................................................................................................................11

Figure 7-1. P-97 Front Panel...................................................................................................................13

Figure 7-2. Program Display...................................................................................................................14

Figure 8-1. P-97 Base Plate (Top view, cover removed).......................................................................17

................................................................

LACEMENT................................

LACEMENTLACEMENT

WARRANTY

WARRWARR

ANTY................................

ANTYANTY

ER................................

................................................................

ERER

................................................................

L SPECIFICATIONS................................

L SPECIFICATIONSL SPECIFICATIONS

MATERIAL SAFETY DATA SHEET

MATERIAL SAFETY DAT MATERIAL SAFETY DAT

................................................................

...........................................................

................................................................

............................................................

................................................................

A SHEET................................

A SHEETA SHEET

...........................61

......................................................

............................63

........................................................

............................................

................................................................

...........................................

................................................................

..................................................

................................................................

.................................................

................................................................

............65

........................

...........67

......................

..................69

....................................

.................71

..................................

61

6161

63

6363

65

6565

67

6767

69

6969

71

7171

P-97 FLAMING/BROWN MICROPIPETTE PULLER OPERATION MANUAL – REV. 2.03 – DOM (20091301)

Figure 8-2. Filament Block Assembly. ...................................................................................................18

Figure 8-3. Upper cable pulley assembly................................................................................................20

Figure 9-1. Left puller bar. ......................................................................................................................23

Figure 10-1. Power-On Display...............................................................................................................27

Figure 10-2. P-97 Program Display........................................................................................................27

Figure 10-3. Clear Program Display.......................................................................................................28

Figure 11-1. Access to Control Functions..............................................................................................31

Figure 12-1. Pull Cycle Report................................................................................................................35

Figure 12-2. Sample program..................................................................................................................36

Figure 12-3. Pull cycle report (multiple-loops)......................................................................................36

Figure 13-1. Side view of fire polishing spacer. .....................................................................................45

Figure 13-2. Fire Polish Spacer in Puller Bar. ......................................................................................45

Figure 14-1. End view of trough filament and glass.............................................................................47

Figure 14-2. End view of box filament. ..................................................................................................48

Figure 15-1. V-Groove bearings and puller bars...................................................................................51

Figure 15-2. Filament alignment............................................................................................................52

Figure 15-3. Micropipette shapes............................................................................................................53

Figure 15-4. Pulley Adjustment..............................................................................................................54

Figure 16-1. Power entry module. ..........................................................................................................61

Figure 16-2. Fuse holder..........................................................................................................................61

TABLE OF TABLES

TABLE OF TABLESTABLE OF TABLES

Table 10-1. Standard Factory Configuration.........................................................................................27

Table 11-1. Control Functions.................................................................................................................31

Table 11-2. Maximum heat and recommended starting values for different filament shapes........32

Table 14-1. Trough filament sizes. .........................................................................................................48

Table 14-2. Box filament sizes.................................................................................................................49

P-97 FLAMING/BROWN MICROPIPETTE PULLER OPERATION MANUAL – REV. 2.03 – DOM (20091301)

1

1.

1. G E NERAL I NF ORMATI ON

GEN E RA L INFO RM ATION

1.1 .

GEN E RA L INFO RM ATIONGEN E RA L INFO RM ATION

The P-97 can fabricate pipettes for use in intracellular recording, patch clamping, transferring (ICSI, ES Cells), microinjection, aspiration, and microperfusion. Realizing the full potential of this instrument is dependent on a complete understanding of the way it implements the pulling process. To this end, we urge that this manual be read in its entirety. To aid in understanding the function of the instrument, sample programs are already loaded in memory (as discussed in subsequent material).

The Model P-97 Flaming/Brown Micropipette Puller combines a proven pulling technology with programmability to produce a very versatile instrument. The pulling mechanism is derived from the P-77/P-80 series of pullers, which have demonstrated the ability to pull a complete range of pipette profiles. Added to this mechanism is the ability to program different pulling sequences; thus, allowing ease of use for pulling a multiplicity of pipettes on one device.

The P-97 is a ‘velocity sensing’ puller. This patented feature allows the puller to indirectly sense the viscosity of the glass, giving the P-97 the ability to pull pipettes from all glasses except quartz. Even difficult to pull formulations, such as aluminosilicate glasses, are handled with relative ease.

Throughout this manual reference will be made to the size of the glass tubing used to pull micropipettes. The convention used here for describing the outside diameter (O.D.) and inside diameter (I.D.) is as follows: O.D. x I.D. These dimensions will always be given in millimeters (mm). See our catalog or visit

www.sutter.com

to find a wide selection of glass

capillaries available for purchase.

1.1

1.1 Technical Support

Technical Support

1.11.1

Technical SupportTechnical Support

Unlimited technical support is provided by Sutter Instrument Company at no charge to our customers. Our technical support staff is available between the hours of 8:00 AM and 5:00 PM (Pacific Standard Time) at (415) 883-0128. You may also Email your queries to

info@sutter.com.

1.2

1.2 What’s New on the P

What’s New on the P----97?

1.21.2

What’s New on the PWhat’s New on the P

97?

97?97?

For users familiar with the P-80PC and/or P-87 pullers, the P-97 provides the advanced features incorporated into those instruments plus:



Humidity controlled chamber surrounding the heating filament. Glass is now pulled in a

Humidity controlled chamber surrounding the heating filament. Glass is now pulled in a Humidity controlled chamber surrounding the heating filament. Glass is now pulled in a constant humidity environment to minimize environmental effects on

constant humidity environment to minimize environmental effects on pipette

constant humidity environment to minimize environmental effects onconstant humidity environment to minimize environmental effects on reproducibility.

reproducibility.

reproducibility.reproducibility.



Vacuum fluorescent display for improved readability

Vacuum fluorescent display for improved readabilityVacuum fluorescent display for improved readability

pipette

pipette pipette



25% increase in available power

25% increase in available power25% increase in available power



Programmable air pressure

Programmable air pressureProgrammable air pressure



Time of pull displayed at the end of each pull sequence

Time of pull displayed at the end of each pull sequenceTime of pull displayed at the end of each pull sequence



Write protection to prevent programs from being corrupted

Write protection to prevent programs from being corruptedWrite protection to prevent programs from being corrupted

P-97 FLAMING/BROWN MICROPIPETTE PULLER OPERATION MANUAL – REV. 2.03 – DOM (20091301)

2



Displa

Displayed date/time of last program edit

DisplaDispla



Increased memory capable of storing 100 programs

Increased memory capable of storing 100 programsIncreased memory capable of storing 100 programs



Switchable cooling mode to facilitate easy transfer of programs from both P

Switchable cooling mode to facilitate easy transfer of programs from both P----80PC and P

Switchable cooling mode to facilitate easy transfer of programs from both PSwitchable cooling mode to facilitate easy transfer of programs from both P 87 pullers

87 pullers

87 pullers87 pullers

yed date/time of last program edit

yed date/time of last program edityed date/time of last program edit

80PC and P----

80PC and P80PC and P

P-97 FLAMING/BROWN MICROPIPETTE PULLER OPERATION MANUAL – REV. 2.03 – DOM (20091301)

3

2.

2. SA FE TY WARN I NG S AND

SAF E TY WARNI NG S AND PRECA UT IO NS

2.2 .

SAF E TY WARNI NG S AND SAF E TY WARNI NG S AND

2.1

2.1 Main Fuse

Main Fuse

2.12.1

Main FuseMain Fuse





Repl

Replace fuse only with the same type and rating:

ace fuse only with the same type and rating:

Repl Repl

ace fuse only with the same type and rating:ace fuse only with the same type and rating:

Type:

Type: 5 x 20 mm glass tube, Medium Time Delay (IEC

Type:Type:

Rating:

Rating: 3A 250V (3 Amps, 250 Volts)

Rating:Rating:

Examples:

Examples: Bussmann GMC

Examples:Examples:

A spare fuse is provided, located in the power input module. Please refer to

A spare fuse is provided, located in the power input module. Please refer to Appendix A

A spare fuse is provided, located in the power input module. Please refer to A spare fuse is provided, located in the power input module. Please refer to Fuse

Fuse Replacement

Replacement for more details on fuse ratings and for instructions on how to change

FuseFuse

ReplacementReplacement

the fuse.

the fuse. the fuse.

2.2

2.2 Avoiding Electrical Shock or Fire Injury

Avoiding Electrical Shock or Fire Injury

2.22.2

Avoiding Electrical Shock or Fire InjuryAvoiding Electrical Shock or Fire Injury

5 x 20 mm glass tube, Medium Time Delay (IEC 60127

5 x 20 mm glass tube, Medium Time Delay (IEC5 x 20 mm glass tube, Medium Time Delay (IEC

Time Lag, RoHS compliant.

Time Lag, RoHS compliant.Time Lag, RoHS compliant.

3A 250V (3 Amps, 250 Volts)

3A 250V (3 Amps, 250 Volts) 3A 250V (3 Amps, 250 Volts)

Bussmann GMC----3A or GMC

Bussmann GMCBussmann GMC

for more details on fuse ratings and for instructions on how to change

for more details on fuse ratings and for instructions on how to change for more details on fuse ratings and for instructions on how to change

PRECA UTION S

PRECA UTION SPRE CA UTION S

3A or GMC----3333----R; or Littelfus

3A or GMC3A or GMC

R; or Littelfuse 239 003 or 239 003.P)

R; or LittelfusR; or Littelfus

60127----2, Sheet III) or

6012760127

e 239 003 or 239 003.P)

e 239 003 or 239 003.P)e 239 003 or 239 003.P)

2, Sheet III) or

2, Sheet III) or 2, Sheet III) or

Appendix A

Appendix AAppendix A



To prevent fire or shock hazard do not expose the unit to rain or moisture.

To prevent fire or shock hazard do not expose the unit to rain or moisture.To prevent fire or shock hazard do not expose the unit to rain or moisture.



Use only a properly grounded power source; appropriate

Use only a properly grounded power source; appropriately rated for use with this

Use only a properly grounded power source; appropriateUse only a properly grounded power source; appropriate instrument. Always use the grounded power supply cord set provided to connect the unit

instrument. Always use the grounded power supply cord set provided to connect the unit

instrument. Always use the grounded power supply cord set provided to connect the unit instrument. Always use the grounded power supply cord set provided to connect the unit to the grounded outlet (3

to the grounded outlet (3----prong). This is required to protect you from injury in the event

to the grounded outlet (3to the grounded outlet (3 that an electrical hazard occurs.

that an electrical hazard occurs.

that an electrical hazard occurs.that an electrical hazard occurs.



Do not disassemb

Do not disassemble the unit. Refer servicing to Sutter Instrument Company.

Do not disassembDo not disassemb

2.3

2.3 Back Injury Prevention

Back Injury Prevention

2.32.3

Back Injury PreventionBack Injury Prevention

To avoid injuring your back or limbs it is recommended that you do not attempt to lift this

To avoid injuring your back or limbs it is recommended that you do not attempt to lift this To avoid injuring your back or limbs it is recommended that you do not attempt to lift this instrument by yourself. The P

instrument by yourself. The P----97 Micropipette Puller weighs in excess of 18 kg (

instrument by yourself. The Pinstrument by yourself. The P and should be moved by TWO (2) people.

and should be moved by TWO (2) people.

and should be moved by TWO (2) people.and should be moved by TWO (2) people.

le the unit. Refer servicing to Sutter Instrument Company.

le the unit. Refer servicing to Sutter Instrument Company.le the unit. Refer servicing to Sutter Instrument Company.

prong). This is required to protect you from injury in the event

prong). This is required to protect you from injury in the event prong). This is required to protect you from injury in the event

97 Micropipette Puller weighs in excess of 18 kg (over 39 lb)

97 Micropipette Puller weighs in excess of 18 kg (97 Micropipette Puller weighs in excess of 18 kg (

ly rated for use with this

ly rated for use with this ly rated for use with this

over 39 lb)

over 39 lb) over 39 lb)

P-97 FLAMING/BROWN MICROPIPETTE PULLER OPERATION MANUAL – REV. 2.03 – DOM (20091301)

5

3.

3. PR EC AU TI ONS

PRE C AU TI ONS

3.3 .

PRE C AU TI ONSPRE C AU TI ONS

3.1

3.1 Precautions during Operation

Precautions during Operation

3.13.1

Precautions during OperationPrecautions during Operation

Failure to comply with any of the following precautions may damage this device.

Failure to comply with any of the following precautions may damage this device. Failure to comply with any of the following precautions may damage this device.



Operate the P

Operate the P----97 using the indicated line voltage.

Operate the POperate the P



The P

The P----97 is designed for operation in a laboratory environment (pollution degree II) and

The PThe P at temperatures between 5°C

at temperatures between 5°C ---- 40°C.

at temperatures between 5°C at temperatures between 5°C



The P

The P----97 is designed for connection to a standard laboratory power outlet (overvoltage

The PThe P Category II) with main supply voltage fluct

Category II) with main supply voltage fluctuations not to exceed ± 10% of the normal

Category II) with main supply voltage fluctCategory II) with main supply voltage fluct voltage.

voltage.

voltage.voltage.



This unit is not designed for operation, nor has it been tested for safety, at altitudes

This unit is not designed for operation, nor has it been tested for safety, at altitudes This unit is not designed for operation, nor has it been tested for safety, at altitudes above 2000 meters (6562 feet).

above 2000 meters (6562 feet).

above 2000 meters (6562 feet).above 2000 meters (6562 feet).



This unit was designed to operate at maximum relative humidity of 80% for temperature

This unit was designed to operate at maximum relative humidity of 80% for temperatures

This unit was designed to operate at maximum relative humidity of 80% for temperatureThis unit was designed to operate at maximum relative humidity of 80% for temperature up to 31°C decreasing linearly to 50% relative humidity at 40°C.

up to 31°C decreasing linearly to 50% relative humidity at 40°C.

up to 31°C decreasing linearly to 50% relative humidity at 40°C.up to 31°C decreasing linearly to 50% relative humidity at 40°C.

97 is designed for operation in a laboratory environment (pollution degree II) and

97 is designed for operation in a laboratory environment (pollution degree II) and 97 is designed for operation in a laboratory environment (pollution degree II) and

97 is designed for connection to a standard laboratory power outlet (overvoltage

97 is designed for connection to a standard laboratory power outlet (overvoltage 97 is designed for connection to a standard laboratory power outlet (overvoltage

97 using the indicated line voltage.

97 using the indicated line voltage. 97 using the indicated line voltage.

40°C.

40°C. 40°C.

uations not to exceed ± 10% of the normal

uations not to exceed ± 10% of the normal uations not to exceed ± 10% of the normal

s

s s



Operate only in a location where there is a free flow of fresh air on all sides. The fan

Operate only in a location where there is a free flow of fresh air on all sides. The fan Operate only in a location where there is a free flow of fresh air on all sides. The fan draws air in through the vents on the sides and exhausts out both ends of the heat sink.

draws air in through the vents on the sides and exhausts out both ends of the heat sink.

draws air in through the vents on the sides and exhausts out both ends of the heat sink. draws air in through the vents on the sides and exhausts out both ends of the heat sink. NEVER ALL

NEVER ALLOW THE FREE FLOW OF AIR TO BE RESTRICTED.

NEVER ALLNEVER ALL



To avoid burns do not touch the heating filament, the brass clamps holding the filament,

To avoid burns do not touch the heating filament, the brass clamps holding the filament, To avoid burns do not touch the heating filament, the brass clamps holding the filament, or the heated ends of glass pipettes that have been pulled.

or the heated ends of glass pipettes that have been pulled.

or the heated ends of glass pipettes that have been pulled.or the heated ends of glass pipettes that have been pulled.



Only use Sutter Instrument Company replacement heating filaments.

Only use Sutter Instrument Company replacement heating filaments.Only use Sutter Instrument Company replacement heating filaments.

3.2

3.2 On Handling Micropipettes

On Handling Micropipettes

3.23.2

On Handling MicropipettesOn Handling Micropipettes

Failure to comply with any of the following precautions may result in injury to the users of

Failure to comply with any of the following precautions may result in injury to the users of Failure to comply with any of the following precautions may result in injury to the users of this device as well as those working in the general area near the device.

this device as well as those working in the general area near the device.

this device as well as those working in the general area near the device.this device as well as those working in the general area near the device.



The micropipettes created using t

The micropipettes created using this instrument are very sharp and relatively fragile.

The micropipettes created using tThe micropipettes created using t Contact with the pulled micropipette tips, therefore, should be avoided to prevent

Contact with the pulled micropipette tips, therefore, should be avoided to prevent

Contact with the pulled micropipette tips, therefore, should be avoided to prevent Contact with the pulled micropipette tips, therefore, should be avoided to prevent accidentally impaling yourself.

accidentally impaling yourself.

accidentally impaling yourself. accidentally impaling yourself.



Always dispose of micropipettes by placing them into a well

Always dispose of micropipettes by placing them into a well----marked, spill

Always dispose of micropipettes by placing them into a wellAlways dispose of micropipettes by placing them into a well container.

container.

container.container.



Use only with capillary glass (tubing) recommended by Sutter Instrument Company in

Use only with capillary glass (tubing) recommended by Sutter Instrument Company in Use only with capillary glass (tubing) recommended by Sutter Instrument Company in the following section of this manual (

the following section of this manual (4.1

the following section of this manual (the following section of this manual (

OW THE FREE FLOW OF AIR TO BE RESTRICTED.

OW THE FREE FLOW OF AIR TO BE RESTRICTED.OW THE FREE FLOW OF AIR TO BE RESTRICTED.

his instrument are very sharp and relatively fragile.

his instrument are very sharp and relatively fragile. his instrument are very sharp and relatively fragile.

marked, spill----proof “sha

marked, spillmarked, spill

4.1).

).

4.14.1

).).

proof “sharps”

proof “shaproof “sha

rps”

rps” rps”

P-97 FLAMING/BROWN MICROPIPETTE PULLER OPERATION MANUAL – REV. 2.03 – DOM (20091301)

7

4.

4. G L AS S CAPI LL A RY & HE

GLA S S CAPI L LA RY & H EATIN G FI LA MENT S PE CI

4.4 .

GLA S S CAPI L LA RY & H EGLA S S CAPI LL A RY & HE

4.1

4.1 Glass Capillary

Glass Capillary

4.14.1

Glass CapillaryGlass Capillary

The P-97 micropipette puller is designed for use with aluminosilicate, borosilicate or other lower melting-point glass tubing or rod ranging from 0.6 to 3.0 mm in diameter. This instrument does not pull quartz glass. Examples of the specific types and sizes of glass that can be used with the P-97 are listed in the Sutter Instrument Company catalogue that was included with this instrument or can be viewed on Sutter Instrument’s web site at

www.sutter.com

4.2

4.2 Heating Filament

Heating Filament

4.24.2

Heating FilamentHeating Filament

The type and size of glass that you choose may require a Heating Filament other than the one installed in your puller at the factory. Please refer to the Heating Filament section of this manual to determine the appropriate style and size of filament necessary for pulling the specific glass you would like to use. This instrument is designed to accommodate any of the Sutter Instrument Trough-type or Box-type filaments that are shown in the Sutter Instrument catalogue. This selection of replacement filaments can also be viewed on Sutter Instrument’s web site at

. Any glass with comparable technical specifications can be used with the P-97.

www.sutter.com

ATIN G FILA ME NT SPECI FICAT I ON S

ATIN G FILA ME NT SPECIATIN G FILA ME NT SPECI

.

FICA TI ON S

FICA TI ON SFI CA TI ON S

ONLY USE SUTTER

ONLY USE SUTTER----SUPPLIED REPLACEMENT HEATING F

ONLY USE SUTTERONLY USE SUTTER INSTRUMENT!

INSTRUMENT!

INSTRUMENT!INSTRUMENT!

Systematic instructions for replacing the Heating Filament can be found in the Maintenance section of this manual.

SUPPLIED REPLACEMENT HEATING FILAMENTS IN THIS

SUPPLIED REPLACEMENT HEATING FSUPPLIED REPLACEMENT HEATING F

ILAMENTS IN THIS

ILAMENTS IN THIS ILAMENTS IN THIS

P-97 FLAMING/BROWN MICROPIPETTE PULLER OPERATION MANUAL – REV. 2.03 – DOM (20091301)

9

5.

5. UN PA CKING

UNPA CKING

5.5 .

UNPA CKINGUN PA CKING

Make certain that you have received all of the following items in the P-97 shipping box:



P-97 micropipette puller



Power cord



Sample box containing the following types of glass:

****

 BF100-50-10 (Microinjection)  BF150-86-10 (Thick-Walled Patch (dissocated or cultured cells))  BF150-110-10 (Thin-Walled Patch (slice or whole tissue))



Four spare heating filaments



Warranty registration



Sutter Instrument Product Catalog



Pipette Cookbook

The Model P-97 micropipette puller is shipped to you in a custom box with foam inserts.. Please save shipping materials for future use. Should it ever be necessary to ship the puller to another location, the same packaging should be used to prevent damage to the instrument. Additional packing material may be purchased from Sutter Instrument Company.

IMPORTANT: Improper packaging is a form of abuse and, as such,

IMPORTANT: Improper packaging is a form of abuse and, as such, can be responsible for

IMPORTANT: Improper packaging is a form of abuse and, as such, IMPORTANT: Improper packaging is a form of abuse and, as such, voiding the warranty where shipping damage is sustained because of such packing

voiding the warranty where shipping damage is sustained because of such packing.

voiding the warranty where shipping damage is sustained because of such packingvoiding the warranty where shipping damage is sustained because of such packing

can be responsible for

can be responsible for can be responsible for

*

Unless specified otherwise, the P-97 is shipped equipped with, and configured for, a 2.5mm box heating filament.

P-97 FLAMING/BROWN MICROPIPETTE PULLER OPERATION MANUAL – REV. 2.03 – DOM (20091301)

11

6.

6. SE TT IN G UP

SET T IN G UP

6.6 .

SET T IN G UPSE TT IN G UP

6.1

6.1 Line Power (Mains)

Line Power (Mains)

6.16.1

Line Power (Mains)Line Power (Mains)

The power cord provided with the P-97 connects to the Power Entry Module located on the back of the unit (see diagram below). This Module also includes the Line Fuse.

Figure 6-1. P-97 Cabinet (rear view).

Make certain that the Power Switch located on the left end of the P-97 cabinet is turned OFF.

Figure 6-2. P-97 Cabinet (end view, left).

Plug the power cord provided with the P-97 into the Line Input socket on the Power Entry Module and then to a power source of the appropriate voltage and frequency.

Figure 6-3. Power connection.

P-97 FLAMING/BROWN MICROPIPETTE PULLER OPERATION MANUAL – REV. 2.03 – DOM (20091301)

13

7.

7. FR O NT PA N EL

FRON T PANE L

7.7 .

FRON T PANE LF RO NT PAN EL

Figure 7-1. P-97 Front Panel.

7.1

7.1 Controls

Controls

7.17.1

ControlsControls

Vacuum Fluorescent Display

Vacuum Fluorescent Display Displays program parameters.

Vacuum Fluorescent DisplayVacuum Fluorescent Display

Reset

Reset Re-initializes the controller.

ResetReset

Ke

Keypad

ypad Used to program parameter values and execute

KeKe

ypadypad

programs.

Keys:

Keys:Keys:

0000----9999 Used for choosing the desired program or control

function, entering numeric values when programming and to make YES/NO (1/0) decisions.

CLR

CLR Used to delete programs or numeric values entered into

CLRCLR

those programs. This key is also used to access the RAMP TEST.

ENTR

ENTRENTR

NEXT Used to move to the next line in a program while

NEXTNEXT

editing.

LAST

LAST Used to move to the previous line in a program while

LASTLAST

editing.

PULL

STOP

PULL Initiates the execution of a program.

PULLPULL

STOP Aborts the execution of a program.

STOPSTOP

P-97 FLAMING/BROWN MICROPIPETTE PULLER OPERATION MANUAL – REV. 2.03 – DOM (20091301)

14

7.2

Write Protection (Off)

Last Date/Time edited

Pressure Setting

Program #

Program Line #

7.2 Display

Display

7.27.2

DisplayDisplay

The P-97 has a two line vacuum fluorescent display for easy viewing from any angle. The following figure demonstrates what you will see after you have selected a program. A brief description of each parameter is provided below.

UNLOCKED P=500 JAN 18, 1997 2:30 PM

UNLOCKED P=500 JAN 18, 1997 2:30 PMUNLOCKED P=500 JAN 18, 1997 2:30 PM 3 1 HEAT=330,PULL=150,VEL.= 80,TIME=200

3 1 HEAT=330,PULL=150,VEL.= 80,TIME=200

3 1 HEAT=330,PULL=150,VEL.= 80,TIME=2003 1 HEAT=330,PULL=150,VEL.= 80,TIME=200

Figure 7-2. Program Display.

PROGRAM

PROGRAMPROGRAM (0-99)

A program consists of one or more cycles when executed in sequence, will ‘pull’ the capillary

1111

1

which,

glass inserted in the instrument. A program can be up to 8 cycles in length

WRITE PROTECTION

WRITE PROTECTIONWRITE PROTECTION (Locked/Unlocked)

PRESSURE SETTING

PRESSURE SETTINGPRESSURE SETTING (Range 0-999)

DATE/TIME

DATE/TIME Reports the date and time of the last program edit.

DATE/TIMEDATE/TIME

HEAT

HEATHEAT (Range 0-999)

"

locked/unlocked"

}}}}Prevents program from being

edited when in Locked mode.

Reports the programmed value of the air pressure during the active cooling phase of the pull cycle.

HEAT controls the level of electrical current supplied to the filament. The HEAT required to melt a piece of glass is a function of the filament installed and the particular glass size and composition. It is important that the HEAT value be set relative to the Ramp Test value as discussed in the Operation Section. Generally, changes to HEAT will be made in steps of about 5 units since in most cases smaller changes will have little effect.

PULL

PULLPULL (Range 0-255)

1

A CYCLE consists of four programmable parameters; H

sequence of events during execution of a cycle is described in the PROGRAMS SECTION of this manual.

P-97 FLAMING/BROWN MICROPIPETTE PULLER OPERATION MANUAL – REV. 2.03 – DOM (20091301)

This parameter controls the force of the hard pull. In general, the higher the PULL, the smaller the pipette’s tip diameter and the longer the taper. Useful changes in PULL strength are 10 units or more to see an effect.

EAT

, P

ULL

, V

ELOCITY

and either T

IME or DELAY

. A CYCLE is equivalent to one line of program code. The

15

VELOCITY

VELOCITYVELOCITY (Range 0-255)

TIME

TIMETIME (Range 0-255)

The velocity of the glass carriage system is measured as the glass softens and begins to pull apart under a constant load. The increasing velocity of the initial pull is determined by the viscosity of the glass, which in turn is a function of the glass temperature. The adjustable velocity allows for a selection of a precise glass temperature as the trip point for the hard pull. Useful values for velocity range from 10 to 150 with lower values being used for patch and injection pipettes and higher values for micropipettes. SEE THE PROGRAMS SECTION FOR A DISCUSSION OF THE SIGNIFICANCE OF VELOCITY=0.

TIME is one of two available modes of cooling and controls the length of time the cooling air is active. If VEL>0 then one unit of TIME represents 1/2ms. If VEL=0 then one unit of TIME represents 10ms. SEE THE PROGRAMS SECTION FOR A DISCUSSION OF THE SIGNIFICANCE OF TIME=0.

DELAY

DELAYDELAY (Range 0-255)

DELAY is a cooling mode, which controls the delay time between when the heat turns off and when the hard pull is activated. The gas is automatically turned on for 300ms. The higher the DELAY value, the cooler the glass will be when the hard pull is executed. Thus, increasing the DELAY results with decreased taper length and increased tip diameter. If VEL>0 then one unit of DELAY represents 1/2ms. If VEL=0 then one unit represents 10ms. SEE THE PROGRAMS SECTION FOR A DISCUSSION OF THE SIGNIFICANCE OF DELAY=0.

P-97 FLAMING/BROWN MICROPIPETTE PULLER OPERATION MANUAL – REV. 2.03 – DOM (20091301)

17

8.

8. ME CH AN ICAL D ES CRIPTI

MEC H AN ICAL D ES CRIPTI ON (P U LLER A NA TOMY)

8.8 .

MEC H AN ICAL D ES CRIPTIMEC H AN ICAL D ES CRIPTI

8.1

8.1 Some Basic Information

Some Basic Information

8.18.1

Some Basic InformationSome Basic Information

This section presents a basic mechanical description of the P-97, with particular emphasis on terminology. Knowing the names of the various parts greatly facilitates communication between the investigators and the manufacturer when discussing adjustments or service problems. In addition, various controls and adjustments on the top of the instrument are located and described. Those adjustments, which are considered part of maintenance procedures, are dealt with in the Maintenance Section of this manual.

8.2

8.2 Air Cooling System

Air Cooling System

8.28.2

Air Cooling SystemAir Cooling System

The Model P-97 supplies a blast of air to cool the filament area after the heating segment of a pull cycle. The components of the air-cooling system are shown below.

ON ( PU LLER A NA TOMY)

ON ( PU LLER A NA TOMY)ON ( PU LLER A NA TOMY)

Figure 8-1. P-97 Base Plate (Top view, cover removed).

Air Compressor

Air Compressor The air compressor (or pump) creates the air pressure used

Air CompressorAir Compressor

to cool the filament and glass during the pull cycle.

Air Jet Nozzle

Air Jet Nozzle Directs the cooling air to the filament. The air jet nozzle

Air Jet NozzleAir Jet Nozzle

should be positioned 2 to 3 millimeters below the filament. The screw that secures the air jet to the filament block can be loosened allowing the jet to move up and down.

Air Valve Solenoid

Air Valve Solenoid Regulates the flow of cooling air to the filament and glass.

Air Valve Solenoid Air Valve Solenoid

Drierite Canister

Drierite CanisterDrierite Canister (Reservoir Tank)

(Reservoir Tank)

(Reservoir Tank)(Reservoir Tank)

A canister filled with Drierite used to remove moisture from the air flowing between the pump and the valve solenoid. The dissected air is used to purge heat before and after a pull and to cool the air flowing to the filament and glass during the pull cycle.

P-97 FLAMING/BROWN MICROPIPETTE PULLER OPERATION MANUAL – REV. 2.03 – DOM (20091301)

SUTTER INSTRUMENT P-97 Operation Manual

Specifications and Main Features

Frequently Asked Questions

User Manual