Related Manuals for Sutter Instrument P-97
Summary of Contents for Sutter Instrument P-97
- Page 1 OPERATION MANUAL P-97 Flaming/Brown Micropipette Puller rev. 0903 - DOM SUTTER INSTRUMENT COMPANY...
- Page 2 P-97 Flaming/Brown Micropipette Puller Operation Manual (rev. 0903) DOM Manufactured by Sutter Instrument Company 51 Digital Drive Novato, CA 94949 U.S.A. voice:415-883-0128 web: http://www.sutter.com fax:415-883-0572 email: info@sutter.com...
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Page 3: Table Of Contents
TABLE OF CONTENTS GENERAL INFORMATION..............................1 ..................................1 ECHNICAL UPPORT ’ P-97? ................................1 S NEW ON THE SAFETY WARNING..................................3 PRECAUTIONS .................................... 4 ....................................4 N OPERATION ................................ 4 N HANDLING MICROPIPETTES GLASSWARE & HEATING FILAMENT SPECIFICATIONS ..................5 UNPACKING .................................... - Page 4 PULLING PIPETTES ................................27 ....................................27 ROCEDURES ..............................28 OTES ON ROGRAM PERATION PARAMETER ADJUSTMENT ..............................29 ................................29 ENERAL NFORMATION ......................29 ICROPIPETTE ICROINJECTION EEDLE ABRICATION ...............................31 ATCH IPETTE ABRICATION ..........................35 ECHNICAL TIPS FOR PULLING MICROPIPETTES ....................................36 OLISHING HEATING FILAMENTS ................................38 ................................38 ENERAL NFORMATION ..................................38 ROUGH ILAMENT .....................................39...
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Page 5: General Information
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. - Page 6 · Write protection to prevent programs from being corrupted · Displayed date/time of last program edit · Increased memory capable of storing 100 programs · Switchable cooling mode to facilitate easy transfer of programs from both P-80PC and P- 87 pullers...
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Page 7: Safety Warning
· To avoid injuring your back or limbs it is recommended that you do not attempt to lift this instrument by yourself. The P-97 Mi- cropipette Puller weighs in excess of 18kg and should be moved by TWO people. -
Page 8: Precautions
The P-97 is designed to be operated in a laboratory environment (pollution · degree II) and at temperatures between 5°C - 40°C. The P-97 is designed to be connected to a standard laboratory power outlet · (overvoltage category II) with main supply voltage fluctuations not to exceed ±... -
Page 9: Glass Capillary & Heating Filament Specifications
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 Company’s World Wide Web site at www.sutter.com. -
Page 10: Unpacking
· Catalog The Model P-97 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. -
Page 11: Setting Up
Input Figure 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 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. -
Page 12: First Time Use
FIRST TIME USE While we realize that most new users of the P-97 are anxious to start pulling useable pipettes right away, we cannot over-state the importance of taking a few moments to review the manual in order to understand how the puller works. Many a heating filament has been destroyed with first use because the user did not understand the relationship between the programmable heat settings and the filament installed in the puller. - Page 13 Press the < PULL > key on the keypad. The heating filament should turn on and the glass should separate in less than 10 seconds. Loosen the clamping knobs and remove the pipettes from the puller bars. Unless otherwise stated on your program sheet, Program 0 is factory pre-programmed to pull a micropipette (tip diameter less than 0.1 mm) from 1.0mm x 0.5mm borosilicate glass.
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Page 14: Front Panel
FRONT PANEL Figure 5. P-97 Front Panel Controls Vacuum Fluorescent Display Displays program parameters Reset Re-initializes the controller Keypad Used to program parameter values and execute pro- grams Used for choosing the desired program or control func- tion, entering numeric values when programming and to... -
Page 15: Display
Display 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. Pressure Setting... - Page 16 PULL This parameter controls the force of the hard (Range 0-255) pull. In general, the higher the PULL, the smaller the pipette tip diameter and the longer the taper. Useful changes in PULL strength are 10 units or more to see an effect. VELOCITY The velocity of the glass carriage system is (Range 0-255)
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Page 17: Mechanical Description (Puller Anatomy)
Maintenance Section of this manual. Air 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. -
Page 18: Heating Assembly
Heating Assembly The HEATING ASSEMBLY comprises the FILAMENT, FILAMENT BLOCK ASSEMBLY and the CHAMBER. The FILAMENT BLOCK ASSEMBLY and the CHAMBER are discussed below. FILAMENTS are discussed in a separate section. CHAMBER (Figure 8) The chamber encloses the filament block assembly to provide a controlled environment in which to pull the glass. -
Page 19: Upper Cable Pulley Assembly
COVER PLATE (Figure 8) The cover plate conceals the entry of the PULLING CABLES into the BASE of the instrument. It is attached to the top by two screws, in slots, at points labeled C. Loosening these screws allows the FILAMENT BLOCK/ ANGLE PLATE assembly to move forward and back over large distances. -
Page 20: Cabinet
Electronics The P-97 micropipette puller is controlled by a Z-80 microprocessor. Three digital-to-analog (D-A) converters control the HEAT, PULL and VELOCITY values. The HEAT power supply is a precision constant current switching unit which will vary less than 10 milliamperes with a plus or minus 10% change in the a.c. -
Page 21: Programs
PROGRAMS Program structure The ultimate size and shape of a micropipette made using the P-97 is determined by the parameter values that are programmed by the user. 100 separate programs can be saved for future use. Each program is structured as follows:... - Page 22 VELOCITY The VELOCITY of the glass carriage system is measured (Range 0-255) 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.
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Page 23: Pull Cycle
Pull cycle A typical PULL CYCLE in a PROGRAM is described below: Figure 10. Pull Cycle when TIME > 0 The heat turns on. The glass heats up and a weak pull draws the glass out until it reaches the programmed velocity. -
Page 24: Default Configuration
COPYRIGHT SUTTER INSTRUMENT CORP. 1992 Figure 12. Power On Display The P-97 has the capacity to store 100 programs (0 - 99). On the keypad, press < 0 > followed by < ENTR > (factory installed micropipette program) or the number of another program you wish to execute. -
Page 25: Viewing A Program
3 1 HEAT=330, PULL=150, VEL.=80, TIME=200 Figure 13. P-97 Program Display Press < RESET > to bring up the sign-on prompt To select a different program, then press the number of the desired program (0 to 99) followed by < ENTR >. -
Page 26: Editing A Program
Editing a program Select a program number (e.g. 10). If there are already values entered, make sure that this program was not entered by another user of the puller. Unused program areas are usually cleared before a puller is shipped, but occasionally test program values are inadvertently left in memory. -
Page 27: Software Control Functions
DO YOU WISH TO CLEAR ALL VALUES FROM THE PRESENT CYCLE TO THE END? NO=0 YES=1 Figure 15. Access to Control Functions Entering < 0 > will provide you with access to the following P-97 CONTROL FUNCTIONS: Function Function Name... -
Page 28: Ramp Test < 1
RAMP TEST < 1 > The HEAT value required to melt your glass is based on the characteristics of the heating filament that is installed and the glass you are using. The RAMP TEST allows you to systematically establish or adjust program HEAT values as a function of the filament/glass combination. -
Page 29: Change Air Pressure < 2
TIME MODE Recommendations Use the TIME MODE for: patch pipette programs glass <1.2mm O.D. transfer of model P-87 puller programs to P-97 Use the DELAY MODE for: single cycle programs glass >1.2mm O.D. transfer of model P80-PC puller programs to P-97. -
Page 30: Air Time At Start Of Pull < 4
AIR TIME AT START OF PULL < 4 > Controls the length of time the humidity control chamber is purged with dehumidified air prior to execution of the pull sequence. The intent of the control is to minimize and stabilize the chamber humidity during the pull sequence. -
Page 31: Pulling Pipettes
Parameter Adjustment section of this manual. A unique feature of the P-97 is its capability to loop through a program. This is demonstrated with the multicycle Program 1, which is factory pre-programmed to pull a patch type pipette with a tip diameter of about 2mm. -
Page 32: Notes On Program Operation
UNLOCKED, P = 500, JAN 18 1993 2:35 PM 1 1 HEAT = 285, PULL = 0, VEL. = 50 TIME = 150 Figure 17. Sample Program Adjust the HEAT in the program to Ramp value plus 15 units. Load glass into the puller and press <... -
Page 33: Parameter Adjustment
PARAMETER ADJUSTMENT General Information Micropipette and microinjection needle programs are sufficiently different from patch pipette programs that the following information on parameter adjustments has been divided into three sections: Micropipette/microinjection needle fabrication, Patch pipette fabri- cation and Technical Tips. Even if your research only requires one type of pipette, we recommend that you read through all three sections for full appreciation of the capabilities of the puller. -
Page 34: Velocity (Trip Point)
VELOCITY (trip point) The VELOCITY value determines the point at which the heat is turned off. VELOCITY reflects the speed at which the two carrier bars are moving during the weak pull. The lower the VELOCITY value the slower the speed of the bars when the trip point occurs. Although VELOCITY can safely be set to any value from 1-255, all values over a maximal trip point (about 150) will produce equivalent results. -
Page 35: Pressure Adjustment
With a range of control over the degree of pipette cooling one can control of the rapidity of pipette taper. Longer DELAY values increase cooling and form a pipette with a more rapid taper (shorter shank) while shorter DELAY values decrease cooling and form a pipette with a more gradual taper (longer shank). -
Page 36: Patch Pipette Fabrication
The PULL strength is typically turned off. More than one heating cycle is used. The P-97 can be used very effectively for this type of processing. The following general information will familiarize you with the effect of adjusting each of the pull cycle parameters in a typical patch pipette program. - Page 37 TIME For patch pipette programs, it is advisable to keep the TIME between 200 and 250 to maxi- mize the cooling of the glass. DELAY mode of active cooling is not used in patch pipette fabrication. By contrast to micropipette pulling (see above), the TIME settings in the upper range of available values can have a pronounced effect on glass cooling.
- Page 38 Repeat step (3) only this time decrease the VELOCITY. As the VELOCITY decreases, the number of loops increases. By adjusting the VELOCITY as described, establish the number of loops required to approximately form a pipette with the characteristics you desire. Set the VELOCITY value in your program to the number that falls midway between the values required to loop one more and one less times than the desired number.
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Page 39: Technical Tips For Pulling Micropipettes
Technical tips for pulling micropipettes Regulating the time it takes to pull a sharp pipette HEAT. For 1.0mm O.D. tubing, if the pull takes longer than eight seconds and you are trying to pull a fine micropipette, increase the HEAT. To do this in a methodical fashion, increase the HEAT value in five unit increments, each time monitoring pull time until the pull takes place in less than eight seconds. -
Page 40: Fire Polishing
Fire Polishing The P-97 micropipette puller allows you to perform pseudo fire-polishing of pipette tips but does not have a provision for visualizing the pipette tip during the heating process. The extent of the heating required to attain the desired degree of polishing must be empirically established. - Page 41 Figure 20: Fire Polish Spacer in Puller Bar...
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Page 42: Heating Filaments
HEATING FILAMENTS General Information The pipette programs that you ultimately develop will largely depend on the type of heating filament installed in the puller and the glass that you use. Depending on your research application, there may be an optimum filament/glass combination which differs from the configuration the puller is currently set up with. -
Page 43: Box Filament
The trough filament you select depends upon the length of the taper that you want. Wider filaments for special purposes are available upon request. Filament Description FT315B 1.5mm wide trough FT320B 2mm wide trough FT330B 3mm wide trough FT345B 4.5mm wide trough Table 3. - Page 44 The box filament has two primary limitations. First, it requires more current to heat to a given temperature than the same size trough filament. Thus it is possible to use wider trough filaments without exceeding the maxi- mum current capacity of the puller. Second, the box configuration reduces the cooling effect of the air jet.
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Page 45: Maintenance
MAINTENANCE Cleaning To maintain the P-97 in optimal condition the vinyl dust cover that is shipped with the P-97 should be used whenever the unit is turned off to protect the puller from dust and spills. Occasionally clean the exterior and the base plate of the unit by wiping them with a dry cloth to remove dust and fine pieces of glass. - Page 46 Identify the flathead locking screws to the right of each chrome screw. Figure 24. Filament Alignment Loosen the locking screws (A’ and B’) Turn the eccentric chrome screw (A) on the vertical face to adjust the vertical position of the filament and the eccentric chrome screw (B) on the horizontal face to adjust the front-to-back position of the filament.
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Page 47: Pulley Adjustment
Figure 25. Micropipette Shapes Pulley Adjustment The position of the two pulleys (F in Figure 26A) which carry the cables from the solenoid (not shown) to the carriers (G in Figure 26A) is adjustable. The pulley position controls the tension of the cables. A difference between the tension of the two cables can cause problems with pipette reproducibility and/or a disparity between the taper length of the pair of pulled pipettes (as illustrated in Figure 25). -
Page 48: Regeneration Of Drierite Granules
The Indicating Drierite found in the canister at the right rear corner of the base plate on the P-97 is a desiccant made of calcium sulfate (97%) and cobalt chloride (3%). This material is used to remove water vapor from the air-cooling supply system. As it absorbs more moisture it becomes pink in color and must eventually be “regenerated”... - Page 49 then be cooled in a tight container before refilling the plastic canister. Filters should also be ° pre-dried at 100 C for about 30 minutes before assembly. Drierite is not toxic and can be handled with few precautions. For more detailed safety information please refer to the enclosed MATERIAL SAFETY DATA SHEET.
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Page 50: Troubleshooting
If however the HEAT is such that the pull takes place in more then 8 seconds, decreasing the cooling may somewhat decrease the tip size. Cooling can be most effectively decreased in the P-97 by decreasing air pressure, however a decreasing TIME may also be useful. - Page 51 Problem: HOW DO I INCREASE THE SIZE OF MY PATCH-PIPETTE? The first thing to try is to reduce the HEAT. Try dropping the HEAT 5 units at a time to see if this will increase the size of the tips. If this does not work increase the pressure in units of 50.
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Page 52: Problem: One Electrode Is Much Longer Than The Other Electrode
Problem: ONE ELECTRODE IS MUCH LONGER THAN THE OTHER ELECTRODE. This is caused by one of two things. The filament may not be centered over the air jet. If it is not, follow the procedure “Testing the position” in the Heating filament replacement section of the Mainte- nance chapter. -
Page 53: Problem: Display Blank, Fan Not On
If the values entered are not held when the power is turned off, a failure in components that are not serviceable by the user has likely occurred. Contact Sutter Instrument Company Technical Support. -
Page 54: Fuse Replacement
Replace the active fuse with the spare and re-install the fuse holder and power cord. If the controller fails to power up with the new fuse installed, call Sutter Instrument technical support personnel for assistance. pry here to remove... -
Page 55: Limited Warranty
LIMITED WARRANTY Sutter Instrument Company, a division of Sutter Instrument Corporation, limits the warranty on this instrument to repair or replacement of defective components for one year after the date of shipment, provided the instrument has been operated in accordance with the instruc- tions outlined in the instruction manual. -
Page 56: Disclaimer
DISCLAIMER The pipette puller Model P-97 is designed for the specific use of creating micropipettes and no other use is recommended. This instrument creates items which should only be used in a laboratory environment for use on animal tissues. It is not intended to be used and should not be used in human experimen- tation or applied to humans in any way. -
Page 57: Technical Specifications
TECHNICAL SPECIFICATIONS Dimensions (H x W x D): 12” x 21” x 14” 30cm x 53cm 36cm Weight: 60 lb. 28 kg Electrical: Mains voltage 115 V, 60 Hz Maximum power consumption 370 VA Mains fuse (rear of cabinet) at 115V Type T (slow blow), 3A, 250V, GMC Internal safety fuses (not operator accessible) -
Page 58: Appendix: Adrierite Material Safety Data Sheet
APPENDIX: A DRIERITE MATERIAL SAFETY DATA SHEET IDENTITY: INDICATING DRIERITE DATE PREPARED 1-3-96 DESCRIPTION: 1/16" TO 1/4" BLUE GRANULES SECTION I MANUFACTURER’S NAME: W.A. HAMMOND DRIERITE CO. LTD. ADDRESS: P.0. BOX 460, 138 DAYTON AVE., XENIA, OH 45385 EMERGENCY PHONE NUMBER: (513) 376-2927 INFORMATION PHONE NUMBER: (513) 376-2927 SECTION II... -
Page 59: Section Vii Spill Or Leak Procedures
SIGNS AND SYMPTOMS OF OVER EXPOSURE: EYE, NOSE, THROAT, OR RESPIRATORY IRRITATION CARCINOGENICITY OF INGREDIENTS: MATERIAL IARC OSHA CALCIUM SULFATE NOT LISTED NOT LISTED NOT LISTED COBALT CHLORIDE YES* *(COBALT & COBALT COMPOUNDS ARE CLASSIFIED AS GROUP 2B) MEDICAL CONDITIONS GENERALLY AGGRAVATED BY EXPOSURE: PRE-EXISTING UPPER RESPIRATORY AND LUNG DISEASE SUCH AS, BUT NOT LIMITED TO, BRONCHITIS, EMPHYSEMA &... -
Page 60: Index
INDEX date ................11 compressor..............13 reset (time/date)............26 air jet ..............13,41 default configuration ........20,24-26 air jet adjustment/alignment ....13,38,39,41 delay reservoir tank (drierite cannister) ......13 definition ............... 12,18 solenoid (reedex valve) ..........13 special condition ............18 air mode ..............25 dimensions (puller/instrument) .......53 delay ............ - Page 61 heat definition .............11,17 P80-PC programs .............25 heat value optimal setting ........8,24 P-87 programs............25 heater jaws .............14,41,42 panels, L/R...............15 heating assembly............14 parameter adjustment..........29 angle plate ............. 14,42 Patch pipette fabrication........31-34 chamber............14,23,26 patch pipettes.........9,17,28,31-34 cover plate ............. 14,15 power consumption..........53 filament block............14,42 power cord mains.............53 heater jaws ............
- Page 62 unlock.............11,23,26 unpacking..............6 V-bearings .............15,16,41 velocity ............28,30,32-34 definition ............... 12,17 special condition ............17 voltage mains............4,53 warranty..............51 weight ..............53 write protection..........11,23,26...
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