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P-97 FLAMING/BROWN MICROPIPETTE PULLER OPERATION MANUAL – REV. 2.43 - DOM (20161118)

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Summary of Contents for P-97

Page 1: ...P 97 Flaming Brown Micropipette Puller Operation Manual Rev 2 43 DOM 20161118 One Digital Drive Novato CA 94949 Voice 415 883 0128 Web www sutter com Fax 415 883 0572 Email info sutter com...

Page 2: ...P 97 FLAMING BROWN MICROPIPETTE PULLER OPERATION MANUAL REV 2 43 DOM 20161118 ii Q Copyright 2016 Sutter Instrument Company All Rights Reserved Flaming Brown is a trademark of Sutter Instrument...

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Page 5: ...voltage This instrument is designed for connection to a standard laboratory power outlet Overvoltage Category II and because it is a microprocessor controlled device it should be accorded the same sy...

Page 6: ...relative humidity at 40 C 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 sin...

Page 7: ...ooling System 5 1 5 3 Heating Assembly 5 1 5 4 Upper Pulley Assembly 7 1 6 Cabinet 8 1 6 1 Electronics 8 2 INSTALLATION 9 2 1 Unpacking 9 2 2 Setting Up 9 2 2 1 Line Power Mains 9 3 OPERATING INSTRUCT...

Page 8: ...oint 25 3 5 3 4 Cooling 25 3 5 3 5 PRESSURE 25 3 5 4 Step by Step Patch Pipette Programming 25 3 5 5 Technical Tips for Pulling Micropipettes 27 3 5 5 1 Regulating the Time it takes to pull a Sharp Pi...

Page 9: ...e made to pull straight 40 5 1 8 Problem One electrode is much longer than the other electrode 40 5 1 9 Problem The shape and resistance of the pipette changes from pull to pull 41 5 2 Controller prob...

Page 10: ...polishing spacer 28 Figure 3 11 Fire Polish Spacer in Puller Bar 29 Figure 3 12 Filament alignment 29 Figure 3 13 End view of box filament and glass 30 Figure 3 14 Box filament positioning 31 Figure...

Page 11: ...sses 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...

Page 12: ...is 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...

Page 13: ...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 pa...

Page 14: ...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 th...

Page 15: ...sor or pump creates the air pressure used to cool the filament and glass during the pull cycle Air Jet Directs the cooling air to the filament The air jet should be positioned 2 to 3 millimeters below...

Page 16: ...down so that the filament is positioned at the correct level relative to the glass If the jaws are repositioned and or the filament type has been changed make sure that the securing screws of the jaw...

Page 17: ...nstrument Technical Support Figure 1 5 Upper cable pulley assembly Panels Left And Right K in Figure 1 5 The panels are the angled surfaces that provide mountings for the Puller Bars and their Bearing...

Page 18: ...The cable is terminated with crimped on clamps or swages at the back end of each Puller Bar Glass Stop Mounting U in Figure 1 5 Holes for mounting the glass stop Either the left side or the right may...

Page 19: ...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 packin...

Page 20: ...ake certain that the Power Switch located on the left end of the P 97 cabinet is turned OFF Figure 2 2 P 97 Cabinet end view left showing power switch Plug the power cord provided with the P 97 into t...

Page 21: ...r 3 Turn Power switch on left side of cabinet ON 4 Press 0 followed by ENTR on the keypad to view Program 0 The cursor will be flashing on the HEAT parameter 5 Inspect the parameter values displayed f...

Page 22: ...rogram Structure The resulting size and shape of a micropipette made using the P 97 is determined by the parameter values that are programmed by the user One hundred 100 separate programs can be saved...

Page 23: ...ammed 1 unit equals 10ms This feature allows you to use the puller to fire polish the resulting patch pipette TIME Range 0 255 Controls the length of time the cooling air is active This parameter is o...

Page 24: ...e enclosed program sheet 0 Micropipette tip diameter less than 0 1 m 1 Patch pipette In describing the operation of the puller the above configuration is assumed If the configuration of your puller di...

Page 25: ...o scroll to previous lines Press the LAST key 3 2 7 Clearing a Program from Memory CLR To clear the values of a particular program from memory you can utilize the CLEAR function Pressing the CLR key g...

Page 26: ...to the VELOCITY field Note that the values for HEAT and PULL remain unchanged Enter a new value e g 90 if the value is two digits you must press ENTR to enter the value The cursor will be positioned...

Page 27: ...ish or adjust program HEAT values as a function of the filament glass combination This test should be run when using the puller for the first time before writing or editing a program whenever you chan...

Page 28: ...this function is accessed Changes of less than 10 units will not be noticeable Air pressure is reported on the program header line Range 1 to 999 Default 500 Recommendations For thick walled glass 500...

Page 29: ...ull and the length of time the end of pull information is displayed Units are in seconds The time you enter is actually an offset added to an automatic 3 second chamber cooling cycle e g A setting of...

Page 30: ...amiliar with the pulling process 1 Load the glass into the puller as described previously in the FIRST TIME USE chapter 2 Press 0 followed by ENTR on the keypad to view Program 0 3 Inspect the paramet...

Page 31: ...n There is always the possibility that the puller will be given a set of values that stall its operation For example the HEAT value has not been set high enough to melt the glass thus the glass cannot...

Page 32: ...on pipettes select a HEAT value that melts the glass in about 7 seconds or longer 3 5 2 2 PULL Strength Low values of PULL strength in the range of 50 to 75 will give larger tips appropriate for injec...

Page 33: ...g be used The method for changing to the DELAY mode is described under Software Control Functions and a brief description of the two modes of cooling is given in Programs After switching to DELAY mode...

Page 34: ...ys The trip velocity is lower Hard pull is not activated PULL 0 More than one heating cycle is used The P 97 can be used very effectively for this type of processing The following general information...

Page 35: ...and or needing short tapered pipettes The DELAY mode provides a set 300 milliseconds of cooling 3 5 3 5 PRESSURE The recommended pressure setting when using thick walled glass is 500 or greater For t...

Page 36: ...rogram just established may be sufficient for your application However changes made in a one line program are amplified throughout the cycle potentially producing gross changes in the pipette If you n...

Page 37: ...ramp value plus 5 electrode length is controlled by air pressure adjustments If this is insufficient a wider or more narrow filament can be installed 3 5 5 2 1 Filament Width Filaments narrower than...

Page 38: ...filament and use the Fire Polish Spacer block described next Using the adjusting screw on the top of the spacer position the tip of the pipette just inside the edge of the filament 4 Press Pull and t...

Page 39: ...ch filament to use for your application contact our technical support staff 3 6 2 Filament Positioning and Aligning Figure 3 12 Filament alignment Adjust the filament position using the two eccentric...

Page 40: ...lowing micropipettes Patch pipettes using thick walled glass Microelectrodes used for slice preparations where long parallel walls would aid penetration Microinjection needles for transgenic research...

Page 41: ...2mm square x 3 0mm wide 1 0mm FB255B Standard 2 5mm square x 2 5mm wide 2 0mm FB245B 2 5mm square x 4 5mm wide 2 0mm FB315B 3mm square x 1 5mm wide 1 5mm FB320B 3mm square x 2 0mm wide 1 5mm FB330B 3...

Page 42: ...he distance between the top corners distance A is approximately 2 3 the length of the bottom of the filament This geometry will provide improved heat distribution to the top of the glass tubing When r...

Page 43: ...at the lower left corner of the Plexiglas cover plate and pull the chamber straight off 2 Loosen the two clamp screws D in Figure 4 2 only one shown that hold the filament in place and then slide out...

Page 44: ...ter the glass you will need to reposition the upper and lower heater jaw assemblies by first loosening the brass screws holding the jaws to the black nylon D Reposition the jaws then retighten the bra...

Page 45: ...ity and or a disparity between the taper lengths of the pair of pulled pipettes as illustrated in Figure 4 4 Taper length inequality is generally caused by the air jet not being aimed at the center of...

Page 46: ...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...

Page 47: ...te with the cap to the left and the air tube connections to the front The newer style plastic hold downs can be tightened by reinstalling the front screws The older style black plastic hold downs slid...

Page 48: ...P 97 FLAMING BROWN MICROPIPETTE PULLER OPERATION MANUAL REV 2 43 DOM 20161118 38 This page intentionally blank...

Page 49: ...e length of the electrode and what is used for the filling solution If the same solution is used then resistance may give an indication of how well the electrode will penetrate a cell as the electrode...

Page 50: ...mm tip that also has a very short final taper and tip 20 to 50 mm long How can this be done See Bee Stinger pipettes in Chapter 3 of the Pipette Cookbook Try a program in which the first two lines of...

Page 51: ...ment it is quite possible that the new filament needs a very different HEAT value than what you have been using It is always a good idea to run the ramp test each time you change the filament 3 If you...

Page 52: ...al Support 5 2 3 Problem Display only shows a cursor 1 The microprocessor has failed to properly initialize the display Press RESET and the display should show the proper power up message Always allow...

Page 53: ...by Sutter Instrument during the first three months this warranty is in effect after which the cost is the responsibility of the customer The limited warranty is as stated above and no implied or infer...

Page 54: ...P 97 FLAMING BROWN MICROPIPETTE PULLER OPERATION MANUAL REV 2 43 DOM 20161118 44 This page intentionally blank...

Page 55: ...ase contact Sutter Instrument for more information FS1875 Platinum iridium sheet 18 x 75 x 0 05 mm 0 002in Capillary Glass Tubing Several sizes and types of borosilicate and aluminosilicate glass tube...

Page 56: ...P 97 FLAMING BROWN MICROPIPETTE PULLER OPERATION MANUAL REV 2 43 DOM 20161118 46 This page intentionally blank...

Page 57: ...he fuse that is readily visible in the fuse holder when you take it out is the one that is active when the holder is installed A spare fuse is also stored within the fuse holder It is concealed in a c...

Page 58: ...L REV 2 43 DOM 20161118 48 Figure 5 2 Fuse holder Replace fuse only with the same type and rating Type Medium Time Delay or Time Lag 5 x 20 mm glass tube RoHS compliant Rating 3A 250V Examples Bussman...

Page 59: ...Delay IEC 60127 2 Sheet III or Time Lag Rating 3A 250V Examples Bussmann GMC 3A or GMC 3 R or Littelfuse 239 003 or 239 003 P Internal safety fuses not operator accessible Red line Type T slow blow 0...

Page 60: ...P 97 FLAMING BROWN MICROPIPETTE PULLER OPERATION MANUAL REV 2 43 DOM 20161118 50 This page intentionally blank...

Page 61: ...AMS PER LITER MELTING POINT 1450 C DECOMPOSES APPEARANCE BLUE GRANULES NO ODOR SECTION IV FIRE AND EXPLOSION HAZARD DATA FLASH POINT NONE EXTINGUISHING MEDIA NOT COMBUSTIBLE SPECIAL FIREFIGHTING PROCE...

Page 62: ...RATING DUST SECTION VIII CONTROL MEASURES RESPIRATORY PROTECTION NIOSH OSHA APPROVED FOR DUST VENTILATION TO MEET TLV REQUIREMENTS EYES SAFETY GLASSES OR GOGGLES OTHER PROTECTIVE EQUIPMENT GLOVES OR P...

Page 63: ...test 17 controls front panel 3 cycle definition 3 12 D date 4 default configuration 14 20 delay definition 4 13 desiccant regeneration 36 dimensions 49 of glass 1 disclaimer iii display 3 vacuum fluo...

Page 64: ...22 microinjection pipettes 13 microperfusion pipettes 13 micropipette fabrication 22 N notes user 56 O OD 1 operation 20 first time use 11 precautions iv P P80 PC programs 19 P 87 programs 18 panels...

Page 65: ...Canister 36 Replacing the Granules 36 reset 3 S safety warnings iii 48 electrical iii mains fuse iii safety warnings precautions iii solenoid 5 spring stops 7 T technical specifications 49 technical s...

Page 66: ...56 NOTES P 97 FLAMING BROWN MICROPIPETTE PULLER OPERATION MANUAL REV 2 43 DOM 20161118...

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