NVI-570V Automatic Injector User`s Manual
Contents
1. FILL button At this point unplug the power supply connector from the control box count to five and reconnect the power supply An audible beep will be heard The unit is now in the home position and ready to have the backfilled micropipette positioned onto the wire plunger Do not forget to loosen the collet before trying to install a micropipette onto the injector If you cannot see the wire plunger when looking into the collet it is retracted too far into the injector head To reposition press and hold the EMPTY button until the wire plunger tip is visible and flush with the end of the collet Next unplug the power connector from the control box count to five and reconnect An audible beep will be heard and the unit is again in the home position SUGGESTION When you are finished using the unit for the day make sure the wire plunger is in the proper position before it is unplugged B Backfilling the Micropipette For proper injection the pulled micropipettes must be backfilled with oil Inject lightweight mineral oil into the micropipette from the back end until filled Carefully slide the micropipette onto the wire plunger making sure it goes through the first O ring and seats properly into the white spacer This is done by feel and may take some practice Tighten the collet securely At this point push and hold the EMPTY button on the control box The wire plunger will extend forcing exce2. appar ently acts as a brake allowing better control of injection rate 8 An excellent reference to all procedures involving the Xenopus oocyte is Kay B and H Peng Xenopus Laevis Practical Uses in Cell and Molecular Biology Methods in Cell Biology Vol 36 Academic Press CLEANING AND SERVICING INSTRUCTIONS Plastic Sleeve spacer This spacer should be placed between the two black O rings Black O ring large hole Aluminium Barrel i i f a Glass Sealing Black O ring Collet small hole Plunger Note Plastic sleeve spacer should be positioned with shelf facing pipet tip Collet a y E r i Aluminum Barrel Delrin Motor Housing Body Tube DO NOT REMOVE THE MOTOR HOUSING FROM THE DELRIN BODY TUBE DAMAGE TO THE MOTOR WIRES MAY RESULT THIS WILL VOID ALL WARRANTIES Cleaning Recommendations A The Injector see figures 1 and 2 above Remove collet O rings and the spacer Clean these parts and the injector head unit with alcohol Do not soak in liquid or autoclave the NVI 570V A simple wipe down will be sufficient B Control Box Wipe off the box with a clean dry cloth Do not use solvent based cleaners Solvents can dissolve the print on the label and discolor the box Servicing the Unit A Changing the Wire Plunger The most common service function that the end user will perform will be to change t
3. HARVARD APPARATUS Operating Instructions NVI 570V Automatic Injector Catalog No 65 0033 Harvard Apparatus I October Hill Road Ho sachusetts 01746 nc e 84 lliston Mas Toll Free 800 232 2380 Fax 508 429 5732 1 PACKING LIST Each unit includes the following components Injector Head with Cord Control Box Power Supply with Cord 1 vial 3 1 2 Capillaries 1 vial 7 Capillaries 1 Replacement O Ring Kit 1 Allen Wrench 1 2 needle Soe meaner Pp OPERATION The injector head should be mounted in a micromanipulator Brinkman model MM 33 is suitable The aluminum bar rel assembly maximum OD11 mm provides an ideal clamping surface This equipment is ready for use as received it does require pulled capillary tips NVI 570V operation depends greatly on the use of tips prepared from the glass provided We cannot guarantee that glass capillaries with other dimensions will work Pulled capillary tips must be backfilled with oil or other non compressible fluid before attachment on injector Silicone or mineral oil as well as water are commonly used Backfilling is facilitated by using a 30 gauge needle 2 long or disposable spinal needle The NVI 570V will not operate properly without oil The three control switches work as follows FILL Reverses plunger movement while button is depressed Buzzer will sound at extreme fill position Fill speed is 46 nL sec EMPTY Plunger moves fo
4. body the small hole O ring goes first followed by the white plastic spacer with recess facing the tip followed by the O ring with the larger hole The collet screws secure everything C Gasket Replacement The gaskets holding and sealing the capillary tips need periodic replacement OPTIONS AND REPLACMENT PARTS Catalog No Product 65 0036 65 0038 65 0099 65 0037 Foot Switch provides hands off control This Injector is provided with a 3 5 mm jack to allow use of this Foot Switch or other remote control devices Replacment Wire Plunger 3 1 2 Capillaries pkg of 100 2 needle
5. he wire plunger when necessary If the wire plunger is bent it should be replaced for opti mal operation of the unit To change the plunger remove the collet O rings and white spacer The plunger wire can be replaced if accidentally bent This wire is held in place by two set screws Access by removing the black aluminum barrel Hold the motor housing in one hand and unscrew the aluminum barrel with the other hand turn counter clockwise Do not remove the adjacent large black nut Servicing the Unit Contd CAUTION IF YOUR UNIT HAS A HEX NUT BETWEEN THE ALUMINUM BARREL AND THE MIDDLE SECTION DO NOT UNSCREW THIS NUT HOLD THE NUT AND UNSCREW THE ALUMINUM BARREL FROM IT DO NOT REMOVE THE ADJACENT LARGE BLACK NUT IMPORTANT Correct sequence and orientation of the O rings and spacer is critical for proper operation of the injector see next section and figure 1 on previous page Any other service functions should only be attempted after calling our technical service department at 800 232 2380 To Change O Rings After a period of time and or heavy use some leakage of oil during injection might be observed near the collet To correct this the O rings must be replaced A new O ring kit is included with each NVI 570V Refer to figure 1 on previous page since the proper installation is critical to prevent leaking of the unit during injection Proper sequence of the two black O rings and the white spacer is as follows From the pipet
6. rward while button is depressed Buzzer will sound at extreme position Empty speed is approximately twice as rapid as fill i e 110 nL sec A faster speed 230 nL sec can be obtained in fill and empty modes To fill faster hold down the fill button and press the empty button once To empty faster hold down the empty button and press the fill button once The plunger will maintain the faster speed until the control button is released The unit will always start in the slower speed INJECT Depressing button injects a predetermined volume of solution GETTING STARTED A Plunger Home Position Before using the injector the wire plunger must be in the home position When in the home position the tip of the wire plunger will be located even with or slightly recessed from the end of the collet when the collet is tightened When the injector head is plugged into the control box and the power supply is connected the injector will assume it is in the home position If it is not in this position you will not be able to fill the micropipettete to its maximum capacity of 5 ul NOTE UNIT IS SHIPPED FROM THE FACTORY IN THE HOME POSITION GETTING STARTED Contd To reposition the wire plunger simply connect the injector and the power supply to the control box If the wire plunger is extending beyond the collet press and hold the FILL button until the plunger retracts to the end of the collet then release the
7. ss oil out of the micropipette tip Hold the EMPTY button until a double beep is heard The plunger is now fully extended To check measure with a ruler The wire plunger should extend approximately 1 1 16 26 mm from the end of the collet Check for the presence of air bubbles in the micropipette None should be present since they could influence the injection procedure NOTE IF AIR BUBBLES ARE PRESENT REPEAT THE BACKFILLING PROCEDURE C Filling the Micropipette with a Sample After installing the micropipette on the injector place the tip into the sample and hold the FILL button until the audible beep is heard Maximum volume the micropipette will contain is 5 ul If maximum volume is not required release the FILL button and filling will stop D Fast FILL and EMPTY The standard FILL rate is 23 nL sec The standard EMPTY rate is 46 nL sec A faster speed of 230 nL sec can be obtained in both fill and empty modes To fill fast hold down the FILL but ton and press the EMPTY button once To empty fast hold the EMPTY button and press the FILL button once NOTE THE PLUNGER WILL MAINTAIN THE FASTER SPEED UNTIL THE CONTROL BUTTON IS RELEASED THE UNIT WILL ALWAYS START IN THE SLOWER SPEED E INJECT Button Once the micropipettete is filled installed and injected into the oocyte pushing the INJECT button one time will dispense the programmed vol
8. ume By depressing the inject button a second time the same amount of sample will be dispensed again INJECTION RATE 23 nL sec DIP SWITCH Located on side of control box The following code permits the operator to select any of the following 16 volumes VOLUME 1 4 6 nL 9 2 nL 13 8 nL 18 4 nL 23 0 nL 27 6 nL 32 2 nL 36 8 nL 41 4 nL 46 0 nL 50 6 nL 55 2 nL 59 8 nL 64 4 nL 69 0 nL 73 6 nL Dip Switch Cle Oke Cle Oe Che Oke che Oly E cE CEE opu opa CEE CEE opa T Rad E Cle cig cia Che o p ORS Ole OR D U D U D U D U D U D U D U D U GENERAL COMMENTS 1 Wire plungers on all injectors have a point on one end This wire can be reversed if a blunt end is desired The point should allow bubbles in the oil to escape when backfilling tips 2 The preferred tip size seems to be 20 microns Some users report using 30 micron tips with no increase in mortality 3 Injections of 25 nL or less generally requires smaller tips normally 10 microns 4 A broken tip seems preferable for oocyte injection Sharp polished tips tend to skip 5 Virtually any oil is suitable for backfilling 6 The use of a plastic screen in a petri dish is commonly used to hold eggs for injection Also reported is the use of a layer of silicone silastic in a petri dish with a V groove cut after hardening 7 A reported technique positions a drop of mercury between the oil and the sample in the micropipette This
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