V&P SCIENTIFIC LEVITATION STIR BALL LOADER VP 725C-96 Operating Instructions TECHNICAL NOTE 178 9823 Pacific Heights Boulevard, Suite T, San Diego, CA 92121 (858) 455-0643 Fax: (858) 455-0703 email: sales@vp-scientific.com
PREPARATION Warning: Do not make adjustments or place fingers in the machine while it is operating. Remove and sterilize Ball Hopper and Loading Plenums by completing the following steps: 1. Close the Air Valve (1) by turning counter-clockwise (Fig. 1). Bleed air pressure by pushing up on the Pressure Relief Valve (2) (Fig. 1). 2. Unscrew the two Locking Knobs (4) to release the Spring-loaded Bars (14) from the Ball Hopper Base (29) (Figs. 3 & 4). 3. Lift vertically to remove the Ball Hopper (5) and Ball Hopper Base (29) (Figs. 2, 3 & 4). 4. Place in autoclave wrap and autoclave on dry cycle. NOTE: It is very important that the plenums be dry before the balls are added Assemble sterilized Ball Hopper and Loading Plenums onto Ball Loader: 1. Remove autoclave wrap and aseptically place the Ball Hopper (5) (Fig. 2) and Ball Hopper Base (29) (Fig. 4) onto the Loader Platform (9) (Fig. 5). 2. Align the four notches (7) on the Ball Hopper Base to the four dowel pins (8) on the Loader Platform (9) (Figs 9 & 5). NOTE: It is very important that the Yoke (10) of the Ball Rake (11) be aligned to fit in the groove (12) on the Plenum Pneumatic Cylinder (13) (Fig 6). The best way to assure that Yoke (10) is in the right position is to adjust the Yoke position so the end of the Ball Rake (11) is even with the opposite end of the Loader Platform (9) (Figs. 5 & 6). 3. When the Ball Hopper Base (29) is in position, push in Spring-loaded Bars (14) and tighten the Locking Knobs (4) to lock in position (Fig. 6). 4. Connect Ball Loader to air line at Air Line Connector (15) and open Air Valve (1) (Fig. 1). Adjust pressure on the Air Pressure Gauge (41) to 80 to 90 PSI by raising the Cap (16) on Adjustment Knob (17) and then turning the knob (Fig. 1). NOTE: It is very important to operate at this pressure range. 5. Before loading Levitation Stir Balls, place an empty deep well 96 microplate in the Microplate Nest (18) in front of Microplate Pneumatic Cylinder (19) (Figs. 3 & 5). Perform a test by pushing Green Start Button (20) to ensure the pneumatic cylinders are working, the microplate is correctly aligned to the Ball Hopper (5) and the Limit Switches (39, 40) are functioning (Figs. 3 & 5). Repeat the cycle 3-4 times. NOTE: Do not push green button without placing microplate in the nest. If you do, the Microplate Pneumatic Cylinder will stall in the open position. If the microplate is not placed flat in the Microplate Nest (18) it will jam against the Ball Hopper Base (29) when the Green Start Button (20) is pushed (Fig. 3). To reset the Microplate Pneumatic Cylinder (19), manually trigger the two Limit Switch Air Valves (39 & 40) at the rear of the Levitation Ball Loader (Figs. 5 & 7). In order to access the switches remove the Ball Hopper Base (29) from the Loader Platform (9) (Fig. 5) as described above. 6. Make sure that the Mid Plate Plenum (21) (Fig. 8) and the Ball Rake (11) (Fig. 4) are being moved by the Plenum Pneumatic Cylinder (13). V&P Scientific, Inc. 2 Technical Note 178
OPERATION 1. Remove the four Thumb Screws (23) holding the Ball Hopper Lid (26) (Fig. 3) and fill the Ball Hopper (5) with clean, sterile, demagnetized Levitation Balls. Pour the Levitation Balls slowly at first because they bounce very high (Fig. 7). 2. Place a sterile deep well 96 microplate in the Microplate Nest (18) and push the Green Start Button (20) (Fig. 3). 3. Repeat the cycle three times in the same plate to allow the Ball Rake (11) to completely fill the Teflon Loading Plenum (25) (Fig. 8). 4. Remove the first microplate and dump the Levitation Stir Balls back into the Hopper. Re-attach the Ball Hopper Lid (26) (Fig. 7). 5. Replace the first microplate into the Microplate Nest (18) and press the Green Start Button (20) (Figs. 3 & 5). Examine the plate to confirm that every well was loaded with a Stir Ball. 6. Repeat cycle until the level of Levitation Stir Balls in the Ball Hopper goes down to within 1/2 inch of the Teflon Loading Plenum (25) (Fig 3). NOTE: Do not allow the hopper to run below this level. 7. Refill the Ball Hopper with sterile balls (Fig. 7) and keep repeating the cycle. 8. As long as aseptic conditions are maintained in the Ball Hopper, it can be re-filled with sterile Levitation Stir Balls and more plates can be loaded. DISASSEMBLY & MAINTENANCE Disassemble Ball Hopper and Loading Plenums: Caution: Perform this only if absolutely necessary, such as if Loading Plenums are in obvious need of cleaning or if Loader is not functioning properly and might be jammed with a ball or other object. 1. Remove the Thumb Screws (23) on the Ball Hopper Lid to remove the lid (Fig. 3). 2. Unscrew the two Locking Knobs (4) to release the Spring-loaded Bars (14) and the Ball Hopper and Loading Plenums (21 & 25) (Figs. 5 & 6). 3. Lift vertically to remove the Ball Hopper (5) and Ball Hopper Base (29) which will include the Loading Plenums (21 & 25) (Fig. 4). 4. Pour the remaining Levitation Stir Balls into a vessel. 5. Remove the four screws (27) holding the Ball Hopper Walls (28) to the Ball Hopper Base (29). Vertically lift the Flange Frame (30) and the Ball Hopper Walls (Fig. 6) from the Base. (Fig. 8) 6. Lift off the Ball Rake (11) (Fig. 9). 7. Pull the spring-loaded Plenum Retaining Pin (31) and twist it to hold in the open position (Fig. 10). 8. Pull out the Mid Plate Loading Plenum (21) (Fig. 10). V&P Scientific, Inc. 3 Technical Note 178
9. Lift the Teflon Loading Plenum (25) and the Lower Plate Plenum (33) out of the Ball Hopper Base (29) (Fig. 11). Caution: The plenums and base have very close tolerances. For ease, place a microplate under and apply equal pressure to push the plenum out. During disassembly and assembly make certain that the plenums do not become twisted relative to the Ball Hopper Base. If this does occur, gentle tapping with a wooden object is helpful to readjust. 10. Gently pry the Teflon Plenum (25) from the dowel pins holding it to the Lower Plate Plenum (33). Clean Loading Plenums and Ball Rake: 1. Visually inspect all the Loading Plenums and the Ball Rake for damage or wear. 2. Sonicate the Loading Plenums and the Ball Rake in a mild detergent (such as Micro 90 ), and use a cut off Q Tip shaft (not the tip) to clean all the holes. Rinse with tap water, then distilled water and then ethanol. 3. Examine under dissecting microscope to make sure there is no debris blocking the holes. Maintenance checks: 1. Check Break Away Connection (36) from Plate Car (37) to ensure that it will release if the microplate gets stuck under the Loader Base (Fig. 12). 2. Check the integrity of the incoming air line (22) (Fig. 2). 3. Check the Limit Switch Air Valve (39) which is activated by the Microplate Pneumatic Cylinder (19) (Figs. 4, 5 & 7). 4. Check the Limit Switch Air Valve (40) which is activated by the Plenum Pneumatic Cylinder (13) (Figs. 3, 5 & 7). 5. Check to insure the Air Pressure Gauge (41) is functioning by raising Cap (16) and turning Adjustment Knob (17) (Fig. 1). 6. Check that the Time-Delay Air Valve for Plenum Pneumatic Cylinder (42) has a 6 second delay, (Fig. 13). 7. Check that the Time-Delay Air Valve for Microplate Pneumatic Cylinder (43) has a 1-second delay (Fig. 13). 8. Check the integrity of the air lines entering and leaving the Directional Air Valve (44) and the Air Line Manifold (45) (Fig. 13). REASSEMBLY 1. Reassemble the Ball Hopper and Loading Plenums by reversing disassembly steps 1 through 10 in previous section. 2. When mating the Teflon Loading Plenum (25) to the Lower Plate Plenum (33) (Fig. 11), ensure that the Ball Rake Groves in the Teflon Loading Plenum are facing up. Also make sure that the lip end (bottom side of Teflon Loading Plenum) is at the same end of the Lower Plate Plenum containing the four notches (Fig. 11). 3. Place the 2 Plenums inside the Ball Hopper Base (29) (Fig. 11). Caution: The plenums and base have very close tolerances. During disassembly and assembly certain that the plenums do V&P Scientific, Inc. 4 Technical Note 178
not become twisted relative to the Ball Hopper Base. If this does occur, gentle tapping with a wooden object is helpful to readjust. 4. The moveable Mid Plate Plenum is then inserted on the side with the Plenum Retaining Pin (31) (Fig. 10). When inserting the moveable Mid Plate Plenum (21) between the Teflon Loading Plenum (25) and the Lower Plate Plenum be sure that the lettering TOP remains on the upper (Teflon) side. 5. Turn the Plenum Retaining Pin (31) until it clicks and locks the moveable Mid Plate Plenum (21) inside (Fig. 10). 6. Place the Ball Rake (11) in the grooves on the Teflon Loading Plenum (25) (Fig. 9). 7. Align the four notches (7) on the Ball Hopper Base to the four dowel pins (8) on the Loader Platform (9) (Figs 9 & 5). NOTE: It is very important that the Yoke (10) of the Ball Rake (11) be aligned to fit in the groove (12) on the Plenum Pneumatic Cylinder (13) (Fig 6). If the Yoke is adjusted so the end of the Ball Rake is even with the opposite end of the Ball Hopper Base, the Yoke will fit into the groove (Fig. 6). 8. Place the Ball Hopper Walls (28) on to the Ball Hopper Base (29) (Fig 4). 9. Place the Flange Frame (30) over the Ball Hopper Walls (28) and fasten with the four screws (27) (Fig. 3). 10. Press in the Spring-loaded Bars (14) and tighten the two Locking Knobs (4) to attach the Ball Hopper and Loading Plenums to the Loader Platform (Fig 3 & 4). 11. The Levitation Ball Loader is now ready to operate, connect to compressed air line and adjust pressure to 80 to 90 PSI. Continue following instructions in Operations section. FIGURES: 3-- V&P Scientific, Inc. 5 Technical Note 178
29 V&P Scientific, Inc. 6 Technical Note 178
9 9 11--- V&P Scientific, Inc. 7 Technical Note 178
11 29 29 V&P Scientific, Inc. 8 Technical Note 178
V&P Scientific, Inc. 9 Technical Note 178 29
PARTS LEGEND [Found in Figure #] (1) Air Valve [1] (2) Pressure Relief Valve [1] (3) On/Off Switch [1] (4) Locking Knob [3,5,6] (5) Ball Hopper [2,7] (6) Cover [2] (7) Ball Hopper Alignment Notches [4,9] (8) Dowel Pins on Loader Platform (9) Loader Platform [5] (10) Ball Rake Yoke [6, 8, 13] (11) Ball Rake [4, 8, 9] (12) Groove on the Plenum Pneumatic Cylinder [5, 6] (13) Plenum Pneumatic Cylinder [4, 5, 6, 13] (14) Spring-Loaded Bars [3,5,6] (15) Air Line Connector [1] (16) Air Pressure Adjustment Knob Cap [1] (17) Air Pressure Adjustment Knob [1] (18) Microplate Nest [2, 3, 12] (19) Microplate Pneumatic Cylinder [2, 3, 12] (20) Green Start Button [2, 3, 12] (21) Mid Plate Loading Plenum [8,9] (22) Airline [2] (23) Ball Hopper Lid Thumb Screws [3] (24) Bottle of Sterile Stir Balls [7] (25) Teflon Loading Plenum [8, 9, 10, 11] (26) Ball Hopper Lid [4, 6, 8] (27) Ball Hopper Flange Frame Screws [6, 7] (28) Ball Hopper Walls [4, 6, 8] (29) Ball Hopper Base [3, 4, 6, 8, 9, 10, 11] (30) Ball Hopper Flange Frame [3, 6, 8] (31) Spring Loaded Plenum Retaining Pin [10, 13] (32) (33) Lower Plate Loading Plenum [8, 11] (34) Plate Car Guide Rails [5] (35) (36) Break Away Connection [12] (37) Plate Car [3] (38) Microplate Bumper Stops [5] (39) Limit Switch Air Valve, Activated by Microplate Pneumatic Cylinder [4, 5, 7] (40) Limit Switch Air Valve, Activated by Plenum Pneumatic Cylinder [3, 7] (41) Air Pressure Gauge [1] (42) Time-Delay Air Valve for Plenum Pneumatic Cylinder[13] (43) Time-Delay Air Valve for Microplate Pneumatic Cylinder [13] (44) Directional Air Valve [13] (45) Air Line Manifold [13] V&P Scientific, Inc. 10 Technical Note 178