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ProCulture® Spinner Flasks

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1. Bend the paddle blades slightly and insert the assembled paddle into the center neck of the flask Fig 18 After insertion straighten the paddle blades with your fingers Also re insert the tabs on the top of the paddle blade into the notches on the paddle blade retaining sleeve Fig 19 Then screw down the cap on the threads of the flask center neck Fig 20 Step 2 Figures 21 and 22 Note the location of the two engagement slots for the bayonet fittings see Fig 17 Dowel Pins 1 8 x 1 on the top of the drive shaft Fig 21 The dowel pins on the bottom of the Direct Drive Motor Fig 22 will slide into these slots to secure the motor in place for operation Step 3 Figure 23 Tilt the motor slightly from vertical and slide the yoke coupling into the cavity on the top of the direct drive shaft Fig 23 The yoke dowel pins will engage with the drive pin inside of the cavity when the motor is correctly oriented Figure 18 Insert the assembled paddle into Figure 19 Straighten paddle blades after flask bend paddle if necessary insertion Figure 20 Place cap on flask and tighten PROCULTURE SPINNER FLASKS OPERATING INSTRUCTIONS Cap Assemblies Direct Drive ProCulture Spinner Flasks 4510 and 4512 Series Flasks continued Step 4 Figure 24 Turn the motor until the dowel pins on the motor drive adapter assembly line up with the vertical section of the bayonet fitting You can use the end of the dowel pi
2. Dual angled sidearm fittings can be used for aseptically transferring medium into or out of angled sidearm spinner flasks or for sparging the medium with gases Fittings are comprised of a PET insert with a Viton O ring and a polypropylene sealing cap Two 316 stainless steel tubes which extend to the bottom of the flask are held in place by Noryl nuts with integrated ferrules The fittings are completely autoclavable Gas or Media Handling Fittings Angled Sidearm Flasks Dual Style Ordering Information Flask Tubing O D Qty Cat No Description Size inches Case 4519 150 SA fitting Dual IL 1 8 1 4519 151 SA fitting Dual 3L 1 8 I 4519 152 SA fitting Dual 6L 1 8 I 4519 153 SA fitting Dual 8L 1 8 I 4519 173 SA fitting Dual IL 1 8 1 4 I 4519 121 SA fitting Dual 8L 1 8 1 4 1 4519 174 Sidearm fitting Dual 500mL s angled to 125 mL level 4 1 4519 154 Sidearm fitting Dual IL 1 4 1 4519 155 Sidearm fitting Dual 3L 1 4 1 4519 156 Sidearm fitting Dual 6L 1 4 l 4519 157 Sidearm fitting Dual 8L 1 4 1 4519 170 Sidearm fitting Dual 15L 1 4 1 Gas or Media Handling Fittings Angled Sidearm Flasks Combination Style Used to aseptically transfer medium sparge the cell culture medium directly or add gases to the head space above the cell culture medium Fittings are comprised of a PET insert with a Viton O ring and a polypropylene sealing cap One or two 316 stainless steel tubes extend t
3. Step Compression Sealing Gasket Step 1 Y Center Shaft Lower Plate Step 7 Shaft Retaining Nut with Integrated m Ferrule Completed Paddle Assembly and Retaining Shaft Figure 11 Magnetically Driven Shur Loc Cap Assembly Cap Assemblies Magnetically Driven ProCulture Spinner Flasks Continued Shur Loc Style Caps for 4504 Series Flasks and for Production Style 4500 and 4502 Series Flasks Figure 11 The Shur Loc style cap is composed of Lower Plate Upper Plate Compression Sealing Gasket Seal Actuating Ring Washer Retaining O Ring Shaft Retaining Nut Step 1 D D D Seal Actuating Ring D D D Slide the large Compression Sealing Gasket onto the Lower Plate until it fits over the notched area Step 2 Insert the threaded Center Shaft of the Lower Plate into the hole in the Upper Plate Step 3 Hold the two halves together and screw the Seal Actuating Ring loosely onto the extending threaded Center Shaft Step 4 Drop the Seal Actuating Ring Washer onto the shaft and then push the Retaining O ring onto the shaft until the O ring fits into the notch on the shaft Step 5 Turn the Seal Actuating Ring clockwise until you feel the large Compression Sealing Gasket begin to extend out from the cap assembly Step 6 Slide the Shaft Retaining Nut with the knurled larger end first onto the glass polysulfone or stainless steel shaft of the Completed Pad
4. 402614 400649 100 120 mm Cap Assembly Direct Drive Style Complete for ProCulture Direct Drive 100 120 mm Cap 100 120 mm Cap Liner Direct Drive Shaft Assembly S S Coupling Collar I5 XXX Direct Drive Paddle Assembly Complete with S S Shaft Direct Drive Paddle Bearing Holder Direct Drive Paddle Blade Direct Drive Paddle S S Rod Direct Drive Paddle Retaining Sleeve Direct Drive Paddle Retaining Plug eG Pe Oro Corning USA Figure 36 ProCulture 4510 12 Series 100 120 mm Center Neck Flask Fitted with 402614 400649 Direct Drive PROCULTURE SPINNER FLASKS OPERATING INSTRUCTIONS Spinner Flask Components Chemical Compatibility Chart Chemical Resistance FDA Acid Base Component Composition Compliant Autoclavable 1 N HCl 1 N NaOH Organics Flask Glass Corning 7740 Yes Yes Yes Yes Yes Type I Borosilicate Cap Assemblies Shur Loc Head Plates Polypropylene Yes Yes Yes Yes Center Neck Screw Caps Valox 420 Yes Yes Yes Yes Sidearm Screw Caps Polypropylene Yes Yes Yes Yes Shaft Retaining Nut ULTEM 1000 Yes Yes Yes Yes Shur Loc Head Plate Compression Sealing Gasket Silicone Yes Yes Yes Yes O ring Silicone Yes Yes Yes Yes Seal Actuating Ring PET Yes Yes Yes Yes Seal Actuatin
5. 1 Ashing Glassware can be treated by baking at a temperature of at least 450 C 840 F for at least 3 hours This removes all organic compounds to a mineral and carbon ash DO NOT USE THIS TREATMENT ON ANY OF THE PLASTIC OR SILICONE SYNTHETIC RUBBER COMPONENTS 2 Chemical treatment with NaOH Follow the procedure described above under Depyrogenation 3 Chemical treatment with Chromic Acid This treatment is very effective in decomposing biological molecules and also is helpful to reduce mineral deposits Be sure to follow all normal safety precautions when handling acid solutions Chromic acid is made by dissolving 20 g of Na or K chromate in sufficient water to make a paste Add 300 mL of concentrated H SO Glassware can be treated for as long as 24 hours Rinse and clean afterwards as described in the Standard Glassware Cleaning Protocol on page 7 Removal of Mineral Deposits Residual scale which is built up on glassware may be removed to varying extents by treatments with acids such as sulfuric nitric hydrochloric acid or chromic acids as described above The nitric acid treatment is especially useful in removing buildup of material causing corrosion of stainless steel parts Follow safety procedures in handling as with chromic acid above Start with short mild treatments and gradually increase the strength of the reagent and length of exposure as necessary PROCULTURE SPINNER FLASKS OPERATING INSTRUCTIONS Addi
6. Magnetically Driven Paddle Assemblies Step 4 Figure 4c Slide the Paddle Blade into the notch in the Bearing Holder narrow end of the paddle blade first until the Hole for the Magnet in the Paddle Blade lines up with the Hole for the Magnet in the Bearing Holder Step 5 Figure 4c and d Take the Blade Retaining Sleeve with the notched end facing down and slide it down over the top of the Glass PolySulfone or Stainless Steel Shaft and onto the Bearing Holder Paddle Blade Assembly Position the narrow ends of the Paddle Blade into the notches of the Blade Retaining Sleeve and push down on it until the top portions of the Paddle Blade extend completely into the notch of the Blade Retaining Sleeve Step 6 Figure 4d Insert the Magnet into the aligned holes until it is centered in the com bined assembly The Paddle Blade should not be able to slide on the Bearing Holder It should now be held in place by the Magnet Note If the narrow portion of the Paddle Blade should disengage from the Blade Retaining Sleeve during subsequent handling of the paddle assembly simply place it back into the notch by gently twisting to insert it into the notches on the retaining sleeve PROCULTURE SPINNER FLASKS OPERATING INSTRUCTIONS Step 5 gt Stainless Tighten Steel Set Screws Coupling gt i Collar Step 4 i Stainless Steel Shaft Blade Retaining SI Step 3 eeve Bearing Holder Step1 Paddle Blade Step 2 Retaining P
7. Paddle Blade Assembly Figure 4 Magnetically Driven Paddle Assemblies PROCULTURE SPINNER FLASKS OPERATING INSTRUCTIONS Flasks with Glass Shafts Figure 4 a c d Step 1 Figure 4a Remove the PTFE Split Ring from the end of the glass Shaft Save this split ring for later use It will be used in the final assembly of the cap Step 2 Figure 4a Slide the end of the shaft that is missing the PTFE split ring into the hollow Bearing Holder Step 3 Optional Figure 4a Insert the Silicone Plug into the Bearing Holder This will safe guard against mircocarriers getting up into the Bearing Holder Proceed to Steps 4 6 Flasks with PolySulfone Shafts or Optional Stainless Steel Shafts Figure 4 b c d Step 1 Figure 4b Remove one PTFE Split Ring from either end of the PolySulfone or Stainless Steel Shaft These shafts have a hole drilled down through the center of the shaft and a small hole in the side of the shaft The holes are interconnected and ensure that steam enters the Integrated Compression Fitting in the cap during autoclaving Save this split ring for later use It will be used in the final assembly of the cap Step 2 Figure 4b Slide the end of the shaft that is now without the PTFE Split Ring into the hollow Bearing Holder Step 3 Optional Figure 4b Insert the Silicone Plug into the Bearing Holder This will safe guard against mircocarriers getting up into the Bearing Holder Proceed to Steps 4 6 All
8. angled sidearm flasks the probes and certain operations can be mounted on the center neck cap and further operations located on the sidearms On vertical sidearm flasks the sensors and most of the accessories are situated on the sidearms and the center neck cap can be modified with smaller tubing for venting or additions of supplements By combining sidearm fittings with the angled or vertical sidearm flasks and system con trollers sophisticated bioreactor operation can be easily achieved at a price considerably below the cost of buying an off the shelf unit from a fer menter manufacturer The Unit Operations that can be created include supplying or controlling additional gases i e oxygen to flasks in order to improve the exchange of the gas with the liquid medium adding and or removing fresh medium in fed batch operations to replen ish spent nutrients in the medium performing in line filtration of the culture medium using microporous filters to remove spent medium which may inhibit cell growth adding sensors and liquid addition ports into the vessel to help monitor and or control pH O or temperature inoculating the cell culture medium at the beginning or during a cell culture process For the larger vertical sidearm flasks Corning offers overhead direct drive motors and drive shafts see next section for more consistent agitation with the larger volumes in the flasks To determine the configuration of the flask and accesso
9. end user Why choose an augmented and or bioreactor process Augmenting your spinner flask for headspace overlays direct sparging media transfer and or pH and oxygen monitoring is mainly considered in order to increase cell density and productivity in the larger flasks As cell culture volume increases a decrease in cell density may occur due to factors such as nutrients and oxygen becoming limited in the flasks While Corning spinner flasks are better than conventional flasks in dealing with these issues additional operations are still needed to help overcome these limitations and boost productivity What type of augmented operation should I use There are a number of easy solutions to some of these limitations A Corning Specialist can help you in configuring your flasks for these operations The options available to you are detailed at the back of this manual in the Ordering section for sidearm fittings starting on page 35 PROCULTURE SPINNER FLASKS OPERATING INSTRUCTIONS 26 Optional Operation Sidearm Fitting Accessories and Center Neck Cap Modifications Corning provides a unique opportunity for achieving better performance from your spinner flasks Accessories are available for both angled sidearm flasks and vertical sidearm flasks for augmented operations such as head space overlays direct sparging media transfer and pH and oxygen moni toring Positioning of the accessories is dependent on the desired unit operation On
10. mm Center Neck Opening 15000 mL Replacement Flask w 140 mm Center Neck Opening 36000 mL Replacement Flask Spinner 100 mm Center Neck 8000 mL Four 45 mm Vertical Sidearms Replacement Flask Spinner 100 mm Center Neck 15000 mL Four 45 mm Vertical Sidearms Replacement Flask Spinner 100 mm Center Neck 36000 mL Six 45 mm Vertical Sidearms Replacement Flask Spinner 120 mm Center Neck 8000 mL Four 45 mm Vertical Sidearms Replacement Flask Spinner 120 mm Center Neck 15000 mL Four 45 mm Vertical Sidearms Replacement Flask Spinner 120 mm Center Neck 15000 mL Six 45 mm Vertical Sidearms Replacement Flask Spinner 120 mm Center Neck 36000 mL Six 45 mm Vertical Sidearms BP 3700 314 BP 3700 316 Replacement Flask Spinner 140 mm Center Neck 15000 mL Four 45 mm Vertical Sidearms Replacement Flask Spinner 140 mm Center Neck 36000 mL Six 45 mm Vertical Sidearms Impeller Assemblies Complete BP 3401 100 BP 3401 102 BP 3401 104 BP 3401 106 BP 3401 108 BP 3401 110 BP 3401 112 BP 3401 114 BP 3401 116 Impeller w PS Shaft Magnetic Complete 125 mL Impeller w PS Shaft Magnetic Complete 250 mL Impeller w PS Shaft Magnetic Complete 500 mL Impeller w PS Shaft Magnetic Complete 1000 mL Impeller w Glass Shaft Magnetic Complete 3000 mL Impeller w Glass Shaft Magnetic Complete 6000 mL Impeller w Glass Shaft Magnetic Complete 8000 mL Impeller w Glass Shaft Magnetic Complete 15000 mL Impeller w Gla
11. more readily through the sidearms Autoclaving l After assembly place the vessel into an autoclave We recommend that the vessels are autoclaved in an upright position Fig 27 Note A small amount of deionized water may be added to the vessel to generate additional steam during autoclaving Fig 28 Check once more that the sidearm caps are still loosely attached and that the cap on the center neck has not shifted during transport to the autoclave to allow for adequate transfer of steam into the vessel and venting during and after autoclaving If the vessel is too large to autoclave in an upright position the cap and paddle assembly must be removed autoclaved separately in an autoclave bag and then aseptically reassembled in sterile conditions under a hood Failure to do so may result in the bearing holder warping which will result in poor agitation of the paddle assembly Autoclave the vessel using a liquid cycle for a minimum of 30 minutes at 121 C and 15 psi For larger vessels extended autoclaving may be necessary All parts of the vessel have been designed to be autoclaved repeatedly using the conditions described above Allow the vessel to cool after autoclaving On vessels with the Shur Loc center neck opening Corning flasks 4504 series and 4514 series with 140 mm center neck openings or Production Style 4500 series and 4502 series it is advisable to turn the seal actuating ring on the c
12. securing 45 mm 22 PTFE grey Fitting nut Fitting O ring Fitting insert a TT Fitting O ring Spinner Flask Shaft Retrofit Kits Retrofit Kits Dual Bearing Shafts Polysulfone or Glass BP 3401 400 BP 3401 401 BP 3401 402 BP 3401 403 BP 3401 404 BP 3401 405 BP 3401 406 BP 3401 407 BP 3401 408 Retrofit Kit Polysulfone Shaft 125 mL Retrofit Kit Polysulfone Shaft 250 mL Retrofit Kit Polysulfone Shaft 500 mL Retrofit Kit Polysulfone Shaft 1000 mL Retrofit Kit Glass Shaft 3000 mL Retrofit Kit Glass Shaft 6000 mL Retrofit Kit Glass Shaft 8000 mL Retrofit Kit Glass Shaft 15000 mL Retrofit Kit Glass Shaft 36000 mL Retrofit Kits Dual Bearing Shafts Stainless Steel BP 3601 400 BP 3601 401 BP 3601 402 BP 3601 403 BP 3601 404 BP 3601 405 BP 3601 406 BP 3601 407 BP 3601 408 Retrofit Kit Stainless Steel Shaft 125 mL Retrofit Kit Stainless Steel Shaft 250 mL Retrofit Kit Stainless Steel Shaft 500 mL Retrofit Kit Stainless Steel Shaft 1000 mL Retrofit Kit Stainless Steel Shaft 3000 mL Retrofit Kit Stainless Steel Shaft 6000 mL Retrofit Kit Stainless Steel Shaft 8000 mL Retrofit Kit Stainless Steel Shaft 15000 mL Retrofit Kit Stainless Steel Shaft 36000 mL For ordering and technical service of all BP 3xxx xxx part numbers please call Wilmad LabGlass at 1 800 220 5171 ext 686 in the U S Outside the U S please call 001 856 697 3000 ext 686 For all other part numbe
13. than 8L sizes especially at the 15L and 36L sizes Magnets in stirrer plates are too small and thus do not have enough power to stir large paddles and the increased volumes of liquid that are in the larger flasks The direct drives couple the drive shaft and motor together to aid in better stirring Which manufacturer probes can I use and which kind For autoclavable insertion style oxygen pH or temperature sensor probes Broadley James and Ingold are the two major manufacturers of these probes For sizes ranging from 500 mL to 8L flasks most investiga tors use 12 mm diameter probes with a length long enough to get the tip of the probe to approximately 50 of the working volume of the flask When using the 15L and 36L size flasks older 25 mm diameter probes can also be used Why choose vertical sidearms rather than angled sidearms The choice of whether to use an angled or vertical sidearm is mainly based on versatility and ease of operation It is easier to operate flasks and to position glass and stainless steel sensor probes into the medium of flasks with vertical sidearms than those with angled sidearms Introducing gases into a flask with a tube positioned in a vertical orientation is easier than bending a tube through an angled sidearm configuration In addition when side arms are in a vertical orientation more sidearms can be added and thus more fittings can be utilized in a vertical orientation creating more options for the
14. the U S please call your local Corning Representative to determine the availability of side port fittings to match the type of probe and fitting If a probe is not an insertion style there is a nominal fee for specially modifying side ports to match your needs Contact your local Corning Representative PROCULTURE SPINNER FLASKS OPERATING INSTRUCTIONS 7 Select liquid handling and gassing requirements if any Liquid handling and gassing tubes can be obtained in 1 8 1 4 or 1 2 OD sizes These sizes are roughly matched to fit the 8L 15L and 36L vessels respectively but should be used only as a guide For some applica tions an inlet tube of greater or lesser diameter may be more appro priate In these cases it is often possible to mix and match tubing of differing diameters on the same fitting The 4519 120 122 fittings are an example of such a situation In most cases 1 2 OD tubing will be necessary for any microporous filtration applications to provide sufficient recirculation flow to gen erate the flux rates necessary for effective tangential flow filtration Flask venting can be accomplished with two types of fittings 1395 45LTMC Features an integral 25 mm 0 2 micron PTFE fil ter built into the cap This is generally suitable for operation with 1 8 inlet gassing fitting 4519 106 Features a threaded design which will accept most replaceable 50 mm 0 2 micron PTFE filters with 1 8 NPT thr
15. Assembly To do so grasp the housing firmly and turn the bottom plate counter clockwise until it turns Step 2 Take the housing and top plate portion of the Bellco drive assembly and gently screw into the threaded opening of the 100 or 120mm cap Support the silicone liner on the underside of the cap to keep it from being displaced This will require a number of turns but ultimately the shaft housing will bottom out in the cap assembly and stop turning The drive shaft should protrude out of the bottom of the cap assembly Step 3 Slide the assembled coupling collar paddle assembly up onto the protruding drive shaft of housing Step 4 Tighten the set screws on both sides of the collar so that the collar does not turn on the drive shaft Consult your Bellco Instruction Manual for the OverHead Drive if you have any additional questions regarding the Bellco Drive PROCULTURE SPINNER FLASKS OPERATING INSTRUCTIONS Figure 27 Place spinner flask in an upright position when autoclaving the vessel Do not lay on its side Figure 28 A small amount of water 5 10 mL of distilled or deionized water per 10 L of vessel volume can be added into the larger vessel to generate additional steam in the vessel during autoclaving Final Assembly of Vessels and Autoclaving Vessel Assembly Is Insert the combined paddle and cap assembly into the vessel It may be necessary on the larger vessels to bend the paddl
16. ProCulture Spinner Flasks Operating Instructions 4500 Series 4510 Series 4502 Series 4512 Series 4504 Series 4514 Series Life Sciences Webbe Toa Corning Incorporated Life Sciences 45 Nagog Park Acton MA 01720 USA t 1 800 492 1110 1 978 635 2200 f 1 987 635 2476 www corning com lifesciences Copyright 2005 CORNING INCORPORATED All rights reserved No part of this document may be reproduced in any form without the written permission of CORNING INCORPORATED CORNING has taken every reasonable precaution to ensure the accuracy of the information descriptions and instructions in this manual However Corning assumes no responsibility for errors omissions or possible misinterpretations in addition to conse quences resulting from the operation of the equipment based on these instructions CORNING Limited Warranty CORNING INCORPORATED warrants this product against defects in material or workmanship for a period of one year after date of delivery to the Purchaser CORNING DISCLAIMS ALL OTHER WARRANTIES WHETHER EXPRESSED OR IMPLIED INCLUDING WITHOUT LIMITATIONS THE WARRANTY OF MERCHANTABILITY OR OF FITNESS FOR A PARTICULAR PURPOSE Corning s sole obligation shall be to repair or replace at its option any product or part thereof that proves defective in material or workmanship within the warranty period provided the Purchaser notifies CORNING of any such defect Corning shall in no event be liable for consequentia
17. ap slightly clockwise to expand the O ring slightly as the vessel cools since the cap assembly contracts slightly while it cools This will guarantee that the vessel remains sealed and sterile PROCULTURE SPINNER FLASKS OPERATING INSTRUCTIONS Basic Operation Important Note We strongly recommend that a sterility check be accomplished by incubating the vessel containing the medium for 24 hours prior to inoculation of the vessel This will insure that the sterilization and medium transfer were accomplished aseptically Sterile Medium Figure 29 Small vessel being filled through a sidearm Filling the Vessel with Medium This section provides basic information for cell culture methods using ProCulture Spinner Flasks Novices should consult a detailed guide to cell culture methods such as Culture of Animal Cells A Manual of Basic Technique by R Ian Freshney Alan R Liss Publishing New York 1994 or Cell and Tissue Culture Laboratory Procedures by Doyle A et al John Wiley amp Sons Sussex England 1995 for specific details There are also several fine video tapes which describe the methods in detail The websites of media companies are also an excellent resource They contain the most current media under development for cell suspension cultures Filling Filling the vessel can be accomplished in a variety of ways and is best done in a sterile biological hood or a clean room Small vessels Center Nec
18. arger opening on coupling Figure 9 Tighten lower set screw Step 5 collar over paddle shaft Step 4 PROCULTURE SPINNER FLASKS OPERATING INSTRUCTIONS a Shaft Retaining Nut o Step 2 Impeller Shaft Assembled Paddle b Insert O gt Step 3 the PTFE Split Ring Impeller Shaft Screw Cap with Integrated c Compression Fitting T Completed Paddle Assembly and Retaining Shaft Cap Assemblies Magnetically Driven ProCulture Spinner Flasks Screw Caps for 4500 and 4502 Series Flasks Figure 10 a c The 70 mm 100 mm and 120 mm screw cap assemblies are comprised of 3 components Screw Cap with Integrated Compression Fitting ICF Figure 10c Silicone Liner Figure 10c Shaft Retaining Nut Figure 10a Note Glass polysulfone and stainless steel shafts for 4500 4502 and 4504 impeller assemblies come in 2 sizes 6 mm 1 4 for 125 mL 1000 mL flasks and 8 mm 5 16 for all flasks 3000 mL and larger The size of the Shaft Retaining Nut must be identical to the size of the corresponding Impeller Shaft Figure 10a A 6 mm 1 4 Impeller Shaft will only work with a 6 mm 1 4 Shaft Retaining Nut For example it will not be possible to insert a large 8 mm 5 16 shaft into a smaller 6 mm 1 4 hole in the nut In the other instance if a smaller 6 mm 1 4 shaft is inserted into a large 8 mm 5 16 retaining nut there is excessive side to side wobble of the shaft a
19. aving Check the wire brushes used to clean the vessels and discard them when the bristles are worn to avoid scratching the glassware Over time buildup of biological or mineral residues on the walls of the glass vessels or parts may occur If this happens consult the section below entitled Rigorous Glassware Cleaning Protocols for Severely Soiled Flasks The synthetic rubber Delrin and Noryl components will slowly age from repeated autoclaving and become brittle These parts should be replaced when the decrease in their flexibility or stress fractures becomes apparent Plastics especially PTFE have a tendency to warp during autoclaving This is normal but may cause problems after extended periods in the ability of the paddle to spin freely on the glass or stainless steel shafts If this happens a replacement PTFE bearing holder can be purchased to replace the warped one Avoid the use of organic solvents on any of the plastic or synthetic rub ber parts since it may weaken them and significantly reduce their life span Rigorous Glassware Cleaning Protocols for Severely Soiled Flasks After extended use residues may build up on the flasks and components requiring more rigorous cleaning of the flasks These residues may include pyrogenic materials buildup of dead cells and debris on the vessel and components of the cell culture medium To minimize the buildup of these materials we recommend that the parts are soaked in a dilut
20. bers please call Wilmad LabGlass at 1 800 220 5171 ext 686 in the U S Outside the U S please call 001 856 697 3000 ext 686 For all other part numbers please call Corning Incorporated at 1 800 492 1110 in the U S Outside the U S please call your local Corning Representative 40 PROCULTURE SPINNER FLASKS OPERATING INSTRUCTIONS Replacement Parts for Spinner Flasks Paddle Retaining Sleeves BP 3411 050 BP 3411 053 BP 3411 055 BP 3411 057 Paddle Retaining Sleeves 125 mL 500 mL Paddle Retaining Sleeves 1000 mL Paddle Retaining Sleeves 3000 mL 8000 mL Paddle Retaining Sleeves 15000 mL 36000 mL Paddle Magnets BP 3411 060 BP 3411 061 BP 3411 062 BP 3411 063 BP 3411 068 Paddle Magnets for 250 mL amp 500 mL Flask 3 8 x 2 Paddle Magnets for 1000 mL Flask 1 2 x 3 Paddle Magnets for 3000 mL 8000 mL Flask 5 8 x 4 Paddle Magnets for 15000 mL 36000 mL Flask 3 4 x 3 1 4 Paddle Magnets for 125 mL Flask 3 8 x 1 1 2 Paddle Rods Polysulfone or Glass Dual Bearing BP 3411 170 BP 3411 170 BP 3411 172 BP 3411 173 BP 3411 174 BP 3411 175 BP 3411 176 BP 3411 177 BP 3411 176 Paddle Polysulfone Rods 125 mL 6 mm O D Paddle Polysulfone Rods 250 mL 6 mm O D Paddle Polysulfone Rods 500 mL 6 mm O D Paddle Polysulfone Rods 1000 mL 6 mm O D Paddle Glass Rods 3000 mL 8 mm O D Paddle Glass Rods 6000 mL 8 mm O D Paddle Glass Rods 8000 mL 8 mm O D Paddle Glass Rods 15000 mL 8 m
21. bled Coupling Collar Paddle Assembly up onto the protruding drive shaft Step 4 Tighten the Set Screws on both sides of the Coupling Collar so that the collar does not turn on the drive shaft Alternatively slide the smaller opening on the coupling collar over the drive shaft in the cap assembly Fig 13 and tighten the upper set screw Fig 14 Insert the stainless steelpaddle shaft into the larger opening in the bottom of the coupling collar Fig 15 and then tighten the lowerset screw on the coupling collar Fig 16 Figure 13 Insert drive shaft in cap assembly into the smaller opening of the coupling collar Figure 15 Insert paddle shaft into coupling collar Figure 16 Tighten lower set screw on the coupling collar PROCULTURE SPINNER FLASKS OPERATING INSTRUCTIONS A Q ARROW 350 Direct Drive Motor High Torque Clockwise rotation Motor shown for assembly purposes only LIL lose Drive Coupling Yoke Type PB 3921 XXX Coupler Motor Drive Adapter Assembly 18 8 SS BP 3921 605 Yoke Dowel Pins BP 3921 xxx Drive Coupling Yoke Type Dowel Pins 1 8 x 1 Figure 17 Schematic of Direct Drive Motor 14 Cap Assemblies Direct Drive ProCulture Spinner Flasks 4510 and 4512 Series Flasks continued Inserting Assembled Paddle into Flask and Coupling of Motor to Direct Drive Shaft Fig 17 Step 1 Figures 18 19 20
22. cement Parts for Spinner Flasks Replacement Flasks Complete BP 3405 100 BP 3405 102 BP 3405 104 BP 3405 106 BP 3405 108 BP 3405 110 BP 3405 112 BP 3405 114 BP 3405 116 BP 3505 108 BP 3505 110 BP 3505 112 BP 3505 114 BP 3505 116 BP 3605 108 BP 3605 110 BP 3605 112 BP 3605 114 BP 3605 116 BP 3700 112 BP 3700 114 BP 3700 116 BP 3700 212 BP 3700 214 BP 3700 215 BP 3700 216 Replacement Flask w 70 mm Center Neck Opening 125 mL Replacement Flask w 70 mm Center Neck Opening 250 mL Replacement Flask w 100 mm Center Neck Opening 500 mL Replacement Flask w 100 mm Center Neck Opening 1000 mL Replacement Flask w 100 mm Center Neck Opening 3000 mL Replacement Flask w 100 mm Center Neck Opening 6000 mL Replacement Flask w 100 mm Center Neck Opening 8000 mL Replacement Flask w 100 mm Center Neck Opening 15000 mL Replacement Flask w 100 mm Center Neck Opening 36000 mL Replacement Flask w 120 mm Center Neck Opening 3000 mL Replacement Flask w 120 mm Center Neck Opening 6000 mL Replacement Flask w 120 mm Center Neck Opening 8000 mL Replacement Flask w 120 mm Center Neck Opening 15000 mL Replacement Flask w 120 mm Center Neck Opening 36000 mL Replacement Flask w 140 mm Center Neck Opening 3000 mL Replacement Flask w 140 mm Center Neck Opening 6000 mL Replacement Flask w 140 mm Center Neck Opening 8000 mL Replacement Flask w 140
23. d in place by Noryl nuts with integrated fer rules The fittings are completely autoclavable Fittings for Insertion Probes Vertical Sidearm Flasks Ordering Information Sensor O D Qty Cat No Description mm Case 4519 108 Sidearm fitting sensor O probes 12 I 4519 128 Sidearm fitting sensor temperature probes 12 I 4519 110 Sidearm fitting sensor pH probes 12 l 4519 172 Sidearm fitting sensor pH or O 18 1 Impeller Assembly for Magnetically Driven Bioreactor Stainless steel impeller shaft with modified impeller blade for use with probes to create a small bioreactor Impeller Assembly for Magnetically Driven Bioreactor Ordering Information Cat No Description Qty Cs 402648 Impeller assembly stainless steel dual bearing 1 modified for probes 3L 402649 Impeller assembly stainless steel dual bearing 1 modified for probes 6L 401392 Impeller assembly stainless steel dual bearing 1 modified for probes 8L 401661 Impeller assembly stainless steel dual bearing 1 modified for probes 15L 402650 Impeller assembly stainless steel dual bearing 1 modified for probes 36L PROCULTURE SPINNER FLASKS OPERATING INSTRUCTIONS Direct Drive Motor Direct Drive Paddle Assembly Cap Assembly for Magnetically Driven Bioreactor Cap assembly for small biorecator with various fitting arrangements Cap Assembly for Magnetically Driven Bioreactor Ordering Information Cat No Descriptio
24. dle Assembly and Retaining Shaft and replace the PTFE Split Ring back onto the notch on the shaft from which it was previously removed during the Paddle Assembly process page 8 Step 7 Insert the Completed Paddle Assembly and Retaining Shaft into the newly completed Shurloc Cap Assembly and tighten the assembly by turning the Shaft Retaining Nut clockwise until the nut is tightened down on the compression fitting The paddle shaft should not be able to be withdrawn from the compression fitting The paddle assembly should spin freely on the shaft if pushed in a circular motion in any direction PROCULTURE SPINNER FLASKS OPERATING INSTRUCTIONS Drive Shaft Assembly y Housing Step1 Drive Shaft Lock Washer Step 3 a Coupli oupling Set Screws Collar Step 4 Paddle Assembly Figure 12 Direct Drive Cap Assembly 13 Cap Assemblies Direct Drive ProCulture Spinner Flasks 4510 and 4512 Series Flasks Connection of Direct Drive Systems with 100 mm or 120 mm Cap Assembly Fig 12 Step 1 Insert the ProCulture 4520 104 4520 106 Series Drive Shaft Assembly Housing into the center of the appropriate cap The flats on the threaded portion of the bottom of the drive help to orient the drive shaft in the cap The drive shaft should protrude out of the bottom of the cap assembly Step 2 Tighten the Lock Washer onto the threads of the Direct Drive Housing under the cap Step 3 Slide the assem
25. dle also ensures adequate aeration of the cell culture medium at slower speeds High speed stirring can be detrimental and is best done by slowly adapting cells to the higher shear forces which often develop at the higher speeds Initial stirring speeds for flasks of various flask sizes are listed below These speeds should be regarded as suggestions only and final agitation rpms need to be determined empirically Flask Size rpm 100 mL to 1 000 mL 50 60 3 000 mL to 8 000 mL 40 50 15 000 mL to 36 000 mL 30 40 Note It is important in scaling up that rpm in one flask should always be equated to tip speed in the larger flask Since the dimensions of the paddle may change from one flask to another using rpm alone will often result in detrimental cell viability This is because a larger paddle will travel much further at a given rpm than a smaller one There are a number of magnetic stirrer platforms or direct drive units that are available in the marketplace which will allow controlled agitation at these speeds PROCULTURE SPINNER FLASKS OPERATING INSTRUCTIONS 20 Disassembly Care and Maintenance Disassembly To disassemble the vessel for cleaning remove the cap paddle assembly from the vessel Loosen the shaft retaining nut holding the paddle in the cap assembly remove the split rings and pull out the paddle shaft Take precautions when removing a glass shaft from the cap assembly in the event that it has seized insid
26. e blades slightly while the paddle assembly is being slid into the center neck of the vessel If the blades should pop free from the blade retaining ring lift the assembly up in the vessel and gently reinsert the blades back into the notches in the blade retaining ring Secure the screw cap onto the vessel by tightening it slightly on the threads of the center neck Important For the Shur Loc Cap Assemblies 70 mm 100 mm 120 mm Production Style 4500 series and 4502 series Vessels and 140 mm cap assemblies on 4504 series and 4514 series with 140mm center neck openings DO NOT expand the O ring on the cap Simply insert the assembled cap and paddle assembly into the flask allowing it to loosely rest on the neck of the flask The gasket will expand during autoclaving and seal the vessel automatically Important Do not tighten any of the cap assemblies Center neck or Sidearm on the vessel Over tightening or securing the cap too tightly in or on the vessel can result in implosion of the vessel during autoclaving and or fracturing of the center neck opening during autoclaving Loosely place the caps onto the sidearm ports A slight clockwise turn of the cap onto the glass is sufficient to keep the caps from falling off the vessel while transporting it to the autoclave The vessel is now ready for autoclaving Note Use of Vented Membrane Cap Corning Cat No 1395 45L TMC is recommended as it will allow gas to exchange
27. e cleaning PROCULTURE SPINNER FLASKS OPERATING INSTRUCTIONS solution as soon as cell culture is terminated Disassemble the vessels to facilitate penetration of the cleaning agent Several protocols are described below After these treatments have been performed be sure to wash all components again as was described early in this manual in the Standard Glassware Cleaning Protocol in the section Cleaning and Siliconizing Your Vessel on page 7 Glassware cleaning is reasonably straightforward Silicone and synthetic rubber components tend to absorb cleaners and other chemicals and so more caution must be taken to minimize contact with them Depyrogenation 1 Baking Glassware can be baked for at least 12 hours at 250 F 120 C then clean as described in the Standard Glassware Cleaning Protocol on page 7 DO NOT USE HEAT TREATMENT ON ANY OF THE PLASTIC OR SILICONE SYNTHETIC RUBBER COMPONENTS 2 Chemical Treatment Soak flask components in solutions of sodium hydroxide 0 1 N to 1 0 N NaOH for at least 1 hour followed by soaking and then repeated wash ing with distilled or deionized water Io ensure that all traces of NaOH are gone check the pH of the soaking solutions to ensure that they are not alkaline Some investigators chase the NaOH treatment with a weak solution 0 1 N to 1 0 N of phosphoric acid to neutralize the NaOH prior to rinsing in distilled water Removal of Biological and Organic Residues
28. e the cap compression fitting If the shaft does seize place the cap paddle assembly into an autoclave and heat for a few minutes to soften the plastic components This will usually allow the shaft to be removed without breaking As a precaution wrap the shaft in a towel as it is removed to prevent cuts in case it does snap After removal follow the assembly directions in reverse to finish disassem bling the paddle and cap Note We recommend placing the parts into a dilute cleaning solution to soak as soon as possible after disassembly If the flask needs to be decon taminated pour out the contents into a separate vessel kill tank and fill the flask including the internal components as soon as possible with a bleach solution After treatment rinse thoroughly and wash or soak in the cleaning solution as above Care and Maintenance The ProCulture vessels have been constructed of materials that should allow multiple autoclavings for extended periods of time if the flask is properly cared for Extended autoclaving times at elevated temperatures or soaking the vessel or parts in acids bases or strong oxidizing agents for unusually long lengths of time will result in these materials deteriorating more quickly than usual Basic Care Care should be taken when washing the vessels to minimize bumping against other glassware or the sides of sinks so that stress fractures do not develop in the glass and result in failure during autocl
29. eads This is generally suitable for operation with 1 4 inlet gassing fitting PROCULTURE SPINNER FLASKS OPERATING INSTRUCTIONS 24 Frequently Asked Questions and Troubleshooting Guide Use the following guide to help you with the occasional problem you may experience with your ProCulture Spinner Flasks These commonly asked questions will guide you prior to calling our Technical Support Department 1 800 492 1110 in the U S Outside the U S please call your local Corning Representative The medium in my flask is turning alkaline prior to addition of cells The flask may have been improperly washed using a detergent not designed for cell culture Rewash the flask and components using a detergent designed for cell culture and rinse thoroughly with distilled water See Cleaning and Siliconizing Your Vessel page 7 The impeller in my magnetically driven flask is not turning smoothly The flask may not be centered above the magnet inside the magnetic stirrer Iry moving the flask slightly on the magnetic stirrer until smooth motion is observed The impeller in my magnetically driven flask turns smoothly at slow speeds but begins to turn in a disruptive or intermittent manner as the speed of the magnetic stirrer is increased The speed of the magnetic stirrer is set too high or has been increased too quickly As a result magnetic decoupling is occurring Or the viscosity of the medium is too high also resulting in d
30. ecoupling What volume should I place in the flask For New Users It is good to start at 50 of the working volume of the flask In sub sequent runs add 10 more volume each time until cell density plateaus or maximum working volume of flasks is achieved For Users of Conventional Flasks One should start at the same volume as your current flask In subsequent runs add 10 more volume each time until cell density plateaus or maximum working volume of flasks is achieved Note If the cell density plateaus before the maximum working volume is achieved you may need to use an augmented operation to boost productivity What is the correct speed to spin a flask and how do I adjust the speed as my flask size changes First look at the recommended guidelines for each flask size on page 19 For New Users In small flasks keep tip speed between 3 5 in sec In large flasks keep tip speed between 10 15 in sec In subsequent runs try and increase flask tip speed until cell density plateaus or cell viability drops For Users of Conventional Flasks One should start at the same volume as your current flask In subsequent runs try and increase the flask tip speed until cell density plateaus or cell viability drops PROCULTURE SPINNER FLASKS OPERATING INSTRUCTIONS UI Why Direct Drives and what size The decision to choose a direct drive is mainly a function of the fact that magnetic decoupling occurs at greater
31. ents Chemical Compatibility Chart 34 Ordering Information accostata honda indiano 35 Replacement Parts for Spinner Flasks L LL 40 PROCULTURE SPINNER FLASKS OPERATING INSTRUCTIONS Technical Service and Safety Information Technical Service Call 800 492 1110 outside the U S 001 978 635 2200 Fax 1 978 635 2476 or contact your local support office Safety Information for ProCulture Glass Reservoirs Since glass reservoirs cannot be guaranteed for any positive or negative pressure evacuated or pressurized glass reservoirs must be shielded The ProCulture Spinner Flasks described in this instruction manual all include at least one glass reservoir Glass has traditionally been the material of choice for Spinner Flasks because it is relatively inexpensive easy to clean provides high visibility of reservoir contents and is autoclavable Many researchers sparge their vessels or provide gas overlays to improve oxygenation of the flask medium Even if there is no intent to pressurize a reservoir there is the chance of accidentally pressurizing a glass reservoir during gassing if the sidearm caps are tightened or a vent filter becomes blocked Special precautions should therefore be taken It is not possible for a manufacturer of glass reservoirs to guarantee that its reservoirs will hold any specific positive or negative pressure without breaking For this reason ProCulture Spinner Flasks should always be shielded i
32. eparately Drive shaft housing made of rugged Noryl with internal Kevlar composite bushings PTFE spring energized seals internal air space vent and internal stainless steel drive shaft Spring energized seals ensure that shaft is truly sealed from the external environment unlike other drive units Rugged high torque drive motors deliver a constant 14 5 in lbs of torque to a maximum speed of 350 rpm for easy stirring of viscous solution such as media with micro carrier beads 4515 Paddle Assemblies for Direct Drive Stirring Systems For 4510 and 4512 series flasks coupled to a Direct Drive Mechanism Paddle Assemblies will couple to 100 or 120 Cap and Drive Shaft Assemblies For ordering please go to our Ordering and Replacement Parts Lists at the end of this manual PROCULTURE SPINNER FLASKS OPERATING INSTRUCTIONS Flask Details Schematic Circuits 0 2 um Gas 4519 100 Filter l 4519 102 4519 106 k L00 00606e0 Flow Meter Compressed Gas or Gas Source Peristallic Pump Figure 31 Continuous Sparging Schematic 28 PROCULTURE SPINNER FLASKS OPERATING INSTRUCTIONS 29 Flask Details Schematic Circuits 4519 112 4519 116 or 4519 120 Overflow Phase Addition Phase Ta 4519 112 4519 116 or 4519 120 dI 4519 110 Figure 32 pH Control Schematic Diagram PROCULTURE SPINNER FLASKS OPERATING INSTRUCTIONS 30 Flask Detail
33. ession Lower Plate with Flats 140 mm Cap Compression Upper Plate with Flats 140 mm Cap Seal Actuating Ring 140 mm Cap Washer Seal Actuating Ring 140 mm Cap O ring Retaining Size 123 140 mm Cap Gasket Compression Sealing Size 428 Nut Shaft Retaining BP 3413 9041 Nut Shaft Retaining for 1 4 O D 6 mm Shafts ULTEM BP 3413 9042 Nut Shaft Retaining for 5 16 O D 8 mm Shafts BP 3812 005 ULTEM O ring for Shaft Retaining Nut Direct Drive Cap Lock Washer BP 3921 005 Direct Drive Cap Compression Fitting Lock Washer For ordering and technical service of all BP 3xxx xxx part numbers please call Wilmad LabGlass at 1 800 220 5171 ext 686 in the U S Outside the U S please call 001 856 697 3000 ext 686 For all other part numbers please call Corning Incorporated at 1 800 492 1110 in the U S Outside the U S please call your local Corning Representative 41 PROCULTURE SPINNER FLASKS OPERATING INSTRUCTIONS Replacement Parts for Spinner Flasks Replacement Parts for Spinner Flask Sidearm Fittings PET Inserts BP 3802 001 BP 3803 001 BP 3807 001 BP 3812 001 BP 3822 001 BP 3827 001 BP 3832 001 BP 3832 002 45 mm PET Insert for Single 1 8 Inlet 45 mm PET Insert for Single 1 4 Inlet 45 mm PET Insert for 50 mm Filter 45 mm PET Insert for 12 mm Insertion Probes 45 mm PET Insert for Dual 1 8 Media Handling Sidearm Fitting 45 mm PET Insert for Dual 1 4 Media Handling Sidearm F
34. f gassing of the vessel is attempted In addition good laboratory procedure should always be followed in handling and cleaning glassware The use of personal eye protection should be routine See below for a full statement of limitation of liability regarding ProCulture glass reservoirs Caution being of individuals using these products is dependent on their proper operation maintenance and safe laboratory practice Refer to the owners guide product package inserts and labels concerning warnings safe use and operation of any Corning laboratory product q Corning laboratory glass is intended for laboratory use only The well 4 PROCULTURE SPINNER FLASKS OPERATING INSTRUCTIONS Introduction Welcome Note For optimum performance and safety please read these instructions carefully Thank you for choosing the ProCulture Spinner Flask System By investing in this system we feel that you will get Increased efficiency Ease of use Better performance with many cell lines State of the art cell culture capabilities Safety Instructions The following precautions should be followed any time you work with glass vessels Safety Glasses should be worn at all times Take extreme caution whenever you handle glassware especially when washing and assembling disassembling spinner flasks The most damage and injuries may occur during this time We strongly recommend that the flask is replaced if any cracks or chips appear a
35. front Plaza Corning NY 14831 0001 United Kingdom t 0800 376 8660 f 0800 279 1117 LATIN AMERICA Brasil t 55 11 3089 7419 f 55 11 3167 0700 Mexico t 52 81 8158 8400 f 52 81 8313 8589 Printed in USA 1 06 POD CLS BP 018 REVI 2006 Corning Incorporated
36. g Ring PET Yes Yes Yes Yes Washer Paddle Assemblies Bearing Holder PTFE Yes Yes Yes Yes Paddle Blade PTFE Yes Yes Yes Yes Magnet Encased PTFE Yes Yes Yes Yes Stainless Steel Shaft 316L Yes Yes Yes Yes Glass Shaft Corning 7740 Type I Borosilicate Yes Yes Yes Yes Plastic Shafts Polysulfone Yes Yes Yes Yes Bearing Media Plug Silicone Yes Yes Yes Yes Bearing Thrust Washer PTFE Yes Yes Yes Yes Blade Retaining Sleeve PTFE Yes Yes Yes Yes Direct Drive Assembly Thrust Washer Bearings Torlon 4301 Yes Yes Yes Yes Shaft O ring Viton Yes Yes Yes Yes Shaft Housing Noryl Yes Yes Yes Yes Metal Components 316 Stainless Steel Yes Yes Yes Yes Vent Fitting Polypropylene Yes Yes Yes Yes Shaft Seal Housing Hydlar ZF Yes Yes Yes Yes Shaft Seals Teflon Proprietary Yes Yes Yes Yes Cap Lock Nut PET Yes Yes Yes Yes Snap Ring 316 Stainless Steel Yes Yes Yes Yes Product Chemical Composition Viton Fluoroelastomer Noryl Polyphenylene Oxide EN 265 PET Polyethylene Terephthalate Hydlar ZF Kevlar Nylon Teflon PTFE Torlon 4301 Polyamide imide Valox 420 PBT 30 glass ULTEM 1000 Polyetherimide PBT Polybutylene Terephthalate For more detailed information on chemical resistivity please contact the manufacturer 34 PROCULTURE SPINNER FLASKS OPERATING INSTRUCTIONS Ordering Information Corning Life Sciences is currently transitioning ProCulture Spinner Flask s
37. ildup of cells or microcarriers on the surface of the glassware is noted The protocols described above are general procedures which work well with glassware that has had little use In the event that the vessel has been used extensively and is markedly soiled please refer to the section entitled Disassembly Care and Maintenance page 20 prior to siliconizing the glassware PROCULTURE SPINNER FLASKS OPERATING INSTRUCTIONS Assembly of Vessels and Components Paddle Assemblies Magnetically Driven ProCulture Spinner Flasks 4500 4502 and 4504 Series Flasks All paddles for vessels which are magnetically driven 4500 series 4502 series and 4504 series flasks are assembled in essentially the same manner as described below b fam PTFE Bearing Holder Step 2 Insert shaft into Bearing Holder Set aside PTFE Split Ring PolySulfone or Stainless Steel Shaft PTFE Split Ring Step 1 Step 3 Insert Silicone Plug optional Silicone bo a PTFE Bearing Holder Step 2 Insert shaft into Bearing Holder Step1 Set aside PTFE Split Ring Glass Shaft Step 3 Insert Silicone Plug optional Silicone Plug J c Y Blade Retaining Step 5 Sleeve PTFE Silicone Plug Bearing Holder Hole for the Magnet Step 4 Paddle Blade Hole for the Magnet O d Glass PolySufone or Stainless Steel Shaft Blade Retaining Sleeve Step 5 Bearing Holder continued
38. ing vent cap 0 2 pm 50 mm filter l 4519 177 Sidearm fitting vent cap 0 2 pm Sanitary 50 mm filter l Media Handling Fittings Vertical Sidearm Flasks Used to introduce medium aseptically into large spinner flasks with verti cal sidearms polypropylene sealing cap Gas filters are PTFE 0 2 pm porosity grated ferrules The fittings are completely autoclavable Media Handling Fittings Vertical Sidearm Flasks Ordering Information Fittings are comprised of a PET insert with a Viton O Ring and a The 316 stainless steel tubes are held in place by Noryl nuts with inte Fits Tubing O D Qty Cat No Description Flask Size inches Case 4519 112 Sidearm fitting dual media handling 8L 15L 1 8 1 4519 114 Sidearm fitting dual media handling 36L Vg 1 4519 116 Sidearm fitting dual media handling SL 15L 1 4 1 4519 118 Sidearm fitting dual media handling 36L 1 4 1 4519 120 Sidearm fitting combo media handling 8L 15L 1 8 1 4 1 4519 122 Sidearm fitting combo media handling 36L 1 8 1 4 1 4519 124 Sidearm fitting single media handling 8L 15L 1 2 I 4519 126 Sidearm fitting single media handling 36L 1 2 I 4519 176 Sidearm fitting dual media handling SL 15L 1 4 1 EPDM PROCULTURE SPINNER FLASKS OPERATING INSTRUCTIONS 37 AO OS Dual Angled Sidearm Fittings Combination and Triple Angled Sidearm Fittings Gas or Media Handling Fittings Angled Sidearm Flasks Dual Style
39. itting 45 mm PET Insert for Combination 1 8 1 4 Media Handling Sidearm Fitting 45 mm PET Insert for Triple 1 8 Inlet Stainless Steel Tubing BP 3802 007 BP 3803 007 BP 3822 007 BP 3827 007 316 Stainless Steel Tubing 1 8 O D x 18 For 8L and 15L Flasks 316 Stainless Steel Tubing 1 4 O D Threaded x 18 For 8L and 15L Flasks 316 Stainless Steel Tubing 1 8 O D x 21 For 36L Flasks 316 Stainless Steel Tubing 1 4 O D x 21 For 36L Flasks Angled Sidearm Replacement Tubing Please Contact Corning Customer Service Compression Nuts amp Ferrules BP 3802 004 BP 3803 004 BP 3812 004 BP 3847 008 BP 3852 008 BP 3857 008 1 8 Nut Noryl 1 4 Nut Noryl 12 4 mm Nut Noryl 1 8 Plug Delrin 1 4 Plug Delrin 12 mm 1 2 Plug Delrin Caps O rings Filters and Caps BP 3802 002 BP 3802 003 BP 3802 006 BP 3807 004 BP 3812 005 1395 4SLIMC Insert O ring Viton 45 mm Securing Cap Bored Stainless Steel Sparging Stone amp Nut Complete 10 micron Mesh 50 mm PTFE Filter 0 2 micron NPT ends O ring for 12 4 mm Nut Corning 45 mm Low Temperature Vent Cap with 0 22 micron Filter Securing Caps BP 3802 003 BP 3872 002 1395 32LTC 1395 45LTC 1395 45LTR 1395 45LTMC i Cap securing 45 mm bored orange Cap securing 45 mm bored for injection septum white Cap solid 32 mm orange Cap solid 45 mm orange Drip ring 45 mm clear Cap vented
40. k or Sidearm Introduction 1 Open the vessel after it cools by loosening the center cap assembly and gently shifting the cap and paddle slightly over allowing it to rest on the lip of the center neck opening or remove sidearm cap and rest in hood 2 Introduce medium by pouring from pre sterilized bottles or bags con taining medium by gravity feed or peristaltic pumps as shown in Fig 29 3 In line addition is an alternative Unsterilized medium can be added by filtering in line using a length of sterilized tubing connected to a 0 2 micron filter attached between the unsterilized medium source and the vessel Either gravity feed or a peristaltic pump can be used to move the fluid By using a filling bell supported by a ring stand as shown in Fig 30 several vessels can be rapidly filled Large vessels Sidearm Introduction 1 Medium addition is usually done using the in line sterilization method described above 2 With larger flasks medium addition is usually done via the sidearms rather than the center neck Again the use of a filling bell supported by a ring stand as shown in Fig 30 greatly simplifies this operation 0 22 micron In line Filter Q e 20 Peristaltic Pump Unfiltered Medium Figure 30 Large vessel being filled with the use of a filling bell and ring stand PROCULTURE SPINNER FLASKS OPERATING INSTRUCTIONS Inoculating and Agitating the Vessel Inocu
41. l damages from economic loss or property damage sustained by any Purchaser from the use of CORNING products This limited warranty excludes coverage on glass components and probes including cables CORNING reserves the right to make substitutions and modifications in the design and or specifications of products set forth in sales literature providing that substitutions or modifications do not materially and adversely affect the performance of the product 2 PROCULTURE SPINNER FLASKS OPERATING INSTRUCTIONS Contents Technical Service and Safety Information 4 INFEOGUGHOD 24524025 diante ao 5 MCO eraran EEEE PAE E EE E EO 5 Safety IMSWUCHONS ssi a A 5 Checking Your Model Number 5 Getting Started oss i uasavadnbpanisog idres tradidisti enikeni 6 Cace eree ees a ideare 6 Cleaning and Siliconizing Your Vessel 0 00 00 02 eee ee 7 Assembly of Vessels and Components L LL 8 Final Assembly of Vessels and Autoclaving 04 17 Basic OMCIAUOO 2 46 62 inalterate 18 Filling the Vessel with Medium LL 18 Inoculating and Agitating the Vessel 00005 19 Disassembly Care and Maintenance 0 0 eee ee eee 20 Additional Information LL 22 Applications Check Lit resina tarata ra 22 Frequently Asked Questions and Troubleshooting Guide 24 Optional Operation 0 cece eee eee eens 26 Plas Weal cinisello 28 Spinner Flask Compon
42. lating Inoculating the vessel can be accomplished in a variety of ways and is best done in a sterile biological hood or a clean room After the vessel passes the sterility check cells can then be harvested and safely added to the vessel Cells can be introduced by reconstituting cell pellets in a smaller volume of medium after harvesting Adding cells is usually done using a syringe or length of sterile tubing into the center neck opening or sidearms Inoculation densities are usually determined empiri cally using smaller test vessels such as T flasks or small spinner flasks For cells which are anchorage dependent sterile microcarrier beads can be added to the vessel prior to or during the inoculation of the vessel Cells and beads are often agitated intermittently during the attachment phase of the cells to the beads A good reference text such as Culture of Animal Cells A Manual of Basic Technique by R Ian Freshney Alan R Liss Publishing New York 1994 or Cell and Tissue Culture Laboratory Procedures by Doyle A et al John Wiley amp Sons Sussex England 1995 or our Technical Service Department 800 492 1110 or outside the U S 001 978 635 2200 can help you with these procedures Agitating The ProCulture Spinner Flasks have been engineered to achieve excellent mixing of vessel contents using slow speeds The combination of the paddle design and vessel baffles provide optimal uplift The surface breaking design of the pad
43. lug ED Figure 5 Direct Drive Paddle Assembly 10 Paddle Assemblies Direct Drive ProCulture Spinner Flasks 4510 4512 Series Flasks Direct drive paddles 4515 are assembled as shown in Fig 5 Step 1 Slide the Paddle Blade into the notch in the Bearing Holder narrow end of the paddle blade first until the hole on the blade lines up with the hole in the bearing holder Step 2 Insert the short rounded Retaining Plug into the aligned holes until it is centered in the combined assembly The Paddle Blade should not be able to slide on the bearing holder Step 3 Take the Blade Retaining Sleeve with the notched end facing down and slide it down on top of the newly assembled Bearing Holder and Paddle Blade Position the narrow ends of the paddle blade into the notches of the Blade Retaining Sleeve and push down until the top portions of the blade extend completely into the notch in the retaining sleeve Step 4 Slide down one end of the Stainless Steel Coupling Collar onto the Stainless Steel Shaft The other part of the coupling collar will slide up onto the flat portion of the drive unit shaft in a later assembly For more detail see Figures 6 through 9 Step 5 Tighten the set screws on both sides of the collar so that the collar does not turn on the shaft Figure 6 Coupling collar larger opening for paddle shaft Figure 7 Coupling collar smaller opening for drive shaft Figure 8 Slide l
44. m O D Paddle Glass Rods 36000 mL 8 mm O D Paddle Rods Stainless Steel Dual Bearing BP 3611 081 BP 3611 081 BP 3611 082 BP 3611 083 BP 3611 084 BP 3611 085 BP 3611 086 BP 3611 087 BP 3611 088 Paddle Stainless Steel Rods 125 mL 1 4 O D Paddle Stainless Steel Rods 250 mL 1 4 O D Paddle Stainless Steel Rods 500 mL 1 4 O D Paddle Stainless Steel Rods 1000 mL 1 4 O D Paddle Stainless Steel Rods 3000 mL 5 16 O D Paddle Stainless Steel Rods 6000 mL 5 16 O D Paddle Stainless Steel Rods 8000 mL 5 16 O D Paddle Stainless Steel Rods 15000 mL 5 16 O D Paddle Stainless Steel Rods 36000 mL 5 16 O D Paddle Silicon Plugs BP 3411 090 BP 3411 095 Paddle Silicon Plugs 125 mL 1000 mL Paddle Silicon Plugs 3000 mL 36000 mL Direct Drive Bearing Holders BP 3711 016 BP 3711 017 BP 3711 018 Direct Drive Paddle Blade Holder with Stainless Steel Shaft 8000 mL Direct Drive Paddle Blade Holder with Stainless Steel Shaft 15000 mL Direct Drive Paddle Blade Holder with Stainless Steel Shaft 36000 mL Direct Drive Paddle Retaining Plug BP 3711 066 Direct Drive Paddle Retaining Plug Direct Drive Coupling Collar BP 3711 096 BP 3711 094 Stainless Steel Coupling Collar Screw Set For Coupling Collar 1 4 20 X 1 4 316 Stainless Steel Direct Drive Paddle Blades BP 3711 026 BP 3711 027 BP 3711 028 Paddle Blades 8000 mL Paddle Blades 15000 mL Paddle Blades 36000 mL Cap Assemblies Co
45. mplete BP 3402 100 BP 3403 100 BP 3404 100 BP 3404 100 BP 3602 100 BP 3504 106 BP 3504 112 70 mm Cap Assembly Magnetic style Complete for 125 mL 250 mL Flasks 100 mm Cap Assembly Magnetic style Complete 1 4 6 mm for 500 mL 1000 mL Flasks 100 mm Cap Assembly Magnetic style Complete 5 16 8 mm for 3L 36L Flasks 120 mm Cap Assembly Magnetic style Complete for 3L 36L Flasks 140 mm Cap Assembly Magnetic style Complete for 3L 36L Flasks 120 mm Cap Assembly Magnetic style Complete with 2 12 mm Fittings 120 mm Cap Assembly Magnetic style Complete with 2 12 mm and 2 1 4 Fittings Cap Assembly Components Center Neck Caps BP 3412 011 BP 3412 015 BP 3413 011 BP 3413 015 BP 3516 011 BP 3513 015 BP 3518 012 BP 3513 016 BP 3518 015 BP 3514 016 BP 3612 010 BP 3612 020 BP 3612 030 BP 3612 050 BP 3612 060 BP 3612 070 70 mm Cap Screw Type with Integrated Compression Fitting ICF Liner 70 mm with Center Hole Red Silicone 100 mm Cap Screw Type with ICF Liner 100 mm with Center Hole Red Silicone 120 mm Cap Screw Type with ICF Liner 120 mm with Center Hole Red Silicone 120 mm Cap Screw Type with ICF with 2 12 mm Holes Liner 120 mm with Center Hole Red Silicone with 2 12 mm Holes 120 mm Cap Screw Type with ICF with 2 12 mm 2 1 4 Holes Liner 120 mm with Center Hole Red Silicone with 2 12 mm 2 1 4 Holes 140 mm Cap Compr
46. n Qty Case 402579 Cap Assembly 120 mm Glass Filled PBT 3 3 8 1 1 1 4 fittings 402576 Cap Assembly 120 mm Glass Filled PBT 2 12 mm 1 2 4 4 fittings 402577 Cap Assembly 120 mm Glass Filled PBT 2 12 mm 1 2 1 4 1 3 8 fittings Direct Drive Motors High torque low rpm stirrer designed to maintain constant low speed Gearhead stirrer delivers 14 5 in lbs of torque Maximum speed is 350 rpm Weight of motor is 9 Ibs 4 1kg Available with 120VAC 60Hz or 230VAC 50Hz Direct Drive Motors Ordering Information Cat No Description Qty Case 400640 120VAC 60 Hz Motor l 402645 230VAC 50 Hz Motor l Direct Drive Shaft Cap Assemblies For 8L 15L or 36L paddle assemblies Used on all series 4510 and 4512 Spinner flasks Direct Drive Shaft Cap Assemblies Ordering Information Cat No Description Qty Case 402614 For 100 mm Neck Flasks 1 402649 For 120 mm Neck Flasks 1 Direct Drive Paddle Assemblies For series 4510 4512 and 4514 Spinner flasks when coupled to a direct drive motor Paddle assemblies will couple to 100 mm and 120 mm cap assemblies Direct Drive Paddle Assemblies Ordering Information Cat No Description Qty Case 4515 8L Paddle assembly only for 8L flask 1 4515 15L Paddle assembly only for 15L flask l 4515 36L Paddle assembly only for 36L flask 1 PROCULTURE SPINNER FLASKS OPERATING INSTRUCTIONS Replacement Parts for Spinner Flasks Repla
47. nd poor agitation Therefore be sure to match the correct fitting and shaft to eliminate any problems Step 1 Figure 10c Secure the Silicone Liner into the top of the Screw Cap with Integrated Compression Fitting In order to help secure the Silicone Liner the integrated compression fitting and outer edge of the screw cap have been undercut to create a notch into which the liner fits Step 2 Figure 10a Remove the Shaft Retaining Nut from the cap Slide the knurled larger end of the Shaft Retaining Nut onto the glass polysulfone or stainless steel Impeller Shaft on the Assembled Paddle Step 3 Figure 10b Insert the PTFE Split Ring back onto the notch of the shaft from which it was previously removed during the Paddle Assembly process page 8 Step 4 Figure 10c Insert the Completed Paddle Assembly and Retaining Shaft into the Screw Cap with Integrated Compression Fitting and tighten the assembly by turning the Shaft Retaining Nut clockwise until the nut is tightened down on the com pression fitting The paddle shaft should not be able to be withdrawn from the compression fitting The paddle assembly should spin freely on the shaft if pushed in a circular motion in any direction Figure 10 Magnetically Driven Screw Cap Assembly 11 PROCULTURE SPINNER FLASKS OPERATING INSTRUCTIONS SS Retaining O Ring I Seal Actuating Ring Washer fammi Seal Actuating me Ring Step 5 Step 3 Upper Plate
48. ns that are exposed on the motor drive adapter to help you orient the inside portion of the dowel pins with the bayonet fitting as shown in Fig 24 You will know that the dowel pins are corrently oriented when the motor unit drops slightly into the vertical slots Step 5 Figure 25 When the dowel pins have dropped into the slots turn the motor slightly clockwise to lock the entire assembly together as shown in Fig 25 Figure 21 Engagemnet slots for bayonet Figure 22 Closeup of bayonet fittings fittings and yoke dowel pins ni p e Figure 23 Coupling of motor and Direct Figure 24 Correct orientation of motor Drive Shaft on Drive Shaft Figure 25 Assembly is locked in place PROCULTURE SPINNER FLASKS OPERATING INSTRUCTIONS T 0 Pad er Shaft To PI M Assembly z 7 e Step 1 ne Drive Shaft Bottom Plate Bellco U O Drive Only Step 2 Screw Drive Housing into Cap Assembly a ni Step 3 Slide Collar on to Drive Shaft a a and Paddle step 4 Shaft Tighten Set SCIEWS Assembled Paddle Figure 26 ProCulture 100 mm 120 mm Cap with a Bellco Direct Drive System Cap Assemblies Direct Drive ProCulture Spinner Flasks 4510 and 4512 Series Flasks continued Connection of Bellco Technology Direct Drive Systems with a ProCulture 100 mm or 120 mm Cap Assembly Fig 26 Step 1 If using a Bellco OverHead Drive remove the bottom plate from the Bellco OverHead Drive Cap
49. nywhere on the vessel center neck cap or sidearms This will help to minimize injury since flasks can develop micro fractures which can easily cause catastrophic failure during autoclav ing or handling Under no circumstances should the vessel be pressurized or evacuated without precautions For more information see page 4 Checking Your Model Number This manual provides instructions for assembly and operation of the following ProCulture Spinner Flasks 4500 4502 4504 4510 4512 4514 series There are 6 basic styles of Corning Spinner Flasks shown below 4500 series 70 mm amp 100 mm Center Neck Cap Angled Sidearms 4502 series 120 mm Center Neck Cap Angled Sidearms 4504 series 140 mm Center Neck Cap Angled Sidearms 4510 100 mm 4512 120 mm 4514 140 mm series Center Neck Cap Vertical Sidearms The flask style can be determined by comparing it to those shown on the next page Any differences in assembly or operation is clearly marked in the text PROCULTURE SPINNER FLASKS OPERATING INSTRUCTIONS Getting Started Figure 1a Cat No 4500 4502 4504 Series Figure 1b Cat No 4510 4512 4514 Series Unpacking Note Due to variations in flask styles some components may be intentionally not included as indicated below Corning Spinner Flasks 4500 Series and 4502 Series Packaged with Cap Style a Cap Assembly Screw Cap Fig 2a Cap Style b Shurloc Cap Assembly Compressi
50. o the bottom of the flask the other is a shorter 6 length Both tubes are held in place by Noryl nuts with integrated ferrules The fittings are completely autoclavable Gas or Media Handling Fittings Angled Sidearm Flasks Combination Style Ordering Information Flask Tubing O D Qty Cat No Description Size inches Case 4519 158 Sidearm fitting combination IL 1 8 1 4519 159 Sidearm fitting combination 3L 1 8 4519 160 Sidearm fitting combination 6L 1 8 l 4519 161 Sidearm fitting combination 8L 1 8 4519 162 Sidearm fitting combination IL 1 4 4519 163 Sidearm fitting combination 3L 1 4 4519 164 Sidearm fitting combination 6L 1 4 l 4519 165 Sidearm fitting combination 8L 1 4 4519 171 Sidearm fitting combination I5L 1 4 4519 166 Sidearm fitting combination triple IL 1 8 1 4519 167 Sidearm fitting combination triple 3L 1 8 1 4519 168 Sidearm fitting combination triple 6L 1 8 4519 169 Sidearm fitting combination triple 8L 1 8 PROCULTURE SPINNER FLASKS OPERATING INSTRUCTIONS 38 s m n nur 18 mm 12mm Sidearm Fittings for Sensors Impeller Assembly Fittings for Insertion Probes Vertical Sidearm Flasks Used to secure pH O or temperature sensors in large spinner flasks with vertical sidearms Fittings are comprised of a PET insert with a Viton O ring and a polypropylene sealing cap The 316 sensors are hel
51. on Cap Fig 2b is available as an option Packaged with Magnetically Driven Paddle with Polysulfone or Glass Shaft Fig 3a 4504 Series Packaged with Cap Style b Shur Loc Cap Assembly Compression Cap Fig 2b Packaged with Magnetically Driven Paddle with Glass Polysulfone or optional Stainless Steel Shaft Fig 3a 4510 4512 4514 Series Packaged as flask only Cap and Direct Drive Paddle Assemblies Fig 3b must be purchased separately No spare parts are included Figure 2a Style a Cap Assembly Figure 2b Style b Shur Loc Cap Screw Cap Assembly Compression Cap Figure 3a Magnetically Driven Paddle Figure 3b Direct Drive Paddle Assembly with Glass Polysulfone or optional Stainless Steel Shaft PROCULTURE SPINNER FLASKS OPERATING INSTRUCTIONS Cleaning and Siliconizing Your Vessel Standard Glassware Cleaning Protocol Although your vessel and parts were extensively cleaned prior to packing we strongly recommend that you wash the vessel and components again to remove any residue that may have developed in the vessel after packing It is easiest to do this prior to assembling the flask There are several deter gents on the market that are available for this purpose including 7X Original Cleaning Solution and Sigmaclean that can be purchased from your local lab supplier Do not use detergents such as Alconox or others that were developed for chemistry applications since these cleaner
52. pare parts to Wilmad Lab Glass For ordering and technical service of all products that have BP xxxx xxx part numbers please call 1 800 220 5171 ext 686 in the U S Outside the U S please call 001 856 697 3000 ext 686 For all other part numbers that are not associated with a BP xxxx xxx part number please call Corning Incorporated at 1 800 492 1110 in the U S Outside the U S please call your local Corning Representative ProCulture Glass Spinner Flask with Angled Sidearms Baffles enhance aeration and agitation of contents of the flask Unique impeller design ensures optimal stirring Sidearm designs permit easy access of 25 and 50 mL pipettes Visit www corning com lifesciences to view additional Corning spinner flask accessories ProCulture Spinner Flasks with Angled Sidearms Ordering Information Center Sidearm Qty Cat No Description Capacity Neck mm Neck mm Case 4500 125 Spinner 125 mL 70 32 l 4500 1L and 4500 250 4500 250 Spinner 250 mL 70 32 1 Spinner Flasks 4500 500 Spinner 500 mL 100 45 1 4500 1L Spinner IL 100 45 l 4500 3L Spinner 3L 100 45 l 4500 6L Spinner 6L 100 45 l 4500 8L Spinner 8L 100 45 l 4500 15L Spinner ISL 100 45 l 4500 36L Spinner 36L 100 45 1 4502 3L Spinner 3L 120 45 l 4502 6L Spinner 6L 120 45 l 4502 8L Spinner 8L 120 45 I 4502 15L Spinner ISL 120 45 l 4502 36L Spinner 36L 120 45 1 4504 3L Spinner 3L 140 45 l 4504 6L Spinner 6L 140 45 l ProCulture S
53. pinner Flasks 4504 8L Spinner SL 140 45 1 4504 15L Spinner ISL 140 45 l 4504 36L Spinner 36L 140 45 l Retrofit Kits are available for converting older Corning ProCulture Spinner Flasks to fit newer dual bearing impellers ProCulture Spinner Flasks with Vertical Sidearms Ordering Information Center Number of Sidearm Qty Cat No Capacity Neck mm Vertical Sidearms Neck mm Case 4510 8L 8L 100 4 45 1 4510 15L ISL 100 4 45 1 4510 36L 36L 100 6 45 1 4512 8L 8L 120 4 45 1 4512 15L ISL 120 4 45 1 4512 36L 36L 120 6 45 1 4514 15L ISL 140 4 45 1 4514 36L 36L 140 6 45 1 PROCULTURE SPINNER FLASKS OPERATING INSTRUCTIONS 36 Vertical Sidearm Fittings Gas Delivery and Venting Vertical Sidearm Fittings Media Handling Gas Handling Fittings Vertical Sidearm Flasks Used to provide gases into larger spinner flasks with vertical sidearms Fittings are comprised of a PET insert with a Viton O Ring and a polypropylene sealing cap Gas filters are PIFE 0 2 pm porosity integrated ferrules The fittings are completely autoclavable Gas Handling Fittings Vertical Sidearm Flasks Ordering Information The 316 stainless steel tubes are held in place by Noryl nuts with Cat No Description Dimension Qty Case 4519 100 Sidearm fitting gas delivery 1 8 Inlet l 4519 102 Sidearm fitting gas delivery 1 4 Inlet 1 4519 104 Sidearm fitting delivery and vent 1 8 and 1 4 I 4519 106 Sidearm fitt
54. ries you will need please see our ordering section in the back of this manual A few summarized schematic circuit examples can be found on pages 28 29 that may meet your unit operation requirements For any additional help please contact your local Corning Application Specialist who can assist you further with any of the parts that you may need 4519 Accessories for ProCulture Spinner Flasks All fittings are made of rugged PET with compression fittings and ferrules made of Noryl All fittings are designed to fit standard 45mm GL 45 screw threads on ProCulture vessels Securing caps are made of native polypropylene Vent cap 4519 108 screw threads will accept most standard 50 mm 0 2 micron PTFE disposable filters Media handling and gas delivery tubes are made of 316L stainless steel Sensor fittings will accept most 12 mm insertion probes O ring made of Viton For ordering and selection of any sidearm fittings please see the Ordering and Spare Parts list in the back of this manual PROCULTURE SPINNER FLASKS OPERATING INSTRUCTIONS Direct Drive Stirring Systems and Cap Drive Shaft Assemblies for Baffled Flasks Agitation of the contents of Spinner Flasks at volumes greater than 8L can often become problematic using magnetic coupling Drag created by the paddle blade can overcome the coupling of the magnet in the flask with the magnet in the magnetic stirrer below the flask This effect becomes more p
55. ronounced as the volume in the flask increases Although magnetic cou pling may be sufficient to maintain the agitation in many cases Corning offers an Overhead Direct Drive System for the many instances where higher agitation rates greater volumes or higher viscosity or density cell culture such as microcarrier bead applications The direct drive system can be used with angled sidearm flasks but has its greatest utility with vertical sidearm flasks The design of the vertical sidearm flasks allows the insertion of probes and tubes to more easily provide the sophisticated bioreactor operations that were mentioned earlier by moving these components to the periphery of the flask allowing the center neck cap to be used for the drive shaft The optional larger 120mm center neck opening makes setup of these flasks even easier There are three components that are required for the use of the Direct Drive System Angled or Vertical Sidearm Flask Drive Motor Center Neck Cap with Drive Shaft Assembly Center neck cap and drive shaft assemblies will accept either 8000 15000 or 36000 mL paddle assemblies and will mate with ProCulture direct drive motors Optional cap assemblies are available to mate with other overhead drive motors Contact Corning for more information Drive shaft assembly and motors can be used on all 4510 and 4512 vertical sidearm flasks Cap assemblies DO NOT INCLUDE direct drive paddles which must be purchased s
56. rs please call Corning Incorporated at 1 800 492 1110 in the U S Outside the U S please call your local Corning Representative 42 PROCULTURE SPINNER FLASKS OPERATING INSTRUCTIONS Corning Incorporated Life Sciences 45 Nagog Park Acton MA 01720 t 800 492 1110 t 978 635 2200 f 978 635 2476 www corning com lifesciences For more technical or product information please refer to product literature and protocols Alternatively you may call Technical Services at 800 492 1119 or visit www corning com lifesciences Worldwide Support Offices ASIA PACIFIC Australia t 61 2 9416 0492 f 61 2 9416 0493 China t 86 21 3222 4666 f 86 21 6288 1575 Hong Kong t 852 2807 2723 f 852 2807 2152 India t 9111 341 3440 f 9111 3411520 Japan t 81 0 3 3586 1996 1997 f 81 0 3 3586 1291 1292 Korea t 82 2 796 9500 f 82 2 796 9300 Singapore t 65 6733 6511 f 65 6861 2913 Corning is a registered trademark of Corning Incorporated Corning NY Discovering Beyond Imagination and Flame of Discovery design are trademarks of Corning Incorporated Corning NY Taiwan t 886 2 2716 0338 f 886 2 2716 0339 EUROPE France t 0800 916 882 f 0800 918 636 Germany t 0800 101 1153 f 0800 101 2427 The Netherlands amp All Other European Countries t 31 0 20 659 60 51 f 31 0 20 659 76 73 All other trademarks included in this document are the property of their respective owners Corning Incorporated One River
57. rts PROCULTURE SPINNER FLASKS OPERATING INSTRUCTIONS Flask Details ProCulture 4504 Series Flask Spinner 140 mm Center Neck Magnetic Complete See Figure 34 Flask with 140 mm Center Neck Opening Complete Part Description BP 3605 xxx Replacement Flask with 140mm Center Neck Opening BP 3602 xxx 140mm Cap Assembly Magnetic style Complete A 140 mm Cap Assembly Magnetic style Complete B 140 mm Cap Compression Upper Plate C 140 mm Cap Compression Lower Plate D 140 mm Cap Seal Actuating Ring E 140 mm Shaft Retaining Nut F 140 mm Cap Washer Seal Actuating Ring G 140 mm Cap O ring Retaining Size 123 H 140 mm Cap Gasket Compression Sealing Size 428 BP 3401 xxx Impeller with Glass Shaft Magnetic Complete I 140 mm Paddle Bearing Holder J 140 mm Paddle Blade K 140 mm Paddle Glass Rod L 140 mm Paddle s s Rod Optional M 140 mm Paddle Retaining Sleeve N 140 mm Paddle Magnet O 140 mm Paddle Silicone Plug P PTFE Split Ring 1 per Glass Rod 2 per s s Rod B P s es IND an tz ga da K Or L Po da E Fd CORNING S N s15 L LAII Sal e I Paso N N eran Figure 34 ProCulture 4504 Series Flask Spinner 140 mm Center Neck Magnetic Complete PROCULTURE SPINNER FLASKS OPERATING INSTRUCTIONS 32 Flask Details ProCulture Direct Drive Detail with 100 120 mm Cap Assembly See Figure 35 4510 amp 4512 Serie
58. s ProCulture 4500 amp 4502 Series Flask Parts See Figure 33 4500 Series Flask Spinner 70 100 mm Center Neck Magnetic Complete 4502 Series Flask Spinner 120 mm Center Neck Magnetic Complete Part Description BP 3405 xxx Replacement Flask with 70 100 mm Center Neck Opening BP 3505 xxx Replacement Flask with 120 mm Center Neck Opening A Replacement Flask with 70 100 mm Center Neck Opening A Replacement Flask with 120 mm Center Neck Opening BP 3402 100 70 mm Cap Assembly Magnetic style Complete for 125 mL 250 mL Flasks BP 3403 100 100 mm Cap Assembly Magnetic style Complete for 500 mL 1000 mL Flasks BP 3404 100 100 mm Cap Assembly Magnetic style Complete for 3000 mL 36000 mL Flasks BP 3502 100 120 mm Cap Assembly Magnetic style Complete for 3000 mL 36000 mL B 70 mm Cap Screw Type B 100 mm Cap Screw Type B 120 mm Cap Screw Type C Cap Liner D Shaft Retaining Nut BP 3401 xxx Impeller with Glass Shaft Magnetic Complete E Paddle Bearing Holder F Paddle Blade G Paddle Polysulfone Rod 125 ml 1L Flasks H Paddle Glass Rod 3L 36L Flasks I Paddle s s Rod Optional 125 mL 36L Flasks J Paddle Retaining Sleeve K Paddle Magnet L Paddle Silicone Plug M PTFE Split Ring 1 per Glass Rod 2 per Polysulfone or s s Rod C NN SO A em DIMMI 1 Em EA Fe x p oe fe M r K Figure 33 ProCulture 4500 amp 4502 Series Flask Pa
59. s Flask with 100 120mm Center Neck Opening Vertical Sidearms Part Description 4510 12 xxx Flask with 100 120 mm Center Neck Opening Vertical Sidearms A Replacement Flask with 100 120 mm Center Neck Opening 402614 400649 100 120 mm Cap Assembly Complete Direct Drive Style Shaft O ring Fitting Male Luer x 1 4 28 Polypropylene 13mm 0 2 micron PTFE Filter Snap Ring Not Shown B 100 120 mm Cap Screw Type C Cap Lock Nut D Cap Liner E Drive Pin F Spacer G Bearing Upper Iorlon H Polymer Bearing Lower 1 4 x 5 8 I Seal Assembly Direct Drive J Nut Luer Lock Polypropylene K Shaft Housing L S S Shaft M N O D B z E SA 7 TaN SDS ra o ora ora NA A i LIRE SSR N AT SOO ieS tT a ER pama LOOOTO y y IL LOOO TOOT C ill L di Bi EL Stainless Steel Coupling Collar Corning OL USA 8000 Gap Paddle bik A Assembly Figure 35 ProCulture Direct Drive Detail with 100 120mm Cap Assembly PROCULTURE SPINNER FLASKS OPERATING INSTRUCTIONS 33 Flask Details ProCulture Spinner Flasks 4510 12 series 100 120mm Center Neck Flask Fitted with 402614 400649 Direct Drive See Figure 36 4510 12 Series Flask with 100 120 mm Center Neck Opening Vertical Sidearms Part Description 4510 12 xxx Flask with 100 120 mm Center Neck Opening Vertical Sidearms A Replacement Flask with 100 120mm Center Neck Opening
60. s leave a residue which can dissolve into cell culture media and make them alkaline The use of a laboratory scrub brush is acceptable but take care to check it for wear to prevent scratching the glass A plastic bristle brush is recom mended Thoroughly rinse the vessel and all components immediately with distilled or deionized water Avoid letting the detergent dry on the surface of any parts Allow parts to dry before assembling When thoroughly dry carefully assemble the vessel according to the case insert diagram Siliconizing Glassware The basement membrane produced by many attachment dependent cell lines grown on microcarrier beads may adhere to the glassware where it can complicate subsequent cell culture In addition proteinaceous material from cells or media may accumulate at the gas liquid interface creating a crust that can encourage further deposits It is best to try and minimize its buildup on the wall of the vessel by placing a layer of silicone on the glassware Several products on the market are available to accomplish this such as SigmaCote Sigma Chemical Company Prosil 28 PCR Research or Repelcote Hopkins amp Williams It is best to follow the rec ommendations of the manufacturer as to their use Be sure to thoroughly wash the vessel after treatment prior to its use in cell culture One treat ment is usually sufficient for many cell culture cycles Another treatment is necessary when a noticeable bu
61. ss Shaft Magnetic Complete 36000 mL Impeller Assembly Components Bearing Holders BP 3411 010 BP 3411 011 BP 3411 012 BP 3411 013 BP 3411 014 BP 3411 015 BP 3411 016 BP 3411 017 BP 3411 018 Paddle Blades BP 3411 020 BP 3411 021 BP 3411 022 BP 3411 023 BP 3411 024 BP 3411 025 BP 3411 026 BP 3411 027 BP 3411 028 BP 3411 9022 BP 3411 9023 BP 3411 9024 BP 3411 9025 BP 3411 9026 BP 3411 9027 BP 3411 9028 Paddle Bearing Holders 125 mL Paddle Bearing Holders 250 mL Paddle Bearing Holders 500 mL Paddle Bearing Holders 1000 mL Paddle Bearing Holders 3000 mL Paddle Bearing Holders 6000 mL Paddle Bearing Holders 8000 mL Paddle Bearing Holders 15000 mL Paddle Bearing Holders 36000 mL Paddle Blades 125 mL Paddle Blades 250 mL Paddle Blades 500 mL Paddle Blades 1000 mL Paddle Blades 3000 mL Paddle Blades 6000 mL Paddle Blades 8000 mL Paddle Blades 15000 mL Paddle Blades 36000 mL Paddle Blades 500 mL Mod Probes Paddle Blades 1000 mL Mod Probes Paddle Blades 3000 mL Mod Probes Paddle Blades 6000 mL Mod Probes Paddle Blades 8000 mL Mod Probes Paddle Blades 15000 mL Mod Probes Paddle Blades 36000 mL Mod Probes Paddle PTFE Split Ring Thrust Bearing Washers BP 3411 030 BP 3411 031 Paddle Split Ring Thrust Bearing Washers for 1 4 O D Stainless Steel Shaft Paddle Split Ring Thrust Bearing Washers for 5 16 O D Stainless Steel Shaft For ordering and technical service of all BP 3xxx xxx part num
62. tional Information Application Check List l 22 Select the Unit Operation s necessary from the Schematic Circuit illustrations on pages 28 and 29 and the list below Continuous Sparging Controlled Sparging pH Monitoring Control Temperature Monitoring Control O Monitoring Control Media Inoculum Handling In line Microporous or Ultrafiltration Vessel Venting Find the desired Unit Operation Schematic Circuit Diagram from the illustrations which most closely matches your needs and jot down the fittings for that operation Specific listings of components are summa rized in the brochure Determine the number of sidearms on the vessel necessary to accom plish the Unit Operations desired Typically each fitting requires one free sidearm port Keep in mind that if the number of sidearm fittings needed exceeds the number of sidearms on the vessel selected it is often possible to rearrange the fittings on a sidearm port to provide more than one Unit Operation on that sidearm Select the type of probe and penetration style if any There are two basic styles Threaded probe with without mounting fitting Insertion probe Note Contact manufacturer for information on the type and size of the mounting fixture Obtain prints if possible Most Ingold Mettler Toledo or Broadley James products can be supported by Corning Contact Corning Technical Support Department 1 800 492 1110 in the U S Outside

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