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ROTORS AND TUBES

1. Figure 5 1 Examples of Swinging Bucket Rotors 5 2 J Series Rotors and Tubes Using Swinging Bucket Rotors Table 5 1 General Specifications for Beckman Coulter J Series Swinging Bucket Rotors JS 24 38 24 000 rpm N A 161 0 118 2 75 3 6 x 38 5 mL 231 mL 110 500 x g Ht 334 JS 24 15 24 000 rpm N A 171 3 1221 72 9 6x15 mL 90 mL 103 900 x g Al 376 JS 13 1 13 000 rom 400 140 91 41 6 x 50 mL 300 mL 26 500 x g to 1841 1450 JS 7 5 7500 rpm 600 165 108 51 4x 250 mL 1 liter 10 400 x g to 1090 800 JS 5 9 5900 rpm 500 194 8 179 6 164 3 10 microplates 384 mL 6570 x g to 4 deep well plates 1200 2 squarewell plates JS 5 3 5300 rom 500 168 5 153 4 138 6 24 microplates 768 mL 6130 x g to 8 deep well plates deep well plates 1200 4 square well plates 6870 x g 218 4 155 6 92 7 4 x 500 mL 2 liters 500 mL bottles JS 5 2 5200 rpm 600 226 156 86 4 x 1 liter 4 liters 6840 x g to 4 blood bags 9051 800 12 microplates 148 RIA tubes JS 5 0 5000 rpm 300 267 188 108 4 x 2 25 liters 9 liters 7480 x g to 9171 600 Maximum speeds are based on a solution density of 1 2 g mL t Critical speed range is the range of speeds over which the rotor shifts so as to rotate about its center of mass Passing through or running at the critical speed range is characterized by some vibration J Series Rotors
2. Maximum speeds are based on a solution density of 1 7 g mL for the JV 20 rotor and 1 2 g mL for the JR 3 2 rotor Critical speed range is the range of speeds over which the rotor shifts so as to rotate about its center of mass Passing through or running at the critical speed range is characterized by some vibration min lav max Axis of Rotation 4 Figure 6 1 Vertical Tube Rotor RACK TYPE ROTORS The rack type rotor see Figure 6 2 holds a wide range of gamma counter tubes in tube racks Racks are loaded into removable trays which are then loaded into carriers at a resting angle During centrifu gation the carriers swing out to a completely horizontal position to provide uniform pelleting of samples The nearly vertical position of the racks during centrifugation permits processing of up to 320 mL in one run 6 2 J Series Rotors and Tubes Imin lav Tmax Axis of Rotation Using Vertical Tube and Rack Type Rotors Figure 6 2 Rack Type Rotor I gt NOTE Although rotor components and accessories made by other manufacturers may fit in the Beckman Coulter rotor you are using their safety in the rotor cannot be ascertained by Beckman Coulter Use of other manufacturers components or accessories in the Beckman Coulter rotor may void the rotor warranty and should be prohibited by your laboratory safety
3. essen 2 4 Polycarbonate Tubes 0 0 eee ee eee 2 5 Polypropylene Tubes 0 0 cee eee eee eee ee 2 5 Polyethylene Tubes 0 0 cece eee eee 2 5 Ultra Clear Tubes 5 2 e ec paves Rem ems 2 6 Stainless Steel Tubes 0 0 0 0 eee eee eee eee 2 6 Microfuge TUBES 21 be bee ea a n 2 6 Bottles cepere ER sabe teed INE ewe e 2 7 Multiwell Titer Plates 2 7 Temperature Limits 0 0 0 eee eee e 2 8 iii Contents iv SECTION 3 Page Spacers and Floating Spacers 0 0 0 cece eee eee 2 9 Adapters ihc iee Perdew hehe EC ome E RL RR deen 2 9 Bottle Adapters racerene aa sei ERO RR ERR 2 9 Multitube Adapters 0 0 0 cee eee eee 2 10 Bottle and Tube Caps sees 2 11 Aerosolve Cannisters 0 2 eee eee eee 2 12 Blood Bag Cups 2 0 cece eee e 2 12 Rotor Labware Assemblies 00 000 e eee ee eee 2 13 USING TUBES BOTTLES AND ACCESSORIES Gradient Preparation 0 eee eee eee 3 1 General Filling and Sealing or Capping Requirements 3 2 Working with Physiological Fluids LL 3 4 Filling Open Top Tubes eese 3 5 Open Top Polyallomer Tubes 0 000005 3 5 Other Open Top Tubes 0 0 0 0 ee eee eee 3 5 Capping TUDES a ue ea ee epo EE oT aa tinge ate 3 7 Filling and Capping Bottles 2 0 0 0 00 eee eee eee ee 3 7 Three Piece Cap Assemblies 00
4. MULTIWELL TITER PLATES Titer plates can be run in specially designed carriers in some swinging bucket rotors Carriers are used by installing them on the pivot pins in place of the buckets normally used with the rotor or in buckets designed to run plates Because the plates can break under the stresses of high speed centrifugation speed reduction is usually required when running multiwell plates Multiwell plates are also used in adapters in the rack type rotor 96 Well Microtiter Plates The 96 well plates are manufactured of specially formulated poly styrene These flat bottom nonsterile plates normally hold 300 uL per well of sample and solvent Volume J Series Rotors and Tubes 22 Tubes Bottles and Accessories Deep Well Titer Plates and Caps ae 96 Cap Strip These plates are manufactured of sterile or nonsterile polystyrene Z CEECC EOM gt ue or polypropylene The plates can contain up to 1 2 mL per well of QE sample and solvent in a single 96 well plate when run uncapped Ix CERCA S E CEU eto When used with caps which come in 96 cap strips each well Q Wi 5 ro 0 0 accommodates 1 0 mL Square Well Titer Plates Square well plates are manufactured of nonsterile polypropylene The square well format provides 2 mL per well capacity in each 96 well plate Working Volume TEMPERATURE LIMITS Each labware material has a specified temperature range Although som
5. Plug 343685 Carrier Sealing Surface Aluminum Composite Interface _ R mme Outside Wall Sleeve Washer LUBRICATION O Rings J Series Rotors and Tubes Care and Maintenance Regularly check the condition of O rings or gaskets and replace any that are worn or damaged Regularly check that all sealing surfaces are smooth and undamaged to ensure proper sealing Before each use inspect Aeroseal cover sealing surfaces especially the O ring groove It must be smooth and free of scratches Also ensure that the top 2 54 cm 1 in of the bucket is clean and smooth buckets with scratches or gouges in this surface will not seal properly Inspect the O ring and plug for nicks abrasions and other damage Replace damaged components with Beckman Coulter parts only do not use a substitute for the O ring it has been specifically selected for this application Regularly check the condition of the Micro Plus carrier base and pad and do not use them if there are signs of damage Retire the base from use after 1 year Before each use inspect carbon fiber cannisters for cracks where carbon fiber threads are visible If any cracking or other damage is visible on the outside wall or near the aluminum composite inter face area do not use the cannister Contact your Beckman Coulter representative Retire the cannister on the expiration date Regular and thorough lubrication can extend the useful lif
6. ADSOL angular velocity anodized coating autoclaving buoyant density Buna N centrifugal effect centrifugal force J Series Rotors and Tubes Appendix E Glossary of Terms dextrose sodium chloride mannitoladenine an additive used as a stabilizer for red blood cells that extends red cell life in CPD rate of rotation measured in radians per second 2nrpm o or 0 10472 rpm a thin hard layer of aluminum oxide formed electrochemically on aluminum rotor and or accessory surfaces as a protective coating for corrosion resistance sterilization by heat dry or steam the density of a particle in a specified liquid medium black nitrile rubber used for O rings and gaskets in rotor assemblies should be used at temperatures between 34 and 121 C 30 and 250 F accumulated value of t2 a dt t1 where f is time and is angular velocity in a centrifugal field the force which causes a particle to move away from the center of rotation E 1 Glossary of Terms E 2 clearing factor k clearing time t CPD CPDA 1 cryoprecipitate CsCl CsSO Delrin density density separation differential separation EPDM erythrocytes calculated for all Beckman Coulter high speed rotors as a measure of the rotor s relative pelleting efficiency In tinay Zr k max min 10 a 3600 or min RPM 1000 253303 x In rg n t k s where
7. Authorized dealers in other countries 4300 N Harbor Boulevard Box 3100 Fullerton California 92834 3100 Sales 1 800 742 2345 Service 1 800 551 1150 Internet www beckmancoulter com Telex 678413 Fax 1 800 643 4366 CA ce Printed on recycled paper 2002 Beckman Coulter Inc e LS EN ISO 9001
8. Maximum speed in an Avanti J E for the rotor with magnets without magnets maximum is 14 000 rpm Maximum speed at 2 C in a 50 Hz centrifuge is 14 000 rpm 4 Maximum speed in an Avanti J E for the rotor with magnets without magnets maximum is 13 000 rpm Maximum speed at 2 C in a 50 Hz centrifuge is 15 000 rpm Maximum speed in an Avanti J series centrifuge Maximum speed in a J2 series centrifuge is 14 000 rpm Maximum speed in an Avanti J E for rotor without magnets is 6 300 rpm Maximum speed for rotor in an Avanti J E centrifuge is 6 300 rpm h The JS 24 38 and JS 24 15 rotors can achieve 24 000 rpm in an Avanti J 30 I centrifuge only In Avanti J 25 series and J 20 series centrifuges the maximum speed for these rotors is 10 000 rpm J Series Rotors and Tubes 1 5 Rotors 1 6 Table 1 1 Rotors Used in Beckman Coulter J Series Centrifuges continued Rotors in parentheses are no longer manufactured Fixed Angle continued JA 18 1 liter 18 000 47 900 566 X X X X X X X X X X JA 17 700 mL 17 0004 39 800 690 X X X X X X X X X X JLA 16 250 1 5 liter 16 000 38 400 1090 X X X X X X JA 14 1 5 liter 14 000 30 100 1764 X X X X X X X X X X JA 12 600 mL 12 000 23 200 1244 X X X X X X X JA 10 3 liters 10 000f 17 700 3610 X X X X X X X X X X X JLA 10 500 3 liters 10 000 18 500 2850 X X X X X X X JLA 9 1000 4 liters 9 0002 16 800 2540 X X X X JLA 8 1000 6 liters 8 000 15 9
9. Sodium Hypochlo Hydrogen Glutaral Tube Bottle Autoclave UV Ethylene Formal Ethanol rite Peroxide dehyde Phenolic Material 121 C Irradiation Oxide dehyde 70 10 10 2 Derivatives polyallomer yes no yes yes yes yes yes yes no Ultra Clear no no yes yes yes yes yes yes no polycarbonate yes no yes yes no yes yes yes no polypropylene yes no yes yes yes yes yes yes no polyethylene no no yes yes yes yes yes yes yes cellulose propionate no no no no no yes yes yes no stainless steel yes yes yes yes yes no yes yes no To avoid deformation autoclave tubes or bottles upside down in a tube rack at 15 psig for no more than 20 minutes allow to cool before removing from rube rack DO NOT autoclave capped or sealed tubes or bottles Flammable do not use in or near operating centrifuges 3 Do not use if there is methanol in the formula Tube life will be reduced by autoclaving 5 Discoloration may occur 6 Can be used if diluted 7 Below 26 C only 8 Below 21 C only Marginal 7 8 J Series Rotors and Tubes 121 C Eo nr J Series Rotors and Tubes Care and Maintenance Most tubes and accessories except those made of Ultra Clear poly ethylene or cellulose propionate can be autoclaved at 121 C for about 30 minutes Note that autoclaving reduces the lifetime of polycarbonate tubes Also polyallomer tubes may be permanently deformed if they are a
10. Subject to the conditions specified below and the warranty clause of the Beckman Coulter Inc terms and conditions of sale in effect at the time of sale Beckman Coulter Inc agrees to correct either by repair or at its election by replacement any defects of material or workmanship which develop within seven 7 years after delivery of a J series rotor to the original buyer by Beckman Coulter Inc or by an authorized represen tative provided that investigation and factory inspection by Beckman Coulter Inc discloses that such defect developed under normal and proper use Should a Beckman Coulter cen trifuge be damaged due to a failure of a rotor covered by this warranty Beckman Coulter will supply free of charge all cen trifuge parts required for repair REPLACEMENT Any product claimed to be defective must if requested by Beckman Coulter Inc be returned to the factory transporta tion charges prepaid and will be returned to Buyer with the transportation charges collect unless the product is found to be defective in which case Beckman Coulter Inc will pay all transportation charges A defective rotor will be replaced by Beckman Coulter Inc at its then current list price less a credit based upon the age of the rotor years since date of purchase The Buyer shall not receive credit until the claimed defective rotor is returned to Beckman Coulter s Palo Alto California facility or delivered to a Beckman Field Service r
11. 1 Select the appropriate adapter base and attach a bail to it 2 Place the base and bail in an empty bucket or on the lab bench not in the rotor 3 Position one of the disks so that its grooves are aligned with the bail Push the disk down until the bail snaps into the grooves 4 Add more disks until the height of the assembly is nearly as tall as the tubes you will be using If the height of the disks is very tall you may have to push the bail into the grooves of the top disks by hand Remove or add disks to the bail to accommodate shorter or longer tubes If the tubes fit too snugly in the adapter s rubber base apply a light film of dusting power such as talcum powder to prevent the tubes from sticking Place each tube in an adapter so that its weight is balanced by a tube in a diametrically opposite position across the pivotal axis in the same adapter Adapters placed in opposing buckets should also be filled the same way see Figure 5 2 If you must run only one tube in an adapter be sure this tube rests over the bucket s pivotal axis gt NOTE Be sure to run a tube of the same approximate weight in the same configuration in the opposite bucket J Series Rotors and Tubes 5 11 Using Swinging Bucket Rotors Using Blood Bag Cups Do not pour liquid directly into blood bag cups Fit blood bags into cups before loading the cups into the rotor buckets Load the blood bag cups as follows 1 Load the cups so tha
12. Density Refractive by mg mL of Density Refractive by mg mL of g cm3 Index nD Weight Solution Molarity g cm3 Index nD Weight Solution Molarity 1 0047 1 3333 1 10 0 0 056 1 336 1 3657 34 454 2 2 698 1 0125 1 3340 2 20 2 0 119 1 3496 1 3670 35 472 4 2 806 1 0204 1 3348 3 30 6 0 182 1 363 1 3683 36 490 7 2 914 1 0284 1 3356 4 41 1 0 244 IR 1 3696 37 509 5 3 026 1 0365 1 3364 5 51 8 0 308 1 391 1 3709 38 528 6 3 140 1 0447 1 3372 6 62 8 0 373 1 406 1 3722 39 548 3 3 257 1 0531 1 3380 7 73 7 0 438 1 4196 1 3735 40 567 8 3 372 1 0615 1 3388 8 84 9 0 504 1 435 1 3750 41 588 4 3 495 1 0700 1 3397 9 96 3 0 572 1 450 1 3764 42 609 0 3 617 1 0788 1 3405 10 107 9 0 641 1 465 1 3778 43 630 0 3 742 1 0877 1 3414 11 119 6 0 710 1 481 1 3792 44 651 6 3 870 1 0967 1 3423 12 131 6 0 782 1 4969 1 3807 45 673 6 4 001 1 1059 1 3432 13 143 8 0 854 1 513 1 3822 46 696 0 4 134 1 1151 1 3441 14 156 1 0 927 1 529 1 3837 47 718 6 4 268 1 1245 1 3450 15 168 7 1 002 1 546 1 3852 48 742 1 4 408 1 1340 1 3459 16 181 4 1 077 1 564 1 3868 49 766 4 4 552 1 1437 1 3468 17 194 4 1 155 1 5825 1 3885 50 791 3 4 700 1 1536 1 3478 18 207 6 1 233 1 601 1 3903 51 816 5 4 849 1 1637 1 3488 19 221 1 1 313 1 619 1 3920 52 841 9 5 000 1 1739 1 3498 20 234 8 1 395 1 638 1 3937 53 868 1 5 156 1 1843 1 3508 21 248 7 1 477 1 658 1 3955 54 859 3 5 317 1 1948 1 3518 22 262 9 1 561 1 6778 1 3973 55 922 8 5 481 1 2055 1 3529 23 277 3 1 647 1 699 1 3992 56 951 4 5 65
13. J Series Rotors and Tubes Sandwich Coiled Tubing between Transfer Bags and Filled Blood Bag Blood Bag Cup SAMPLE RECOVERY J Series Rotors and Tubes Using Tubes Bottles and Accessories Different cups color coded for capacity identification can accom modate single double triple or quad pack blood bags Refer to the applicable rotor manual to determine the correct blood bag cup to use Do not pour liquid directly into blood bag cups Fit blood bags into cups before loading the cups into the rotor buckets Stuffing blood bags directly into the rotor while it is installed in the centrifuge can trip the imbalance detector 1 Load the bags as far down into the cups as possible Make sure the bags stay as vertical as possible with no folds at the top or corners If folds are present blood cells could remain in the folds and then mix with the plasma when the bag is removed 2 Sandwich the tubing between the blood bag and any transfer packs 3 Make sure the loaded blood bag cups opposite each other on the rotor yoke are approximately the same weight within 1 gram Balancing pads can be used with some rotors if necessary to maintain weight balance Load blood bag cups into the rotor buckets To reduce the possibility of bag breakage align the blood bag seam with the rotor pivot pins with the label facing out away from the axis of rotation N CAUTION If disassembly reveals evidence of leakage you should
14. JS 5 2 JS 4 2 JS 4 2A JS 4 0 and JS 3 0 Rotors Lubricate the O ring and plug of Aeroseal bucket covers with silicone vacuum grease Also lightly grease the inside top 1 27 cm 0 5 in of the bucket FIELD ROTOR INSPECTION PROGRAM The Field Rotor Inspection Program FRIP has two purposes to prevent premature rotor failures by detecting conditions such as stress corrosion metal fatigue damage or wear in the anodized coatings and to instruct laboratory personnel in the proper care of rotors J Series Rotors and Tubes Care and Maintenance Beckman Coulter has trained a group of experienced service engineers in the techniques of nondestructive evaluation For more information about the program contact your Beckman Coulter representative TUBE BOTTLE AND ACCESSORY CARE Proper care of tubes and bottles involves observing temperature fill volume and run speed limitations as well as careful cleaning and sterilization procedures CLEANING Do not wash tubes and bottles in a commercial dishwasher detergents and temperatures are too harsh Wash tubes bottles adapters and blood bag cups by hand using a mild detergent such as Solution 555 339555 diluted 10 to 1 with water and a soft brush Disassemble multitube adapters for cleaning After washing with Solution 555 and a soft brush rinse them with water then dry and reassemble e Polycarbonate bottles and tubes are vulnerable to attack by
15. ROTOR LABWARE ASSEMBLIES AirVent amp Filter N 1 a RN ge Plug Cup Cover HarvestLine System Liner Green Gasket 369261 J Series Rotors and Tubes The JS 5 0 labware assembly has an available HarvestLine system liner If liners are not used the sample can be loaded directly into the cup and a partition can be inserted to minimize sample disturbance at low g forces The gasket and the cup and cover surfaces that contact the gasket must be dry to ensure sealing Gasket 36926 green is used when liners are used gasket 369257 red is used when the cup is used alone with or without a partition The cup cover top surface can be written on to identify the assembly or sample The HarvestLine System for the JLA 8 1000 and JLA 9 1000 rotors provides a convenient method of loading recovering and storing samples run in these rotors Up to six rotor bottles are placed in the filling rack and a liner is placed into each bottle The liners are loaded with sample through a funnel or fermentor hose The valve in the neck of each liner is then sealed and the liner necks folded to fit inside the bottles The bottles are sealed with rotor plugs and cap closures and the sealed bottles are placed into the rotor cannisters for centrifugation After centrifugation the liner valves are cut off and the supernatant decanted either for storage or disposal The liners can t
16. Two factors affect symmetric loading e The buckets or carriers must be loaded symmetrically with respect to their pivotal axes the pivotal axis runs parallel to the crossbar see Figure 5 2 The rotor should be loaded symmetrically with respect to its center of rotation J Series Rotors and Tubes Using Swinging Bucket Rotors Balanced Load Unbalanced Load Pivotal Axis of Bucket Center of Rotation Correct Incorrect eeooooee 00000000 Oooeeeeco OOOOOOOO eoooo O eeooooee 00000000 Oooeeeeco OOOOOOOO eoooo O eeooooee OOOOOOOO Oooeeeeoco OOOOOOOO eoooo O eeooooee OOOOOOOO OoOeeeeoco OOOOOOOCO eoooo O eeooooee OOOOOOOO Oooeeeeco o e eoooo O eeooooee OOOOOOOO Oooeeeeco e e eoooo o eeooooee OOOOOOOO Oooeeeeco e e eoooo e eeooooee OOOOOOOO Oooeeeeoco e e eoooo e eeooooee OOOOOOOO Oooeeeeco OOOOOOOO eoooo e eeooooee OOOOOOOO Ooeeeeco OOOOOOOO eoooo e eeooooee 00000000 Oooeeeeco OOOOOOOO eoooo e eeocoooee eoccc0000 Oooeeeeoco OOOOOOOO eoooo Examples of Symmetrically Loaded Trays Examples of Nonsymmetrically Load Opposite Trays the Same Way Loaded Trays Pivotal Axis N Carrier Ooooooooooooo OOOOOOOOOOOO 000000000008 Figure 5 2 Examples of Correctly and Incorrectly Loaded Buckets and Carriers Contents of opposing buckets must be the same and each bucket must be balanced on its pivotal axis J Series Rotors and Tubes 5 9 Using Swinging Bucket
17. officer Only the components and accessories listed in the applicable rotor manual should be used USING A VERTICAL TUBE ROTOR TUBES AND BOTTLES J Series Rotors and Tubes Only Quick Seal tubes listed in the rotor manual may be centrifuged in a vertical tube rotor Refer to Section 3 USING TUBES AND BOTTLES for filling and sealing requirements of Quick Seal tubes Observe the maximum rotor speeds and fill volumes listed in the rotor manual Using Vertical Tube and Rack Type Rotors ROTOR PREPARATION AND LOADING For runs at other than room temperature refrigerate or warm the rotor beforehand for fast equilibration Prerun Safety Checks Read all safety information in the rotor manual before using the rotor 1 Make sure that the rotor and plugs are clean and show no signs of corrosion or cracking 2 Check the chemical compatibilities of all materials used Refer to Appendix A CHEMICAL RESISTANCES 3 Verify that the tubes spacers and floating spacers being used are in good condition and are listed in the rotor manual Rotor Preparation 1 Be sure that the plug threads are lightly but evenly lubricated with Spinkote lubricant 306812 2 Lubricate the rotor drive hole with silicone vacuum grease 335148 3 Load the filled and sealed tubes symmetrically into the rotor Opposing tubes must be filled to the same level with liquid of the same density Refer to ROTOR BALANCE in Section 1 emis Spacer 4
18. 1 S 10 5 seconds swinging bucket rotor a rotor in which the tubes or bottles are carried in buckets microtiter plate carriers or racks that swing up to the horizontal position during centrifuga tion sometimes referred to as a horizontal or swing out rotor Ultem polyetherimide PET used in adapters covers and spacers should be used at temperatures between 29 and 204 C 20 and 400 C Ultem is a regis tered trademark of GE Plastics vertical tube rotor a rotor in which the tubes or bottles are held parallel to the axis of rotation Viton fluorocarbon elastomer used in high temperature applications Viton is a registered trademark of E I Du Pont de Nemours amp Company WBC white blood cells or leukocytes protect the body against infection and many diseases wettable tube or bottle material that water or other aqueous solution will adhere to the more wettable a tube or bottle material is the more biological material DNA protein cells and so forth will adhere to the walls E 6 J Series Rotors and Tubes IN 175 IN 181 IN 192 A 1792 BR 8102 DS 528 DS 602 DS 605 DS 719 J Series Rotors and Tubes Appendix F References Documents referenced below are available upon request from Beckman Coulter Inc Technical Publications Department 1050 Page Mill Road Palo Alto CA 94304 U S A Telephone 650 859 1753 Fax 650 859 1375 Chemical Resistances How to use Quick Seal Tu
19. 600 to 32 x 15 mL 480 mL 23 Angle 51500xg 800 115 107 64 outer row 43 900 x g 98 73 47 inner row 371 Continued a Maximum speeds are based on a solution density of 1 2 g mL in all rotors except for the JA 18 1 which is rated for a density of 1 4 g mL b Critical speed range is the range of speeds over which the rotor shifts so as to rotate about its center of mass Passing through or running at the critical speed range is characterized by some vibration Maximum speed in an Avanti J E centrifuge is 21 000 rpm 4 When a JA 18 1 rotor is used in the J2 HC centrifuge derate the rotor as follows when the 45 adapters are used do not run the rotor above 15 000 rpm when 25 adapters are used do not run the rotor above 16 000 rpm Maximum speed in an Avanti J series centrifuge except Avanti J E maximum speed in an Avanti J E for a rotor with magnets maximum speed for rotor without magnets is 13 000 rpm Maximum speed in a J2 series centrifuge is 14 000 rpm Maximum speed in an Avanti J E for a rotor with magnets maximum speed for a rotor without magnets is 13 000 rpm g Maximum speed in an Avanti J E for the rotor with magnets without magnets maximum is 14 000 rpm Maximum speed at 2 C in a 50 Hz centrifuge is 14 000 rpm Do not put bottles directly into the rotor without cannisters J Series Rotors and Tubes 4 3 Using Fixed Angle Rotors Table 4 1 General Specifications for Beckman Coulter J Series F
20. 7 2 401 1 1270 1 3811 30 338 1 0 988 1 3103 1 4509 64 838 6 2 450 1 1318 1 3829 31 350 9 1 025 1 3163 1 4532 65 855 6 2 500 1 1366 1 3847 32 363 7 1 063 1 3224 1 4558 66 872 8 2 550 1 1415 1 3865 33 376 7 1 100 1 3286 1 4581 67 890 2 2 864 Divide by 10 0 to obtain 96 w v Density and refractive index data are from the International Critical Tables C 4 J Series Rotors and Tubes Table C 4 Density Conversion for Cesium and Rubidium Salts at 20 C Gradient Materials 26 w w CsCl CsBr Csl Cs2S04 CsNO3 RbCI RbBr Rbl Rb2SO4 RbNOs 1 1 00593 1 00612 1 00608 1 0061 1 00566 1 00561 1 00593 1 00591 1 0066 1 0053 2 1 01374 1 01412 1 01402 1 0144 1 01319 1 01307 1 01372 1 01370 1 0150 1 0125 i 1 02969 1 03048 1 03029 1 0816 1 02859 1 02825 1 02965 1 02963 1 0322 1 0272 6 1 04609 1 04734 1 04707 1 0494 1 04443 1 04379 1 04604 1 04604 1 0499 1 0422 8 1 06297 1 06472 1 06438 1 0676 1 06072 1 05917 1 06291 1 06296 1 0680 1 0575 10 1 08036 1 08265 1 08225 1 0870 1 07745 1 07604 1 08028 1 08041 1 0864 1 0731 12 1 09828 1 10116 1 10071 1 1071 1 09463 1 09281 1 09817 1 09842 1 1052 1 0892 14 1 11676 1 12029 1 11979 1 1275 1 11227 1 11004 1 11661 1 11701 1 1246 1 1057 16 1 13582 1 14007 1 13953 1 1484 1 12775 1 13563 1 13621 1 1446 1 1227 18 1 155
21. 8 E 6 5 3 SA 15 000 6 6 B 4 4 3 N 10 000 3 2 E 4 4 0 N 20 000 N N 10 9 9 9 7 18 000 N N E E 6 4 xol 15 000 7 5 4 4 4 3 N 10 000 2 E 4 0 0 0 N 20 000 N N 9 8 7 E 6 18 000 N N 6 6 5 4 8 JA20 15 000 2B 4 4 4 3 2 E 10 000 2 2 4 0 0 0 0 18 000 N N N N 8 5 0 17 000 N N N N 6 5 0 JA 18 1 15 000 N N 9 B 4 3 0 10 000 7 7 3 8 2 0 42 8 000 6 B 3 3 4 0 42 48 000 N N N N N 10 7 17 000 N N N 10 9 8 6 16 000 N N 10 10 9 E 5 rs 12 000 N 6 6 6 5 4 3 8 000 E 4 4 3 2 4 0 5 000 4 3 2 2 4 0 0 17 000 N N E m E 4 2 15 000 N 5 4 4 4 3 4 aid 12 000 4 4 3 3 3 E N 8 000 4 4 0 0 0 0 N 14 000 N N 7 E 4 3 3 JLA 16 250 10000 N 4 3 8 2 E N 5 000 N 0 0 0 0 0 N Above 30 C ambient temperature this temperature may not be achieved at this speed Continued N indicates that the rotor cannot achieve the desired temperature at this speed J Series Rotors and Tubes Temperature Compensation Tables Table B 2 Temperature Compensation Settings for the J2 21 J2 21B J2 21C and J2 HS Centrifuges continued 14 000 N N 7 7 7 5 4 12 000 N 6 5 5 4 3 2 ats 10 000 4 4 3 3 2 E N 5 000 1 0 0 0 0 0 N 12 000 N 6 6 7 7 6 9 JA 12 10 000 3 4 5 5 5 5 9 5 000 1 3 3 4 4 5 9 10 000 N 3 2 1 0 1 2 JA 10 8 000 3 2 1 1 0 1 2 5 000 2 0 0 0 0 1 2 10 000 N 5 2 1 0 2 1 J
22. J Series Rotors and Tubes J Series Rotors and Tubes Figure 1 1 Figure 1 2 Figure 1 3 Figure 1 4 Figure 1 5 Figure 1 6 Figure 3 1 Figure 4 1 Figure 5 1 Figure 5 2 Figure 5 3 Figure 5 4 Figure 6 1 Figure 6 2 Figure D 1 Illustrations Page Fixed Angle Swinging Bucket Vertical Tube and Rack Type Rotors leeleeeeeeee n 1 2 Particle Separation in Fixed Angle Swinging Bucket and Vertical Tube Rotors 2 esee erae rat erbe et 1 8 Sedimentation Coefficients in Svedberg Units for Some Common Biological Materials llle 1 10 Nomogram for J2 Series Centrifuges llle eese 1 11 Nomogram for J6 Series Centrifuges 00000000 1 12 Arranging Tubes Symmetrically in a Fixed Angle Vertical Tube or JS 24 Series Swinging Bucket Rotor 1 15 The Cordless Quick Seal Tube Topper 000000 3 10 Examples of Fixed Angle Rotors 0 0 00 0s eee 4 2 Examples of Swinging Bucket Rotors 000 5 5 2 Examples of Correctly and Incorrectly Loaded Buckets and CAMELS sy ss ees PEU LE a aud ad tba Sed Gea 5 9 Typical Blood Bag Loading Procedure JS 4 3 Rotor Shown 5 12 The Micro Plus Microtiter Plate Carrier Base Pad and Deep Well Microtiter Plate 5 15 Vertical Tube Rotor oee sorei st ee eee E A 6 2 Rack Typ Rotor 2 oe cca dene per RR ead aes 6 3 Blood Component Preparation 0 0 0 cece eee eee D 5 V
23. Rotors At Rest At Speed rej ra Proper Weight Distribution r ra rors Improper Weight Distribution 5 10 This means that for best results you should load opposing buckets or carriers with the same type of labware containing the same amounts of fluid of equal density Additionally buckets or carriers placed opposite each other on the rotor yoke must balance to within a certain weight typically 10 grams see the applicable rotor manual for details Do not exceed the rated maximum load for buckets or carriers IB NOTE The JS 4 2A and JS 4 2SMA swinging bucket rotors incorporate ARIES Automated Rotor Imbalance Equilibrating System Smart Balance technology which provides imbalance compensation for rotors with buckets that are up to 100 grams unbalanced due to different loading volumes or tube or bag breakage Beckman Coulter supplies buckets and carriers for most rotors in weight matched sets to make balancing easier the weight and date of manufacture are marked on the side of each bucket and bottom of each carrier Modular disk adapters are also sold in weight matched sets However there are variances in weight between sets as well as variance in weight between previously purchased adapters To prevent accidental imbalance it is important to keep matched sets of adapters together and to weigh other adapters to be sure they are approximately the same Marking matche
24. alkaline solutions and detergents so use a detergent with pH less than 9 such as Solution 555 Do not use a brush with exposed metal scratches in polycarbonate will cause early failure Alcohol and acetone react unsatisfactorily with many tube and accessory materials If a solvent must be used to rinse dry or decontaminate these materials consult Appendix A to select an appropriate solvent Do not dry tubes bottles or accessories in an oven Labware should be air dried e Quick Seal Ultra Clear and thinwall polyallomer tubes are intended for one time use and should be discarded after use J Series Rotors and Tubes 9 7 Care and Maintenance DECONTAMINATION C Labware contaminated with radioactive or pathogenic solutions CAN should be decontaminated or disposed of following appropriate safety ae guidelines and or regulations Consult Appendix A to select an agent that will not damage the tube or bottle material STERILIZATION AND DISINFECTION Refer to Table 7 1 for sterilization methods recommended for each container type Table 7 1 Tube and Bottle Sterilization and Disinfection This information is provided as a guide to the use of sterilization and disinfection techniques for tube materials Cold sterilization results shown are for short duration 10 minute soak periods reactions may differ with extended contact Refer to Appendix A of this manual for information about specific solutions
25. assume that some fluid escaped the container or rotor Apply appropriate decontam ination procedures to the centrifuge rotor and accessories You can recover labware from most J series rotors while the rotor or yoke remains in the centrifuge Rotor buckets or carriers can be removed from the rotor yoke then unloaded on a lab bench or table or under a protective hood Blood bags must always be removed from blood bag cups outside of the centrifuge Using Tubes Bottles and Accessories Youcan remove the lid from most fixed angle rotors and extract the tubes or bottles using a removal tool specified in the applicable rotor manual I gt NOTE Vertical tube rotors cannot be unloaded inside the centrifuge The rotor must be removed from the centrifuge and placed in a rotor vise to loosen the tube cavity plugs Sample recovery depends on the type of labware used the compo nent s isolated and the analysis desired The Beckman Universal Fraction Recovery System 343890 can be useful when recovering sample from tubes see publication L5 TB 081 CAPPED TUBES The usual methods of recovering supernatants or pellets include decanting or withdrawing the gradient and scraping pellets from the tube bottom Remove tube caps carefully to avoid sample mixing e ftubes will be reused scrape pellets out with a plastic or wooden tool scratches on tube interiors caused by abrasive or sharply pointed tools can result in tube f
26. be recovered from most fixed angle rotors while the rotor remains in the centrifuge You can remove the lid and extract the tubes or bottles using the removal tool specified in the applicable rotor manual If the rotor is left in the centrifuge between runs be sure that it is securely tied down before each run Remove the rotor regularly and clean the drive spindle assembly 2 To remove the rotor lift it straight up and off the drive spindle IB NOTE If the rotor sticks to the drive spindle screw the short end of the rotor lifting handle assembly into the threaded opening to force the rotor off of the drive spindle hub Lubrication of the centrifuge drive spindle hub with Spinkote should prevent the rotor from sticking on all centrifuges except Avanti J series Avanti J series centrifuges have Delrin rings on the spindle hubs to prevent sticking and do not require lubrication Remove spacers tubes and bottles with the appropriate removal tool Refer to Section 3 USING TUBES BOTTLES AND ACCESSORIES for sample recovery methods J Series Rotors and Tubes J N DESCRIPTION J Series Rotors and Tubes 5 Using Swinging Bucket Rotors This section contains instructions for using swinging bucket rotors in J series centrifuges In addition to these instructions observe proce dures and precautions provided in the applicable rotor and centrifuge manuals Refer to Section 2 for tube selection information a
27. before the lid is attached or removed 3 Aerosolve cannisters are designed to minimize the possibility of aerosol and fluid leakage during centrifugation 3 4 J Series Rotors and Tubes Using Tubes Bottles and Accessories FILLING OPEN TOP TUBES OPEN TOP POLYALLOMER TUBES Open top polyallomer tubes are used in swinging bucket and fixed angle rotors Swinging Bucket Rotors Fill all opposing tubes to the same level Thinwall Tubes Fill to within 2 or 3 mm of the top for proper tube wall support Thickwall Tubes Fill at least half full Fixed Angle Rotors Fill all opposing tubes to the same level Thinwall Tubes Must be completely filled liquid and cap for support of the tube wall is critical e Thickwall Tubes Can be partially filled and centrifuged as indicated in the applicable rotor manual Speed reductions may be required for these partially filled tubes For greater fill volumes and faster speeds tube caps should be used Refer to the rotor manual for fill volumes and speed limitations OTHER OPEN TOP TUBES Open top tubes of other materials can also be used in fixed angle and swinging bucket rotors Vertical tube rotors use only Quick Seal tubes Fill these tubes as indicated below J Series Rotors and Tubes 3 5 Using Tubes Bottles and Accessories 3 6 Polycarbonate Thickwall polycarbonate tubes can be centrifuged partially filled Observe maximum rotor speeds and fi
28. depending on the need for transparency LABWARE TYPES I gt NOTE Tubes made of cellulose nitrate were formerly popular for various separations particularly rate zonal separations Beckman Coulter discon tinued the use of cellulose nitrate for tube manufacture in 1980 due to inconsistent physical properties inherent in the material If you currently have cellulose nitrate tubes dispose of them Consult your laboratory safety officer for proper disposal procedures J Series Rotors and Tubes ms Tubes Bottles and Accessories POLYALLOMER TUBES Polyallomer is a copolymer of ethylene and propylene Polyallomer tubes are translucent or transparent in appearance depending on wall thickness and are nonwettable although some polyallomer tubes can be chemically treated to make them wettable Polyallomer tubes have good tolerance to all gradient media including alkalines They perform well with most acids many bases many alcohols DMSO and some organic solvents Several types of polyallomer tubes are available Open Top Polyallomer Tubes Thinwall open top tubes are used in swinging bucket and fixed angle rotors In swinging bucket rotors thinwall tubes should be filled to within 2 or 3 mm of the tube top for proper tube support Caps are usually required in fixed angle rotors Thinwall tubes are designed for one time use and should be discarded after use Thickwall open top tubes offer the convenience of centrifuging
29. disks until the height of the assembly is nearly as tall as the tubes you will be using If the height of the disks is very tall you may have to push the bail into the grooves of the top disks by hand Remove or add disks to the bail to accommodate shorter or longer tubes If the tubes fit too snugly in the adapter s rubber base apply a light film of dusting power such as talcum powder to prevent the tubes from sticking J Series Rotors and Tubes 3 15 Using Tubes Bottles and Accessories USING AEROSOLVE CANNISTERS Aerosolve Cannister To tighten turn the lid from amp to B Tightening down the lid more than this will make it difficult to remove Aerosolve cannisters can be used in the JS 4 3 rotor to minimize aerosol leakage and liquid spills from rotor buckets during centrifuga tion Each cannister can hold a variety of tube sizes in tube racks that are specifically designed to fit in the cannisters The cannister can also be used as a 500 mL wide mouth bottle A WARNING When centrifuging hazardous materials always open cannisters in an appropriate hood or biological safety cabinet 1 Inspect cannister assemblies before use Do not use damaged components 2 Before placing the cannister in a bucket remove the bucket cover O ring seated on the ledge inside the bucket If this O ring is not removed a vacuum will be created between the bucket and cannister that will make removing the cannist
30. filled Plastic spacers have been tested for centrifugation between 2 and 25 C If spacers are centrifuged at temperatures significantly greater than 25 C deformation of the spacer and tube may occur ADAPTERS Many rotors can accommodate a variety of tube sizes by using adapters that line the tube cavity or bucket Adapters are fabricated of several different kinds of materials depending on the rotor and the tube to be used in them Some of the common materials are Delrin Noryl Ultem polyethylene rubber polypropylene and glass filled or foamed polypropylene Tubes or bottles used with adapters can be filled and capped if applicable according to the type of tube and the design of the rotor being used BOTTLE ADAPTERS Bottles are often supported during centrifugation in bottle adapters that fit inside the rotor buckets or cavities The adapters are usually Bottle ribbed for strength and support a variety of bottle sizes To prevent the bottles from stretching or breaking a plastic sleeve or adapter must be used around each Beckman Coulter 1 liter bottle during centrifugation in J6 series rotors In other rotors if the bottles fit snugly in the buckets the adapters are not required Refer to the applicable rotor manual Adapter 2 Delrin is a registered trademark of E I Du Pont de Nemours amp Company 3 Noryl and Ultem are registered trademarks of GE Plastics J Series Rotors and Tubes 2 9 Tubes Bott
31. if the rotor is forced sideways or dropped onto it Install the rotor by centering it over the spindle and carefully lowering it straight down 1 Carefully lower the rotor straight down onto the drive spindle 2 Rotate the rotor by hand until the drive pins seat on the drive spindle hub e In older model centrifuges be sure the pins in the rotor drive hole are located in the grooves of the drive spindle hub In newer model centrifuges be sure the pins in the rotor drive hole are not sitting on top of the teeth on the drive spindle hub 6 5 Using Vertical Tube and Rack Type Rotors A CAUTION The pins located in the rotor hub must be seated correctly on the centrifuge drive spindle Running a rotor that is not seated properly may result in rotor failure 3 Secure the rotor to the drive spindle hub with the rotor tie down assembly REMOVAL AND SAMPLE RECOVERY A CAUTION If disassembly reveals evidence of leakage you should assume that some fluid escaped the container or rotor Apply appropriate decon tamination procedures to the centrifuge and accessories 1 Remove the rotor tie down assembly 2 Lift the rotor straight up and off the drive spindle If the rotor sticks to the drive spindle a rotor removal tool may be used gt NOTE Lubrication of the centrifuge drive spindle hub with Spinkote should prevent the rotor from sticking on all centrifuges except Avanti J series N kotor Vise Avanti J seri
32. in the rotor bucket and simplify processing since they eliminate the need to remove buckets after each run They also minimize clean up downtime if a bag breaks simply remove the cup and resume the run Refer to the applicable rotor manual for blood bag cups used with each rotor Table D 2 lists blood bank methods that can be used for separating components in a variety of J6 series centrifuges TIPS FOR OPTIMUM CENTRIFUGATION RUNS Centrifugation generates high speeds causing rotor heads and buckets to develop a gravity force of thousands of pounds Observing the following tips will ensure safe and efficient operation N WARNING Handle body fluids with care because they can transmit disease No known test offers complete assurance that body fluids are free of micro organisms Some of the most virulent Hepatitis B and C and HIV I V viruses atypical mycobacterium and certain systemic fungi further emphasize the need for aerosol protection Never lift the centrifuge door while the instrument is running Keep metal clips needle holders and sealed tube ends away from blood bags Load opposing cups with equal weight to ensure safety optimum run efficiency and long rotor life Use weighted rubber disks for balancing e Load filled bags towards the outside wall of the bucket away from the centrifuge drive spindle Place the ADSOL bag between the blood bag and the plasma bag D 6 J Series Rotors and Tubes
33. into the rotor Opposing tubes must be filled to the same level with liquid of the same density Refer to ROTOR BALANCE in Section 1 Refer to the applicable centrifuge instruction manual for detailed operating information For low temperature runs precool the rotor in the centrifuge or in a refrigerator before use especially before short runs to ensure that the rotor reaches the set temperature To ensure that the rotor reaches the required temperature during centrifugation some temperature compensation may be required because of the mass of these rotors Refer to Appendix B or to the applicable rotor manual for tables listing temperature compensation units for various rotors If you are using an Avanti J series centrifuge select the rotor number e If you are using a microprocessor controlled J2 or J6 series centri fuge enter the rotor code if the JA 10 rotor is used for example enter code 10 L CAUTION The centrifuge drive spindle can be bent or broken if the rotor is forced sideways or dropped onto it Install the rotor by centering it over the spindle and carefully lowering it straight down 1 Carefully lower the rotor straight down onto the drive spindle Rotate it by hand until the drive pins seat on the drive spindle hub J Series Rotors and Tubes Rotor Drive Pins 2 Hub Grooves 4 LC SERIE NS Drive Spindle Assembly Older Model Centrifuges Teeth 2 Psy N Drive Spindle Assembly Newer Mod
34. must contain either a bucket or a microtiter plate carrier during a run Consult the applicable centrifuge instruction manual for operating instructions USING MICROTITER PLATE CARRIERS Micro Plus Carriers Anodized aluminum microtiter plate carriers can be installed on the pivot pins in place of the buckets normally used with some swinging bucket rotors Each carrier allows centrifugation of up to three 96 well microtiter plates For complete information about the carriers see publication GS6 TB 011 which accompanies the Micro Plus carriers or publication J6 TB 009 which accompanies the J6 series carriers Microplates will break if g forces are too high Rotor speed must be reduced when microplate carriers are used If microplate carriers and buckets are centrifuged in the same run run speed must be reduced to the speed allowable for the microplates Refer to the applicable rotor manual for allowable run speeds If only two carriers are run they must be installed opposite each other in the rotor and the remaining positions on the yoke must be filled with either buckets or other carriers they need not be loaded to prevent rotor imbalance See SYMMETRIC AND BALANCED LOADING above 1 To prevent microtiter plate breakage during centrifugation place the flexible plastic pad ridged side up into the flat indented area of the blue base see Figure 5 4 2 Place the plate s on top of the pad being careful not to spill
35. partially filled tubes without tube caps in fixed angle and swinging bucket rotors Because the solution reorients during centrifugation the maximum partial fill volume depends on the tube angle For greater fill volumes use tubes with caps Refer to the applicable rotor J manual for fill volumes and speed reduction requirements Thickwall tubes are reusable Quick Seal Polyallomer Tubes g Max Heat sealed Quick Seal tubes can be used in some fixed angle rotors hee oe and in the JS 24 series rotors they must be used in the JV 20 vertical tube rotor Single use Quick Seal tubes are a convenient form of x lil sealable tube they are especially useful for the containment of radio active or pathogenic samples There are two Quick Seal tube designs dome top and bell top The bell top simplifies removal of materials that float during centrifugation A Dome top tubes hold more volume than their bell top equivalents Dome Top Bell Top Detailed information about Quick Seal tubes is contained in publication IN 181 2 4 J Series Rotors and Tubes POLYCARBONATE TUBES C POLYPROPYLENE TUBES POLYETHYLENE TUBES J Series Rotors and Tubes Tubes Bottles and Accessories Polycarbonate is tough rigid nonwettable and glass like in appear ance Polycarbonate tubes are reusable and can be used with or without caps in fixed angle rotors and at least half full in swinging bucket rotors Speed reduction may be required in
36. saline adenine glucose minitol an additive used as a stabilizer for red cells that extends red cell life in CPD the settling out of particles from a suspension in the earth s field of gravity in the centrifuge this process is accelerated and the particles move away from the axis of rotation sedimentation velocity per unit of centrifugal force dr 1 x d ar S sodium dodecyl sulfate an ionic detergent used in cell lysis and solubilizing of proteins a large group of silicone elastomers used in various accessories should be used at temperatures between 59 and 232 C 75 and 450 F short centrifugation run 3 to 5 minutes at low g forces 2000 to 3000 x g at ambient temperature used to keep small cells or platelets in suspension while the larger cellular components sediment used to obtain platelet rich plasma and red blood cell concentrate from whole blood Beckman Coulter concentrated rotor cleaning solution recommended because it is a mild solution that has been tested and found effective and safe for Beckman Coulter rotors and accessories E 5 Glossary of Terms SpinkoteTM Beckman Coulter lubricant for metal to metal contacts sucrose a sugar not a self forming gradient used in rate zonal separations generally used in separating RNA subcellular organelles and cell membranes supernatant the liquid above the sedimented material following centrifugation Svedberg unit S a unit of sedimentation velocity
37. some rotors if the tubes are not completely filled Although polycarbonate tubes may be autoclaved doing so greatly reduces the usable life of these tubes Cold sterilization methods are recommended Washing with alkaline detergents can cause failure Crazing the appearance of fine cracks in the tube is the result of stress relaxation and can affect tube performance These cracks will gradually increase in size and depth becoming more visible Tubes should be discarded before cracks become large enough for fluid to escape These tubes have good tolerance to all gradient media except alkalines pH greater than 8 They are satisfactory for some weak acids but are unsatisfactory for all bases alcohol and other organic solvents Polypropylene tubes are translucent in appearance and are reusable unless deformed during centrifugation or autoclaving These tubes have good tolerance to gradient media including alkalines They are satisfactory for many acids bases and alcohols but are unsatisfac tory for most organic solvents They can be used with or without caps in fixed angle rotors Speed reduction is sometimes required with these tubes if run with less than full volume refer to your rotor manual Polyethylene tubes are translucent or transparent and have a good tolerance for use with strong acids and bases They are reusable but cannot be autoclaved In swinging bucket rotors they are used without caps and with or without caps i
38. stress of centrifugation or with extended contact or temperature variations To prevent failure and loss of valuable sample ALWAYS TEST SOLUTIONS UNDER OPERATING CONDITIONS BEFORE USE N WARNING Do not use flammable substances in or near operating centrifuges 2 2 J Series Rotors and Tubes Tubes Bottles and Accessories LABWARE MATERIAL COMPATIBILITY WITH SOLVENTS AND SAMPLE The chemical compatibility of tube or bottle materials with the gradient forming medium or other chemicals in the solution is an important consideration Although neutral sucrose and salt solutions cause no problems alkaline solutions cannot be used in Ultra Clear tubes or in polycarbonate tubes and bottles Polycarbonate and Ultra Clear tubes are incompatible with DMSO sometimes used in the preparation of sucrose gradients for sedimentation of denatured DNA GRADIENT FORMATION AND FRACTIONATION Consideration should be given to gradient formation and fractionation when choosing a tube for a density gradient run If the bands or zones formed during centrifugation are indistinct they may not be visible through a translucent material such as polyallomer If optimum band visualization is important Ultra Clear polycarbonate or cellulose propionate tubes should be used Whenever collection of bands or zones must be done by slicing or puncturing the tube a thin flexible tube wall is required Ultra Clear or polyallomer tubes should be used in these cases
39. t is time in hours k is the clearing factor of the rotor and s is the sedimentation coefficient in Svedberg units S citrate phosphate dextrose anti coagulant and preservative citrate phosphate dextrose citrate citric acid adenine anti coagulant and preservative a precipitate such as cryoglobulin or antihemophilic factor VIII that results from cooling cesium chloride a high density salt used in solution in isopycnic separations to separate particles based on their density cesium sulfate a salt similar to CsCl that will form its own gradient in solution thermoplastic material acetal homopolymer used for most tube adapters Delrin is a registered trademark of E I Du Pont de Nemours amp Company mass per unit volume a centrifugal separation process based on differences in particle densities a centrifugal separation process based on differences in particle sizes ethylene propylene rubber used for O rings and pad adapters should be used at temperatures between 57 and 120 C 70 and 250 F see RBC red blood cells J Series Rotors and Tubes ethidium bromide fixed angle rotor granulocytes hard spin HDPE isopycnic LDPE leukapheresis leukocytes lymphocyte maximum volume meniscus NaCl neoprene J Series Rotors and Tubes Glossary of Terms a fluorescent intercalating orange dye used commonly in the separation of DNA and in gel electrophoresis a rotor in which the tubes a
40. the rotor before placing the rotor into the centrifuge and to remove the rotor from the centrifuge with the lid attached N CAUTION Always loosen the rotor knob before loosening the daisy knob to avoid jamming the knobs Other rotors are secured to the centrifuge drive spindle hub by a tie down screw A torque bar is supplied with the rotor to provide leverage to securely fasten the rotor Rotors are specifically designed to withstand a maximum load that is volume and density of the rotor contents at rated speed At greater speeds or at rated speeds with heavier loads rotors are subject to failure nJ series centrifuges with analog controls the rotor speed is limited by the physical properties of the rotor Friction created by the air in the centrifuge chamber interacting with the rotor surfaces during centrifugation in most cases prevents rotors from exceeding their rated speeds nmicroprocessor controlled J2 and J6 series centrifuges internal circuitry monitors the rotor speed and prevents a rotor from exceeding its rated speed The rotor entry code listed in the appli cable rotor manual sets the allowable speed n Avanti J series centrifuges an electronic recognition system identifies the rotor thereby limiting speed to the rated speed of the rotor At rated speeds with heavier loads rotors are subject to failure It is the operator s responsibility to limit rotor speed when centrifuging dense solutions or
41. the tube to float and collapse Ensure that tubes and tube cavities or buckets are dry before inserting the tubes Reagent used that attacks the tube material Refer to Appendix A for chemical compatibilities of tube material and chemicals Tubes run above their rated speed Refer to the applicable rotor manual for maximum speeds Bottle leakage bottles with cap assemblies Moisture or lubrication on cap or sealing surface Ensure that the O ring plug and bottle lip are dry and free of lubrication before use O ring or gasket damaged or defective Replace the O ring or gasket Cap not tightened sufficiently Tighten cap securely Sealing surface of the bottle is not smooth Replace bottle Threaded caps without inserts or O rings Tube too full these are not as liquid tight as cap assemblies therefore the meniscus must be kept lower to prevent leakage Bottle leakage uncapped bottles Bottle too full the meniscus must be kept lower to prevent leakage Refer to the rotor manual for fill volumes and speed reductions Bottle damage 7 14 Fill volume too low to provide tube wall support Refer to the rotor manual for fill volumes and speed reduction Moisture between the bottle and the cavity or bucket can cause the bottle to float and collapse Ensure that bottles and cavities or buckets are dry before inserting them Reagent used that attacks the bottle material Refer to Appendix A for chemical compatibilitie
42. to Beckman Coulter Inc 1050 Page Mill Road Palo Alto CA 94304 Attention Returned Goods Customers located outside the United States should contact their local Beckman Coulter office J Series Rotors and Tubes J i Care and Maintenance DIAGNOSTIC HINTS Some of the more common operating problems experienced in centrifugation are listed below with suggestions for their solutions Contact your Beckman Coulter Field Service representative if a problem cannot be corrected I gt NOTE Use only the labware listed in the applicable rotor manual SYMPTOM POSSIBLE CAUSE AND SUGGESTED ACTION Rotors Severe vibration Rotor imbalance To balance the rotor load fill all opposing tubes to the same level with liquid of the same density Weight of opposing tubes must be distributed equally Place tubes in a fixed angle or vertical tube rotor symmetrically as illustrated in Section 1 Figure 1 7 Detailed information about balancing swinging bucket rotors is contained in Section 5 Speed selected is within the rotor s critical speed range Select a speed outside the critical speed range Refer to the applicable rotor manual for critical speed range Rotor improperly tied down Make sure the rotor is properly secured to the drive spindle hub before centrifugation If the rotor is left in the centrifuge between runs tighten the tie down device before each run Swinging bucket rotor Mishooked bucket loose bucket
43. trays which are then loaded into carriers at a resting angle During centrifugation the carriers swing out to a completely horizontal position Refer to Section 6 for specific information about the use of rack type rotors J Series Rotors and Tubes Rotors At Speed At Rest in Rotor At Rest Outside Rotor Fixed Angle Rotors Pathlength Swinging Bucket Rotors Pathlength Vertical Tube Rotors fmin fmax I Pathlength Figure 1 2 Particle Separation in Fixed Angle Swinging Bucket and Vertical Tube Rotors Dark gray represents pelleted material light gray is floating components and bands are indicated by black lines J Series Rotors and Tubes 1 9 Rotors PELLETING DIFFERENTIAL SEPARATION Pelleting separates particles of different sedimentation coefficients the largest particles in the sample traveling to the bottom of the tube or bottle first Differential centrifugation is the successive pelleting of particles of decreasing sedimentation velocities using increasingly higher forces and or long run times The relative pelleting efficiency of each rotor is measured by its k factor clearing factor ic Ininax Tmin 103 CETT 1 n2 3600 where c is the angular velocity of the rotor in radians per second 21 RPM 60 or 0 10472 x rpm Fax is the maximum radius and ry is the minimum radius After substitution 2 rpm This factor can be used in the following equation to estimate the ti
44. two quarts of Solution 555 and brushes that will not scratch rotor surfaces 2 Rinse thoroughly with water 3 Air dry the body or buckets upside down Do not use acetone to dry rotors Wipe clean the O rings or gaskets regularly lubricate after cleaning Replace them about twice a year or as required Frequently clean all surfaces that contact O rings Regularly clean the threads of the rotor lid handle buckets cavities and so on with a nonmetal brush and a small amount of concentrated detergent then rinse and dry thoroughly Lubricate the threads as directed under LUBRICATION below J Series Rotors and Tubes Care and Maintenance Approximately once a week or every 250 runs clean the pins and bucket pin sockets of swinging bucket rotors to prevent buildup of residues After cleaning lubricate as described under LUBRICATION below A CAUTION Do not use acetone MEK methylethylketone chloroform cyclohexane or organic solvents on carbon fiber cannisters at any time These substances will damage the epoxy resin surface material DECONTAMINATION Rotors contaminated with radioactive or pathogenic materials must be decontaminated following appropriate laboratory safety guide lines and or other regulations III NOTE Strong bases and or high pH solutions can damage aluminum rotors and components Ifa rotor and or accessories becomes contaminated with radioac tive material it should be decontamin
45. virus RNA Bacteriophage T5 DNA 60 Bacteriophage T2 amp T4 DNAs Broad bean mottle Ribosomes Poliomyelitis 200 Tobacco mosaic Equine encephalitis Polysomes Viruses Rous sarcoma 800 Feline leukemia Bacteriophage T2 Microsomes Subcellular Particles Plasma membranes Mitochondria 100 000 Figure 1 3 Sedimentation Coefficients in Svedberg Units for Some Common Biological Materials J Series Rotors and Tubes 1 11 Rotors The centrifugal force exerted at a given radius in a rotor is a function of the rotor speed The nomograms for J2 series and J6 series centri fuges in Figures 1 4 and 1 5 allow you to determine relative centrifugal field RCF for a given radius and rotor speed In Avanti J series centrifuges the RCF is calculated automatically by the centri fuge software Run times can be shortened by using partially filled thickwall polyal lomer and polycarbonate tubes The short pathlength means less distance for particles to travel in the portion of the tube experiencing greatest centrifugal force and hence shortened run times The k factors for half filled tubes can be calculated by using an approximate Fmax and ry in k factor equation 1 21 000 180 90 000 50 EDD 50 000 150 15 000 10 000 10 000 100 5 000 90 i 5 000 70 1 000 60 500 50 200 2 000 B 100 50
46. when using heavy containers refer to ALLOW ABLE RUN SPEEDS below Rotors ALLOWABLE RUN SPEEDS Under some conditions the maximum allowable speed of the rotor indicated by the rotor name must be reduced to ensure that neither oor the rotor nor the labware are overstressed during centrifugation SPEED RPM RCF Dense Solutions When using dense solutions gt 1 2 g mL in J2 series rotors determine maximum run speed using the following square root reduction formula P reduced run speed maximum rated speed 7 B where p 4 is the maximum permissible density of the tube contents for a particular rotor from the rotor manual and pp is the actual density of the tube contents to be centrifuged When using dense solutions in J6 series rotors determine maxi mum run speed using the following square root reduction formula reduced run speed 2 maximum rated speed I 8 where A is 2500 grams for JS rotors or 1500 grams for the JR 3 2 rotor and B is the weight in grams of a total load bucket with adapter and sample bucket with blood bag cup and filled blood bag tray with racks tubes and sample I gt NOTE The maximum speed for Avanti J or J2 series rotors in J6 series instruments is 6000 rpm with solutions of density no greater than 2 0 g mL Solutions of density greater than 2 0 g mL should not be centrifuged Critical Speed Range The critical speed range of a rotor is the range of speeds in which dur
47. 0 I centrifuge only In Avanti J 25 series and J 20 series centrifuges the maximum speed for these rotors is 10 000 rpm J Series Rotors and Tubes Rotors Table 1 1 Rotors Used in Beckman Coulter J Series Centrifuges continued Rotors in parentheses are no longer manufactured Swinging Bucket continued JS 4 3 3 liters 4 300 4 220 16 635 JS 4 2 6 liters 4 200 5 020 11 502 JS 4 2A 6 liters 4 200 5 020 11 502 JS 4 2SM 6 quad 4 200 4 900 blood bags JS 4 2SMA 6 quad 4 200 4 900 blood bags JS 4 20 4 liters 4 000 4044 15 298 JS 3 4A 1250 7 5 liters 3 400 3 370 18 066 JS 3 0 6 liters 3 000 2 560 22 598 JS 2 9 6 liters 2 900 2 500 24 400 Vertical Tube and Rack Type JV 20 312 mL 20 000 41 619 206 JR 3 2 320 mL Zonal and Continuous Flow see applicable rotor manual for rotor description and use JCF Z 100 L hr 20 000 39 900 X X X X X X HF seal assembly 45 L hr SF seal assembly JE 5 0 1000 mL 5 000 4 700 X X X X JE 6B 100 mL 6 000 5 080 X X X X X a Maximum speed in an Avanti J E centrifuge is 21 000 rpm 18 000 rpm at 2 C at 35 C ambient and 95 percent humidity b When a JA 18 1 rotor is used in a J2 HC centrifuge derate the rotor as follows when the 45 adapters are used do not run the rotor above 15 000 rpm when 25 adapters are used do not run the rotor above 16 000 rpm Maximum speed in an Avanti J E for the roto
48. 00 2 500 X X Swinging Bucket JS 24 38 231 mL 240004 103 900 334 X X X JS 24 15 90 mL 24000 110500 376 X X X JS 13 1 300 mL 13 000 26 500 1841 X X X X X X X JS 7 5 200 mL 7 500 10 400 1090 X X X X X X JS 5 9 384 mL 5 900 6 570 X X JS 5 3 691 mL 5 300 6 130 X X JS 5 2 4 liters 5 200 6 840 9 051 X X JS 5 0 9 liters 5 000 7 480 9171 X Continued a Maximum speed in an Avanti J E centrifuge is 21 000 rpm 18 000 rpm at 2 C at 35 C ambient and 95 percent humidity b When a JA 18 1 rotor is used in a J2 HC centrifuge derate the rotor as follows when the 45 adapters are used do not run the rotor above 15 000 rpm when 25 adapters are used do not run the rotor above 16 000 rpm Maximum speed in an Avanti J E for the rotor with magnets without magnets maximum is 14 000 rpm Maximum speed at 2 C in a 50 Hz centrifuge is 14 000 rpm 4 Maximum speed in an Avanti J E for the rotor with magnets without magnets maximum is 13 000 rpm Maximum speed at 2 C in a 50 Hz centrifuge is 15 000 rpm Maximum speed in an Avanti J series centrifuge Maximum speed in a J2 series centrifuge is 14 000 rpm f Maximum speed in an Avanti J E for rotor without magnets is 6 300 rpm Maximum speed for rotor in an Avanti J E centrifuge is 6 300 rpm h The JS 24 38 and JS 24 15 rotors can achieve 24 000 rpm in an Avanti J 3
49. 000 e eee ee 3 8 JLA 8 1000 and JLA 9 1000 Bottle Cap Closures 3 8 Filling and Loading Cups in the JS 5 0 Rotor 3 8 Filling and Sealing Quick Seal Tubes 004 3 9 Method A With the Seal Guide 0 3 12 Method B Without the Seal Guide 3 13 Capping Multiwell Titer Plates 040 3 14 Using A daptets 4 camen cabe tas REY Eg hia wethe da 3 14 Using Solid Multitube Adapters 004 3 15 Using Modular Disk Multitube Adapters 3 15 Using Aerosolve Cannisters 0 0 0 cece eee eee eese 3 16 Using Cannisters as Wide Mouth Bottles 3 17 Using Cannisters with Tube Racks 044 3 17 Using Blood Bag Cups eseseeee eee eee 3 18 sample Recovery cese fess anes eke IRR ERR oo 3 19 Capped Tubes conc oe eda RC UV d ud 3 20 J5 5 0 Cups i ect aestate S tes takes d 3 20 Quick Seal Tubes 0 0 cece ee 3 21 Making Ultra Clear Tubes Wettable 0 3 22 J Series Rotors and Tubes J Series Rotors and Tubes SECTION 4 SECTION 5 SECTION 6 USING FIXED ANGLE ROTORS Description 12d e eatem eee Le Re e o ble Y ERR a UM E OE ALORS Tubes and Bottles llelee eee Rotor Preparation and Loading 0 2 0 0 scene eee Prerun Safety Checks cc eee eee esee ens Rotor Preparation 0 0 eee eee eee eee
50. 0297 288082 SINGAPORE Beckman Coulter Singapore Pte Ltd 116 Changi Road Unit 03 01 02 Singapore 419718 Telephone 65 6339 3633 Fax 65 6336 6303 SOUTH AFRICA SUB SAHARAN AFRICA Beckman Coulter Stand 1A Primegro Park Tonetti Street 1685 Halfway House Johannesburg Republic of South Africa Telephone 27 11 805 2014 5 Fax 27 11 805 4120 e mail beckman intekom co za SPAIN Beckman Coulter Espa a S A C Caleruega 81 28033 Madrid Spain Telephone 34 91 3836080 Fax 34 91 3836096 email bioresearch espana 9 beckman com SWEDEN Beckman Coulter AB Archimedesvaegen 7 Box 111 56 SE 168 11 Bromma Sweden Telephone 0 8 564 85 900 Telefax 0 8 564 85 901 SWITZERLAND Beckman Coulter International S A 22 Rue Juste Olivier Case Postale 301 303 CH 1260 Nyon Switzerland Telephone 0800 850 810 Fax 0848 850 810 TAIWAN Beckman Coulter Taiwan Inc Taiwan Branch 8th Floor 216 Tun Hwa South Road Section 2 Taipei 106 Taiwan Republic of China Telephone 886 2 2378 3456 Fax 886 2 2377 0408 TURKEY Beckman Coulter Ltd E 5 Yanyol Faith Cad 81410 Soganlik Kartal Istanbul Turkey Telephone 90 216 309 1900 Fax 90 216 309 0090 UNITED KINGDOM Beckman Coulter United Kingdom Ltd Oakley Court Kingsmead Business Park London Road High Wycombe Buckinghamshire HP11 1JU England U K Telephone 01494 441181 Fax 01494 447558 e mail beckman uk beckman com
51. 1 1 2164 1 3539 24 291 9 1 734 1 720 1 4012 57 980 4 5 823 1 2275 1 3550 25 306 9 1 823 1 741 1 4032 58 1009 8 5 998 1 2387 1 3561 26 322 1 1 913 1 763 1 4052 59 1040 2 6 178 1 2502 1 3572 27 337 6 2 005 1 7846 1 4072 60 1070 8 6 360 1 2619 1 3584 28 353 3 2 098 1 808 1 4093 61 1102 9 6 550 1 2738 1 3596 29 369 4 2 194 1 831 1 4115 62 1135 8 6 746 1 2858 1 3607 30 385 7 2 291 1 856 1 4137 63 1167 3 6 945 1 298 1 3619 31 402 4 2 390 1 880 1 4160 64 1203 2 7 146 1 311 1 3631 32 419 5 2 492 1 9052 1 4183 65 1238 4 7 355 1 324 1 3644 33 436 9 2 595 Computed from the relationship p gt 10 2402 nD 5 12 6483 for densities between 1 00 and 1 37 and p 5 10 8601 nD25 13 4974 for densities above 1 37 Bruner and Vinograd 1965 Divide by 10 0 to obtain 96 w v Density data are from International Critical Tables J Series Rotors and Tubes C 3 Gradient Materials Table C 3 Density Refractive Index and Concentration Data Sucrose at 20 C Molecular Weight 342 3 Density Refractive 96 by mg mL of Density Refractive 96 by mg mL of g cm3 Index nD Weight Solution Molarity g cm3 Index nD Weight Solution Molarity 0 9982 1 3330 0 1 1463 1 3883 34 389 7 1 138 1 0021 1 3344 1 10 0 0 029 1 1513 1 3902 35 403 0 1 177 1 0060 1 3359 2 20 1 0 059 1 1562 1 3920 36 416 2 1 216 1 0099 1 3374 3 30 3 0 089 1 1612 1 3939 37 429 6 1 255 1 0139 1 338
52. 40 30 30 1 000 20 25 15 750 Radial Distance Relative Centrifugal Field Speed mm x g rpm Figure 1 4 Nomogram for J2 Series Centrifuges Align a straightedge through known values in two columns read the figure where the straightedge intersects the third column 1 12 J Series Rotors and Tubes 350 15 000 6 000 10 000 5 000 300 5 000 4 000 250 2 000 POE 200 1 000 2000 180 160 500 14 z 200 J20 1 000 100 100 50 90 500 80 20 70 10 60 5 50 2 200 Radial Distance Relative Centrifugal Field Speed mm x g rpm Figure 1 5 Nomogram for J6 Series Centrifuges Align a straightedge through known values in two columns read the figure where the straightedge intersects the third column ISOPYCNIC SEPARATIONS A sedimentation equilibrium or isopycnic method separates particles on the basis of particle buoyant density Each component in the sample travels through the gradient until it reaches an equilibrium position Particle velocity due to differences in density is given in the following expression d p p _ p c v ae E 6 Rotors RATE ZONAL SEPARATIONS where v sedimentation velocity dr dr d particle diameter Pp particle density Pe solution density u viscosity of liquid media g standard acceleration of gravity At equilibrium py p is zero and particle velocity is therefore zero The gradient may be preformed before the run or generated during centrifugation For gradients f
53. 49 1 16053 1 15996 1 1696 1 14596 1 15526 1 15605 1 1652 1 1401 20 1 17580 1 18107 1 18112 1 1913 1 16469 1 17554 1 17657 1 1864 1 1580 22 1 19679 1 20362 1 20305 1 2137 1 18396 1 19650 1 19781 1 2083 1 1763 24 1 21849 1 22634 1 22580 1 2375 1 20379 1 21817 1 21980 1 2309 1 1952 26 1 24093 1 24990 1 24942 1 2643 1 22421 1 24059 1 24257 1 2542 1 2146 28 1 26414 1 27435 1 27395 1 24524 1 26380 1 26616 1 2782 1 2346 30 1 28817 1 29973 1 29944 1 26691 1 28784 1 29061 1 3028 1 2552 35 1 35218 1 36764 1 36776 1 32407 1 35191 1 35598 1 3281 1 2764 40 1 42245 1 44275 1 44354 1 38599 1 42233 1 42806 45 1 49993 1 52626 1 52803 1 45330 1 50010 1 50792 50 1 58575 1 61970 1 62278 1 52675 1 58639 1 59691 55 1 68137 1 72492 1 68254 1 69667 60 1 78859 1 80924 65 1 90966 1 93722 J Series Rotors and Tubes C 5 Appendix D Blood Component Separation This Appendix provides a basic overview of blood separation proce dures using Beckman Coulter J series centrifuges BLOOD BANK COLLECTION OVERVIEW J Series Rotors and Tubes Blood is composed of plasma red blood cells RBC white blood cells WBC and platelets Approximately 40 to 45 percent of this volume is made up of red blood cells most of the remainder is plasma a watery substance that contains vital substances including hormones and proteins Most whole blood collected undergoes fract
54. 6 3900 6 0 6or7 1 4 5 5 5 JS 5 2 4 4100 6 0 6 1 4 5 5 5 Cryoprecipitated Plasma Hard Spin 4 JS 4 2 6 4200 7 2 Max Max Max Antihemophilic Cell free JS 4 2SM 6 4200 7 5 Max Max Max Factor frozen and JS 5 2 4 4500 7 1 Max Max Max then thawed at 4 C Times include acceleration and time at maximum speed only Deceleration time is not included Brake settings are estimated for a rotor fully loaded with 500 mL blood bags When using other bags brake settings should be increased to maintain comparable deceleration times 3 Several methods for preparation of platelet rich plasma and platelet concentrate are in current use The speed and time ranges given have been estimated to be comparable to conditions specified in the following sources American Association of Blood Banks Technical Manual p 359 7th ed washington 1977 Humphreys P Private communication Canadian Red Cross Society Toronto Ontario July 1977 Kahn R A Cossette I Friedman L I Transfusion 16 162 165 1976 Reiss R F Katz A J Transfusion 16 370 374 1976 Slichter S J Harker L A Transfusion 16 8 12 1976 J Series Rotors and Tubes D 5 Blood Component Separation Beckman Coulter has centrifuges rotors and accessories designed to fit the special needs of blood component processing Several rotors are available to accommodate single double triple and quad blood bags Blood bag cups rest
55. 8 4 40 6 0 119 1 1663 1 3958 38 443 2 1 295 1 0179 1 3403 5 50 9 0 149 1 1713 1 3978 39 456 8 1 334 1 0219 1 3418 6 61 3 0 179 1 1764 1 3997 40 470 6 1 375 1 0259 1 3433 7 71 8 0 210 1 1816 1 4016 41 484 5 1 415 1 0299 1 3448 8 82 4 0 211 1 1868 1 4036 42 498 5 1 456 1 0340 1 3464 9 93 1 0 272 1 1920 1 4056 43 512 6 1 498 1 0381 1 3479 10 103 8 0 303 1 1972 1 4076 44 526 8 1 539 1 0423 1 3494 11 114 7 0 335 1 2025 1 4096 45 541 1 1 581 1 0465 1 3510 12 125 6 0 367 1 2079 1 4117 46 555 6 1 623 1 0507 1 3526 13 136 6 0 399 1 2132 1 4137 47 570 2 1 666 1 0549 1 3541 14 147 7 0 431 1 2186 1 4158 48 584 9 1 709 1 0592 1 3557 15 158 9 0 464 1 2241 1 4179 49 599 8 1 752 1 0635 1 3573 16 170 2 0 497 1 2296 1 4200 50 614 8 1 796 1 0678 1 3590 17 181 5 0 530 1 2351 1 4221 51 629 9 1 840 1 0721 1 3606 18 193 0 0 564 1 2406 1 4242 52 645 1 1 885 1 0765 1 3622 19 204 5 0 597 1 2462 1 4264 53 660 5 1 930 1 0810 1 3639 20 216 2 0 632 1 2519 1 4285 54 676 0 1 975 1 0854 1 3655 21 227 9 0 666 1 2575 1 5307 55 691 6 2 020 1 0899 1 3672 22 239 8 0 701 1 2632 1 4329 56 707 4 2 067 1 0944 1 3689 23 251 7 0 735 1 2690 1 4351 57 723 3 2 113 1 0990 1 3706 24 263 8 0 771 1 2748 1 4373 58 739 4 2 160 1 1036 1 3723 25 275 9 0 806 1 2806 1 4396 59 755 6 2 207 1 1082 1 3740 26 288 1 0 842 1 2865 1 4418 60 711 9 2 255 1 1128 1 3758 27 300 5 0 878 1 2924 1 4441 62 788 3 2 303 1 1175 1 3775 28 312 9 0 914 1 2983 1 4464 62 804 9 2 351 1 1222 1 3793 29 325 4 0 951 1 3043 1 4486 63 821
56. DISINFECTION 121 C Goa ub INSPECTION When sterilization or disinfection is a concern consult your labora tory safety officer regarding proper methods to use While Beckman Coulter has tested the following methods and found that they do not damage the rotor or components no guarantee of sterility or disinfec tion is expressed or implied Rotors and most rotor components can be autoclaved at 121 C for up to an hour Remove the lid and place the rotor and or buckets in the autoclave upside down O rings and gaskets can be left in place on the rotor Ethanol 70 may be used on all rotor components including those made of plastic Bleach sodium hypochlorite may be used but may cause discoloration of anodized surfaces Use the minimum immersion time for each solution per laboratory standards Frequent and thorough inspection is crucial to maintaining a rotor in good operating condition Periodically at least monthly depending on use inspect the rotor especially inside cavities and buckets for rough spots cracks pitting white powder deposits on aluminum rotors frequently aluminum oxide or heavy discoloration If any of these signs are evident do not run the rotor Contact your Beckman Coulter repre sentative for information about the Field Rotor Inspection Program and the Rotor Repair Program 3 Flammability hazard Do not use in or near operating centrifuges 7 4 J Series Rotors and Tubes
57. Insert spacers and floating spacers as listed in the rotor manual to completely fill rotor cavities in use Floating Spacer 5 With the rotor in the rotor vise 332688 insert plugs over filled cavities only do not insert plugs in empty cavities Tighten the Nay plugs using the plug wrench provided 340632 6 4 J Series Rotors and Tubes OPERATION Rotor JV 20 INSTALLING THE ROTOR Rotor Drive Pins 2 m Grooves 4 Sus NS Drive Spindle Assembly Older Model Centrifuges Teeth 2 ore NS Drive Spindle Assembly Newer Model Centrifuges J Series Rotors and Tubes Using Vertical Tube and Rack Type Rotors Refer to the centrifuge instruction manual for detailed operating information For low temperature runs precool the rotor in the centri fuge or in a refrigerator before use especially before short runs to ensure that the rotor reaches the set temperature To ensure that the rotor reaches the required temperature during centrifugation some temperature compensation may be required because of the mass of these rotors Refer to Appendix B or to the rotor manual for tables listing temperature compensation units for various rotors If you are using an Avanti J series centrifuge select the rotor number If you are using a microprocessor controlled J2 or J6 series centrifuge enter the rotor code enter code 20 for the JV 20 rotor N CAUTION The centrifuge drive spindle can be bent or broken
58. JR IM 6 COULTER ROTORS AND TUBES For Beckman Coulter J2 J6 and Avanti J Series Centrifuges User s Manual my Cou yey Publishedby the Centrifuge Instrument Systems Development Center Of Beckman Coulter Inc Palo Alto California 94304 SAFETY NOTICE This safety notice summarizes information basic to the safe operation of the rotors and accessories described in this manual The international symbol displayed above is a reminder that all safety instructions should be read and understood before use or mainte nance of rotors or accessories When you see the symbol on other pages pay special attention to the safety information presented Also observe any safety information contained in applicable rotor and centrifuge manuals Observance of safety precautions will help to avoid actions that could cause personal injury as well as damage or adversely affect the performance of the centrifuge rotor tube system Chemical and Biological Safety If pathogenic toxic or radioactive samples are intended to be used it is the responsibility of the user to ensure that all necessary safety regulations guidelines precautions and practices are adhered to accordingly Ask your laboratory safety officer to advise you about the level of containment required for your application and about proper decontami nation or sterilization procedures to follow if fluids escape from their containers Observe all cautionar
59. JS 7 5 5 000 2 0 43 3 3 3 4 2 000 0 2 4 4 4 4 5 20 000 N N N N max max max 18 000 N N N max 10 10 9 act 15 000 N N 7 7 6 5 4 10 000 N N 2 2 1 0 0 Above 30 C ambient temperature this temperature may not be achieved at this speed N indicates that the rotor cannot achieve the desired temperature at this speed J Series Rotors and Tubes B 7 Temperature Compensation Tables B 8 JS 5 2 Table B 4 Temperature Compensation Settings for the J6 Centrifuges 5 200 4 000 3 000 2 000 and below Ratia ONO rotors ee 00 1 7 9 14 20 30 JSAM agp and 2 4 8 10 15 20 30 JS 4 2A below JS 4 2SMA All other all speeds 2 4 8 10 15 20 30 J Series Rotors and Tubes J Series Rotors and Tubes Appendix C Gradient Materials This Appendix contains reference information on commonly used gradient materials General instructions for filling and sealing tubes including gradient preparation are contained in Section 3 Gradient material selection depends on a number of factors including the type of separation to be performed Sucrose is used for rate zonal and isopycnic separations and cesium chloride is often used for isopycnic separations The basic requirement is that the gradient permit the type of separation Additional considerations in selecting a gradient material include the following Its density range should be
60. LA 10 500 8 000 1 1 0 2 2 3 1 5 000 0 1 2 1 4 3 1 13 000 N 10 10 10 10 10 9 11 000 N 8 8 8 7 6 5 med 8 000 6 5 4 4 3 3 N 5 000 4 3 1 1 1 1 N 7 500 4 3 1 1 1 2 3 JS 7 5 5 000 2 0 43 3 3 3 4 2 000 0 2 4 4 4 4 5 20 000 N N max max max max max JV 20 18 000 N N max max 10 10 9 15 000 N N 7 7 6 5 4 10 000 N N 2 2 1 0 0 Above 30 C ambient temperature this temperature may not be achieved at this speed N indicates that the rotor cannot achieve the desired temperature at this speed J Series Rotors and Tubes B 5 Temperature Compensation Tables B 6 Table B 3 Temperature Compensation Settings for the J2 MI J2 21M J2 MC and J2 21M E Centrifuges Interpolate if intermediate values are required Required Sample Temperature C green bar Speed Rotor rpm 20 C 10 C 226 EC 10 C 2020 40 C d an N N 7 5 6 5 9 N 5 5 5 5 4 7 JA 25 50 15 000 0 2 3 3 2 2 6 0 000 0 0 E 0 24 0 23 21 000 N N 8 8 7 6 9 18 000 N 8 8 8 4 5 5 denuo 15 000 6 6 B 4 4 3 2 10 000 3 2 E x 1 A 2B 21 000 N N 10 10 el 8 6 18 000 N 8 8 2 6 5 3 a 15 000 6 6 5 4 4 3 N 10 000 3 2 zl 1 0 N 20 000 N N 10 9 9 9 7 18 000 N N 7 7 7 6 4 PAPE 15 000 7 5 4 4 4 3 N 10 000 2 1 1 0 0 0 N 20 000 N N N 11 10 9 6 18 000
61. N N 8 7 5 7 6 3 ance 15 000 5 4 4 4 3 3 el 10 000 2 2 2 1 1 0 0 18 000 N N N N 8 5 0 17 000 N N N N 6 5 0 JA 18 1 15 000 N N 9 5 4 3 0 10 000 7 7 3 3 2 0 2 8 000 6 5 3 i 0 42 18 000 N N N N N 10 7 17 000 N N N 10 9 8 6 16 000 N N 10 10 9 7 5 verte 12 000 N 6 6 6 5 4 3 8 000 5 4 4 3 2 1 0 5 000 4 3 2 2 1 0 0 17 000 N N f 7 7 4 2 15 000 N 5 4 4 4 3 1 nd 12 000 4 4 3 3 3 1 N 8 000 ES 0 0 0 0 N 14 000 N N 7 5 4 3 3 JLA 16 250 10 000 N 4 3 3 2 1 N 5 000 N 0 0 0 0 0 N 14 000 N N ae e er 5 4 12 000 N 6 5 5 4 3 2 deel 10 000 4 4 3 3 2 1 N 5 000 0 0 0 0 0 N Above 30 C ambient temperature this temperature may not be achieved at this speed Continued N indicates that the rotor cannot achieve the desired temperature at this speed J Series Rotors and Tubes Temperature Compensation Tables Table B 3 Temperature Compensation Settings for the J2 MI J2 21M J2 MC and J2 21M E Centrifuges continued 12 000 N 6 6 7 7 6 9 JA 12 10 000 3 4 5 5 5 5 9 5 000 1 3 3 4 4 5 9 10 000 N 3 2 1 0 1 2 JA 10 8 000 3 2 1 1 0 1 2 5 000 2 0 0 0 0 1 2 10 000 N 5 2 1 0 42 1 JLA 10 500 8 000 1 1 0 2 2 3 1 5 000 0 1 2 1 4 3 1 13 000 N 10 10 10 10 10 9 11 000 N 8 8 8 7 6 5 Jeden 8 000 6 5 4 4 3 3 N 5 000 4 3 1 1 1 1 N 7 500 4 3 1 1 1 2 3
62. NTS AND TYPICAL USAGE SINGLE DONOR FRESH PLASMA Single donor fresh plasma is separated from whole blood within 4 to 8 hours after collection If it is not used immediately it may be frozen and stored see SHELF LIFE below Fresh and fresh frozen plasma contain all plasma clotting factors Fresh plasma used for the treatment of deficiencies of clotting factors V XL and XIII e Factor VII Concentrate separated from platelet rich plasma useful for treating clotting deficiencies other than those mentioned above e Cryoprecipitated Antihemophilic Factor factor VIIT a protein concentrate separated from cell free plasma frozen then thawed at 4 C useful for treating hemophilia Platelet Concentrates separated from plasma platelet concen trates are used to treat decreased platelet counts or failing platelet functions Platelets must be separated by centrifugation within 6 hours of collection SINGLE DONOR PLASMA Single donor plasma can be separated from whole blood up to a few days after the expiration date since no attempt is made to maintain the activity of the labile clotting factors This plasma which may be frozen and kept for up to 5 years is used for expansion of blood volume treatment of hypovolemic shock caused by a dangerous reduction in blood volume D 2 J Series Rotors and Tubes PACKED RED BLOOD CELLS RBC RBC required when the oxygen carrying capacity must be improved without overloadi
63. ORMATION ROTOR BALANCE J J Series Rotors and Tubes I Careful centrifugation technique is essential because forces gener ated in high speed centrifugation can be enormous For example 10 grams at the bottom of a JA 25 50 fixed angle rotor rotating at 25 000 rpm exerts the gravitational equivalent of 0 8 ton of centrifugal mass at the bottom of the tube cavity III NOTE Specific information about filling sealing and capping containers loading rotors etc is contained in later sections The mass of a properly loaded rotor is evenly distributed on the centrifuge drive hub causing the rotor to turn smoothly with the drive An improperly loaded rotor will be unbalanced consistent running of unbalanced rotors will reduce centrifuge drive life To balance the rotor load fill all opposing containers to the same level Rotors ROTOR TIE DOWN Knob l Tie down Assembly with liquid of the same density Weight of opposing containers must be distributed equally Place tubes in a fixed angle vertical tube or JS 24 series swinging bucket rotor symmetrically as illustrated in Figure 1 6 Detailed information about balancing other swinging bucket rotors is contained in Section 5 USING SWINGING BUCKET ROTORS If sample quantity is limited and the rotor is not balanced do one of the following to balance the rotor depending on the rotor in use Load the opposite rotor cavities or buckets with tubes c
64. Operation us oat e op ocEXO tq oce bina ped d Spade ace Installing the Rotor leseeee ee eee Removal and Sample Recovery 00 00 e ee eee USING SWINGING BUCKET ROTORS Description 2 os nei esas ee eee hea ee ee RI RE d ES Labware vectes eese da eth oe ab tal eins Rotor Preparation and Loading 0 0 0 e eee ee eee Prerun Safety Checks 0 0 cece eee eee ee Rotor Preparation csse er ee ba ae Loading the Rotor Yoke 0 0 0 cece eee ee eee Symmetric and Balanced Loading Loading Buckets c 05 acce Rr Ree ne de Loading Buckets into the Rotor 000 Using Microtiter Plate Carriers 0 000000 eee Operation els repere prece Bee b eof ees sample Recovety uda sase tue ER de donare eb Ee bs USING VERTICAL TUBE AND RACK TYPE ROTORS Description 2 3 66 esas d de l RR ERR UY pede ee det hg Vertical Tube Rotors 0 0 eee eee Rack Type Rotors iu eio ek PER D ua RHENO ete Using a Vertical Tube Rotor 0 0 0 0 cece eee ee Tubes and Bottles 1 2 cca eere e eee aly Rotor Preparation and Loading 0 0 00 0000 Operation i bese ee ee ee be Rr ERE ee ee d RR Installing the Rotor 0 2 eee eee eee Removal and Sample Recovery 0 000 Page Contents Contents Page Using a Rack Type Rotor 0 0 0 cee eee eee eee 6 7 Trays and Tubes used teenime CET ERE EAE eue 6 7 R
65. ROSOLVE M CANNISTERS Aerosolve Cannister Aerosolve Tube Rack BLOOD BAG CUPS 2 12 Aerosolve cannisters used in the JS 4 3 swinging bucket rotor are designed to minimize aerosol leakage and liquid spills The cannister is transparent enabling you to see broken labware and take proper precautions before opening the cannister The cannister and lid are made of polyphenylsulfone tube racks are made of polypropylene and the O ring is ethylene propylene rubber Refer to Appendix A Chemical Resistances to determine compatibil ities with specific chemicals Each cannister can hold a variety of tube sizes in tube racks that are specifically designed to fit in the cannisters The cannister can also be used as a 500 mL wide mouth bottle N WARNING When centrifuging hazardous materials always open cannisters in an appropriate hood or biological safety cabinet Polypropylene blood bag cups are available for use in swinging bucket rotors to obtain cell free plasma for cell packing or for leukolyte depletion Different sizes of cups are available to accommo date single double triple or quad pack blood bags Refer to the applicable rotor manual to determine the correct blood bag cup to use Blood bag cups are autoclavable Blood bags should be loaded into the cups outside of the centrifuge to avoid tripping the centrifuge imbalance monitor during loading J Series Rotors and Tubes Tubes Bottles and Accessories
66. Temperature Compensation Settings for the J2 HC Centrifuge continued N 12 000 N 8 3 2 1 JAIA 10 000 4 E N 5 000 1 0 N 12 000 N 2 1 JA 12 10 000 1 1 5 000 0 0 10 000 1 2 JA 10 8 000 1 2 5 000 1 2 10 000 E 0 JLA 10 500 8 000 1 0 5 000 0 2 13 000 6 8 11 000 4 e EPA 8 000 1 N 5 000 0 N 7 500 4 4 JS 7 5 5 000 3 3 2 000 2 N 4 300 0 2 JS 4 3 3 000 0 2 1 500 0 N 18 000 6 6 JV 20 15 000 3 4 10 000 1 0 N CAUTION For proper temperature control the JA 18 1 fixed angle rotor must be derated in the J2 HC Centrifuge as follows when the 25 degree angle adapters are used the maximum speed is 16 000 rpm when the 45 degree angle adapters are used the maximum speed is 15 000 rpm Above 30 C ambient temperature this temperature may not be achieved at this speed N indicates that the rotor cannot achieve the desired temperature at this speed J Series Rotors and Tubes B 3 Temperature Compensation Tables B 4 Table B 2 Temperature Compensation Settings for the J2 21 J2 21B J2 21C and J2 HS Centrifuges Interpolate if intermediate values are required 20 000 N N 7 zB 6 M 9 18 000 N B ig 5 5 4 E UAREDSU 15 000 0 2 3 8 2 2 6 10 000 0 0 4 0 E 0 8 21000 N N 10 10 7 6 9 18 000 N 8 8 E 4 5 5 IS 15 000 6 6 5 4 4 3 2 10 000 3 2 4 E 4 4 2 21000 N N 10 10 10 8 6 18 000 N 8
67. Y TO THE FACTORY Before returning a rotor or accessory for any reason prior permission a Returned Goods Authorization form must be obtained from Beckman Coulter Inc This RGA form may be obtained from your local sales office It should contain the following information serial number history of use approximate frequency of use reason for the return original purchase order number billing number and shipping number if possible name and phone number of the person to be notified upon receipt of the rotor or accessory at the factory and name and phone number of the person to be notified about repair costs etc To protect our personnel it is the customer s responsibility to ensure that the parts are free from pathogens chemical hazards and or radioactivity Sterilization and decontamination MUST be done before returning the parts Smaller items such as tubes bottles and so on should be enclosed in a sealed plastic bag All parts must be accompanied by a note plainly visible on the outside of the box or bag stating that they are safe to handle and that they are not contaminated with pathogens chemical hazards or radioactivity Failure to attach this notification will result in return or disposal of the items without review of the reported problem Use the address label printed on the RGA form when mailing the rotor and or accessories
68. again will remove the minus sign Check the temperature display If the entry is incorrect press and reenter the digits When the entry is correct press ENTER RECALL J Series Rotors and Tubes 2 Tubes Bottles and Accessories This section describes various labware used in Beckman Coulter J series rotors General instructions for using containers follow in Section 3 Care and maintenance instructions are in Section 7 General rotor use instructions are in Sections 4 through 6 The indi vidual rotor manual that comes with each rotor provides specific instructions on the tubes bottles and accessories that can be used in a particular rotor A table of chemical resistances can be found in Appendix A of this manual LABWARE SELECTION CRITERIA No single tube or bottle design or material meets all application requirements Labware choice is usually based on a number of factors e The centrifugation technique to be used including the rotor in use quantity of sample to be centrifuged need for sterilization importance of band visibility and so forth e Chemical resistance the nature of the sample and any solvent or gradient media Temperature and speed considerations Whether tubes or bottles are to be reused Table 2 1 contains an overview of some of the characteristics of tube and bottle materials A complete list of tubes bottles and accessories is provided in the latest edition of the Beckman Coult
69. ailure during subsequent runs JS 5 0 CUPS 1 Remove the rotor lid and hang it on the black rubber block on the inside of the centrifuge door 2 Remove the plug from the labware cup cover and pour the super natant out of the cup through the spout Or remove the cup cover and pour the supernatant over the cup edge 3 Ifaliner was used remove the liner from the cup Fold or heat seal the liner5 and store the pellet as required 5 Beckman Coulter recommends Cole Parmer heat sealer Model U 03018 10 adjusted to setting 3 or 4 Contact Cole Parmer at 800 323 4340 by Fax at 847 247 2929 or at www coleparmer com 3 20 J Series Rotors and Tubes Using Tubes Bottles and Accessories 4 Ifa liner was not used first remove the red cup gasket 369257 remove the partition if used and then use the spatula 367891 ae LLL p a to remove pellet from the cup Do not use a metal tool to remove pellet as metal could damage the cup and shorten its useful life QUICK SEAL TUBES There are several methods of recovering fractions from Quick Seal tubes One of the following procedures may be used gt NOTE iS Kei eA E If you plan to collect particles from the tube side provide an or bottom first create an air passage by snipping aris the stem or inserting a hollow hypodermic needle in the top of the tube Puncture the side of the tube just below the band with a needle and syringe and draw the sample off Take care when
70. and Tubes 5 3 Using Swinging Bucket Rotors Table 5 1 General Specifications for Beckman Coulter J Series Swinging Bucket Rotors continued JS 4 3 4300 rpm 400 204 buckets 4x 750 mL 3 liters 4220 x g to 163 carriers 4 blood bags 16 635 1450 12 microplates 148 RIA tubes JS 4 2 4200 rpm 600 254 184 114 6 x 1 liter 6 liters 5020 x g to 6 blood bags 11502 800 18 microplates 336 RIA tubes JS 4 2SM 4200 rpm 600 248 182 116 6 triple or 6 liters r 3 4900 x g to quad pack 6 800 blood bags JS 4 2A 4200 rpm 600 254 184 114 6 x 1 liter 6 liters use only in 5020 x g to 6 blood bags J6 series 11 502 800 18 microplates centrifuges 336 RIA tubes JS 4 2SMA 4200 rom 600 248 182 116 6 triple or quad 6 liters use only in 4900 x g to pack blood bags J6 series 800 centrifuges JS 4 0 4000 rpm 600 226 156 86 4 x 1 liter 4 liters 4044 x g to 4 blood bags 15 296 800 12 microplates 148 RIA tubes JS 3 0 3000 rpm 600 254 184 114 6 x 1 liter 6 liters 2560 x g to 6 blood bags 22 548 800 18 microplates 336 RIA tubes JS 2 9 2900 rpm 600 265 192 118 12 x 500 mL 6 liters 2500 x g to blood bags 24 400 800 Maximum speeds are based on a solution density of 1 2 g mL Critical speed range is the range of speeds over which the rotor shifts so as to rotate about its center of mass Passing through or running at the critical speed range is characterized by
71. and sample IB NOTE Partially filled tube racks should contain the same number of balanced tubes Each tube in a rack must be balanced by a tube in a diametri cally opposed position in the opposite rack Using Tubes Bottles and Accessories Table 3 2 Aerosolve Tube Racks white 1 5 11 24 354495 none blue 3 amp 5 12 24 359160 359482 344117 tan 5 13 24 358993 359489 none orange 10 14 18 359161 359483 344118 purple 12 16 12 359162 359484 344119 3 amp 5 12 6 white 15 14 10 344517 none vials green 15 amp 20 18 12 359163 359485 344120 3 amp 5 12 6 light 15 T 6 358991 359487 none green 3 amp 5 12 6 conical lime green 50 30 4 358992 359488 none conical 3 amp 5 12 4 yellow 50 29 4 359164 359486 344121 3 amp 5 12 4 These cushions are inserted into the tube holes in the base of the rack An additional pad part number 349948 inserted in the rotor bucket beneath the entire cannister is also needed If using 15 mL Vacutainers only four may be loaded into this tube rack the two outer positions are restricted by the cover height Vacutainer is a registered trademark of Beckton Dickinson and Company USING BLOOD BAG CUPS N WARNING Ask your laboratory safety officer to advise you about the level of containment required for your application and about the proper decontamination or sterilization procedures to follow if fluids escape from containers
72. ated using a solution that will not damage the anodized surfaces Beckman Coulter has tested a number of solutions and found two that do not harm anodized aluminum RadCon Surface Spray or IsoClean Solution for soaking and Radiacwash I gt NOTE IsoClean can cause fading of colored anodized surfaces Use it only when necessary and do not soak rotor components longer than the minimum time specified in the IsoClean usage instruc tions Then remove it promptly from surfaces 1 In U S contact Nuclear Associates New York in Eastern Europe and Commonwealth States contact Victoreen GmbH Munich in South Pacific contact Gammasonics Pty Ltd Australia in Japan contact Toyo Medic Co Ltd Tokyo 2 n U S contact Biodex Medical Systems Shirley New York internationally contact the U S office to find the dealer closest to you J Series Rotors and Tubes ges Care and Maintenance While Beckman Coulter has tested these methods and found that they do not damage components no guarantee of decontamination is expressed or implied Consult your laboratory safety officer regarding the proper decontamination methods to use Ifthe rotor or other components are contaminated with toxic or pathogenic materials follow appropriate decontamination proce dures as outlined by appropriate laboratory safety guidelines and or other regulations Consult Appendix A to select an agent that will not damage the rotor STERILIZATION AND
73. atures between 54 and 121 C 65 and 250 F E 3 Glossary of Terms Noryl pelleting PET plasma plasmapheresis plateletpheresis platelets polyallomer rack type rotor Radel rate zonal RBC modified polyphenylene oxide used in some bottle caps Noryl is a registered trademark of GE Plastics a centrifugal separation process in which particles in a sample sediment to the bottom of the tube differential separation differential pelleting separates particles of different sizes by successive centrifugation steps of progressively higher g force and or longer run duration polyethylene terephthalate used in some adapters major component of blood made up primarily of water with substances such as albumin globulins coagulation factors and electrolytes distributes nutrients to the body absorbs and carries away waste products procedure in which whole blood is collected platelets are separated via centrifugation and platelet poor red blood cells are returned to the donor plasma is returned to the donor or collected for fractionation into clotting factors and albumin procedure in which a unit of blood is taken to obtain plasma following blood separation red cells are immediately reinfused to the donor blood component responsible for blood coagulation random block copolymer of ethylene and propylene used for certain tubes Tenite Polyallomer is a registered trademark of Eastman Chemical Co aro
74. bes with the Beckman Coulter Cordless Tube Topper M Use and Care of Centrifuge Tubes and Bottles Documents referenced below are available upon request from Beckman Coulter Inc Marketing Communications 4300 N Harbor Blvd Box 3100 Fullerton CA 92834 U S A Rapid Pelleting of Bacteria in the Avanti J Centrifuge High Performance High Speed High Capacity Rotors Tubes amp Accessories Use of the w t Integrator Density Gradient Separations in Vertical Tube Fixed Angle and SW Rotors Bibliography of Applications of the Vertical Tube Rotor Use of k Factor for Estimating Run Times F 1 References DS 728 Optimizing Centrifugal Separations Sample Loading DS 746 Beckman Coulter Tubes and Bottles DS 776 Using k Factor to Compare Efficiency of Fixed Angle Rotors DS 797 New Large Capacity Multitube Carrier Holds 12 x 75 mm Tubes in the JS 7 5 Rotor DS 829 Method for Plasmid DNA Mini Preparation Using the JA 18 1 Rotor DS 885 Using the JA 18 Rotor to Process Large Volumes Rapidly DS 910 New JA 12 Rotor for High Speed Processing of 50 mL Conical Tubes Without Adapters DS 8028 The JLA 10 500 Rotor with 3 Liter Capacity DS 9240 AllSpin JS 5 3 Rotor for Avanti J E and J 20XP centrifuges SB 812 J6 Series High Capacity Centrifuges T 1735 Modification of the Model J6 MC for Blood Component Preparation T 1741 Optimizing Radioimmunoassays with the JR 3 2 Rack Rotor F 2 J Series Rotors and Tubes SERIES ROTOR WARRANTY
75. cket If at the time of purchase the marked expiration date is less than 7 years from the date of purchase the expiration date becomes the date of purchase plus seven 7 years Use of a bucket after such expiration date voids Beckman Coulter s warranty obligations with respect to any rotor and or centrifuge in which such a bucket is used DISCLAIMER IT IS EXPRESSLY AGREED THAT THE ABOVE WAR RANTY SHALL BE IN LIEU OF ALL WARRANTIES OF FITNESS AND OF THE WARRANTY OF MERCHANT ABILITY AND THAT BECKMAN COULTER INC SHALL HAVE NO LIABILITY FOR SPECIAL OR CON SEQUENTIAL DAMAGES OF ANY KIND WHATSO EVER ARISING OUT OF THE MANUFACTURE USE SALE HANDLING REPAIR MAINTENANCE OR REPLACEMENT OF THE PRODUCT Beckman Coulter Worldwide Life Science Research Division Offices AUSTRALIA Beckman Coulter Australia Pty Ltd Unit D 24 College St Gladesville NSW 2111 Australia Telephone 61 2 9844 6000 or toll free 1 800 060 880 Fax 61 2 9844 6096 email lifescienceaustralia beckman com CANADA Beckman Coulter Canada Inc 6755 Mississauga Road Suite 600 Mississauga Ontario Canada L5N 7Y2 Telephone 905 819 1234 Fax 905 819 1485 CHINA Beckman Coulter Inc Beijing Representative Office Unit 2005A 2006 2009 East Ocean Center Jian Guomenwai Avenue Beijing 100004 China Telephone 86 10 6515 6028 Fax 86 10 6515 6025 6515 6026 EASTERN EUROPE MIDDLE EAST NORTH AFRICA Beckman Coulter Internatio
76. commonly used gradient materials Appendix D provides information about separation of blood components using J series centrifuges Appendix E provides a glossary of terms Appendix F lists references for further reading J Series Rotors and Tubes J Series Rotors and Tubes Contents Page Scope of this Manual SECTION 1 ROTORS General Description o csacsi 0 0 cece eee eee eens 1 1 Rotor Designations 0 0 cece eee eee 1 1 Materials 42 evene RE EP Roe edna aay 1 3 Drive PINs obese bLRCE ee E HER EE a da 1 4 Rotor Selection 2 cse aden eR ida eae aH rer bbe 1 5 Pelleting Differential Separation 4 1 10 Isopycnic Separations 0 0 eee eee eee eee 1 13 Rate Zonal Separations 0 0 cece eee eee eee 1 14 Blood Component Separations 0 000 000 008 1 15 General Operating Information 0 000020 e ee 1 15 Rotor Balance i eiae ee e aem boa weg 1 15 Rotor Tie down lesse t mme ee 1 16 Overspeed Protection 0 0 eee eee eee 1 17 Allowable Run Speeds 0 0 00 ce eee eee eee 1 18 Temperature Compensation 0 0 0 0 eee ee eee 1 19 SECTION 2 TUBES BOTTLES AND ACCESSORIES Labware Selection Criteria 2 1 0 eee ee ee eee 2 1 Labware Material Compatibility with Solvents and Sample 2 3 Gradient Formation and Fractionation 2 3 Labware Types eec rene eere e a ce qe 2 3 Polyallomer Tubes
77. cover wrong type of bucket mixed bucket types opposing buckets not filled to the same level with liquids of the same density Check loading procedures refer to Section 5 e Swinging bucket rotor Pivot pins and bucket pin pockets not lubri cated every 250 runs Lubricate as described in LUBRICATION earlier in this section Critical speed range is the range of speeds over which the rotor shifts so as to rotate about its center of mass Passing through the critical speed range is characterized by some vibration 7 12 J Series Rotors and Tubes Care and Maintenance SYMPTOM POSSIBLE CAUSE AND SUGGESTED ACTION Rotor lid cannister cover or bucket Vacuum built up inside the container during centrifugation Lift the cover is difficult to remove after vent plug on bucket or cannister covers with vents to relieve the centrifugation vacuum Threads contaminated with dirt dried lubricant or metal particles or threads insufficiently lubricated cause rotor components to stick Do not use excessive force to loosen components Contact your Beckman Coulter representative Routinely clean metal threads with concentrated Solution 555 then lubricate them with Spinkote Anodizing coming off where bucket Not an operational problem some buckets are not anodized inside the pin or carrier contacts rotor pins on pockets to facilitate swinging swinging bucket rotor Adapters stick in buckets after Apply a thin film of powder such as
78. ction manual for detailed operating information For low temperature runs precool the rotor in the centri fuge or in a refrigerator before use especially before short runs to ensure that the rotor reaches the set temperature To ensure that the rotor reaches the required temperature during centrifugation some temperature compensation may be required because of the mass of these rotors Refer to Appendix B or to the rotor manual for tables listing temperature compensation units for various rotors If you are using a microprocessor controlled J2 or J6 series centri fuge enter the rotor code if the JS 5 2 rotor is used for example enter code 5 2 J Series Rotors and Tubes Using Swinging Bucket Rotors gt NOTE These rotors are not used in Avanti J series centrifuges except the Avanti J HC SAMPLE RECOVERY N CAUTION If disassembly reveals evidence of leakage you should assume that some fluid escaped the container or rotor Use appropriate decontami nation procedures on the centrifuge rotor and accessories 1 Remove the rotor lid if applicable Remove the buckets or carriers from the rotor 2 Remove labware from the buckets or carriers I gt NOTE Except for the JS 24 38 and JS 24 15 rotors you can leave the rotor body or yoke in the centrifuge between runs unless spillage has occurred in which case you should remove the buckets or carriers and yoke and clean the centrifuge and rotor compon
79. d sets will help you keep them together It is not necessary to completely fill all tubes positions in adapters or wells in microtiter plates however partially filled adapters or micro titer plates must be balanced with respect to the pivotal axis of the bucket or carrier as discussed below During a run buckets and carriers swing 90 degrees from their at rest position The pivotal axis of a bucket or carrier can be imagined as a line extending across the bucket or carrier from one pivot pin to the other If a bucket or carrier is loaded so that its weight is unequally distributed on either side of its pivotal axis it will not hang vertically at rest and more importantly may not swing to a horizontal position during arun As a result extra stress will be placed on the bucket catrier tubes and or microtiter plates during the run increasing the possibility of breakage or rotor imbalance J Series Rotors and Tubes Using Swinging Bucket Rotors LOADING BUCKETS Buckets can be loaded before or after being installed on the rotor yoke For best results fill the appropriate labware first and then load the labware into the buckets This is especially important when using blood bags you can trip the imbalance detector in the centrifuge by pushing blood bags into cups within buckets that are installed in the rotor You can also bend the centrifuge drive spindle Using Modular Disk Adapters Assemble modular disk adapters as follows
80. de of Hytrel3 and do not require coating Currently produced J series rotors have drive pins in the drive hole These pins mesh with teeth on the centrifuge drive spindle hub when the rotor is installed to ensure that the rotor does not slip on the hub during initial acceleration Most drive pins are oriented horizontally or angled in the drive hole however some are oriented vertically All rotors used in Avanti J series centrifuges must have drive pins in the rotor drive hole Some Beckman Coulter rotors including the JA 10 and the JS 7 5 were previously manufactured without drive pins because pins were not needed when these rotors were used in J2 series centrifuges Check all J series rotors for drive pins before using them in an Avanti J series centrifuge To check for drive pins hold the rotor up or turn it on its side and look into the drive hole If you do not see two metal pins near the top of the hole do not use the rotor in the Avanti J Call your local Beckman Coulter office for information on returning the rotor to the factory for upgrading In fixed angle and vertical tube rotors manufactured since early 1997 the rotor pins are positioned parallel to the BECKMAN name engraved at the center of the rotor body Knowing the pin orientation before you install the rotor will help to ensure that you position the rotor properly on the hub minimizing the chance of hub damage 3 Hytrel is a registered trademark of E I Du Pont de Nemo
81. e imbalance In addition the J6 adapter bails will interfere with the JS 4 3 rotor yoke when the buckets swing up to horizontal position BOTTLE AND TUBE CAPS The need for caps depends on such factors as the kind of rotor being used the type of container and the amount of sample being centrifuged Some tubes must be capped before centrifugation as in the case of thinwall tubes The thickness and strength of some containers such as thickwall plastic and stainless steel tubes used in fixed angle and swinging bucket rotors allows them to be run without caps but they must be only partially filled Refer to the applicable rotor manual for allowable capless fill levels When greater fill volumes are required in these tubes caps must be used for sample retention When closed containers are required several choices are available e Cap assemblies threaded caps with inserts and O rings or one piece caps with O rings that provide a leakproof closure to accommodate a capacity container load that is to the bottom of the insert Threaded caps without inserts or O rings these are not as liquid tight as the cap assemblies therefore the meniscus must be kept lower to prevent leakage e Snap on caps these caps are simple to use but are not as liquid tight as the cap assemblies or threaded caps They require an even lower meniscus to prevent leakage J Series Rotors and Tubes 2 11 Tubes Bottles and Accessories AE
82. e applicable rotor manual that corresponds with the required temperature and run speed Set the COMP dial to that setting Interpolate if inter mediate values are required I gt NOTE Temperature settings for J 21 series centrifuges must be empirically determined Rotors Analog J6 Series Centrifuges Set temperature compensation in analog J6 model centrifuges models J6 HC and J6 B as follows 1 Find the compensation value in Appendix B or in the applicable rotor manual that corresponds with the required temperature and run speed Interpolate if intermediate values are required Turn the SET knob on the centrifuge control panel to the required sample temperature Microprocessor Controlled Centrifuges Operating temperatures for most rotors are contained in memory in microprocessor controlled centrifuges models J2 MI J2 21M TEMPERATURE C J2 MC and J2 21M E Set temperature compensation as follows for rotors not in centrifuge memory 1 20 1 Press the key on the centrifuge control panel and then use the keypad to enter the required sample temperature Find the compensation value in Appendix B or in the applicable rotor manual that corresponds with the required temperature and run speed Press COMP ADJ The word COMP flashes in the TEMPERATURE display and the display flashes Use the keypad to enter the compensation value Press the key to enter a minus sign pressing it
83. e cavity are the same Using adapters more than one type and size of tube can be centrifuged together provided that the load 1s properly balanced O rings made of Buna N rubber are located in the rotor lid The O rings help to maintain atmospheric pressure inside a fixed angle rotor during centrifugation when they are properly lubricated A tie down device or lid locking knob is used to secure the rotor to the centrifuge drive spindle hub before the run begins Using Fixed Angle Rotors 34 f ri ri rd r min lav T max Axis of Rotation r min As g V A T max Axis of Rotation Axis of Rotation JA 20 JA 25 15 JLA 10 500 Figure 4 1 Examples of Fixed Angle Rotors J Series Rotors and Tubes Using Fixed Angle Rotors Table 4 1 General Specifications for Beckman Coulter J Series Fixed Angle Rotors JA 30 50 Ti 30 000 rpm 600 to 108 74 40 8x50mL 400 mL 34 Angle 108 860 x g 800 280 JA 25 50 25 000 rome 600 to 108 73 2 38 5 8 x 50 mL 400 mL 34 Angle 75 600 x g 800 418 JA 25 15 25 000 rpm 600 to 24x15mL 360 mL 25 Angle 74200xg 800 106 79 52 outer row 265 60200xg 86 59 32 inner row 380 JA 21 21 000 rpm 600 to 102 73 45 18 x 10 mL 180 mL 40 Angle 50400xg 800 470 JA 20 1 20 000 rpm
84. e high speed centrifuges can achieve temperatures as high as 45 C only certain tube or bottle materials can be run under these conditions Most containers are made of thermoplastic materials that soften at elevated temperatures This temperature induced softening together with such factors as the centrifugal force the run duration the type of rotor previous run history and the tube angle can cause labware to collapse Therefore if high temperature runs above 25 C are required it is best to pretest labware under the actual experimental conditions using buffer or gradient of similar density rather than a valuable sample Stainless steel tubes can be centri fuged at any temperature 25 C ea D e Plastic labware has been centrifuge tested for use at temperatures between 2 and 25 C For centrifugation at other temperatures pretest tubes under anticipated run conditions e f plastic containers are frozen before use make sure that they are thawed to at least 2 C prior to centrifugation 2 8 J Series Rotors and Tubes Tubes Bottles and Accessories SPACERS AND FLOATING SPACERS Quick Seal tubes require spacers made of anodized aluminum with Spacer or without floating spacers The particular combination depends on the type of rotor being used and the tube size Floating Spacer e In swinging bucket and fixed angle rotors the top of the tube must be supported n vertical tube rotors the entire cavity must be
85. e is the range of speeds over which the rotor shifts so as to rotate about its center of mass Passing through or running at the critical speed range is characterized by some vibration Maximum speed in an Avanti J E centrifuge is 21 000 rpm 4 When a JA 18 1 rotor is used in the J2 HC centrifuge derate the rotor as follows when the 45 adapters are used do not run the rotor above 15 000 rpm when 25 adapters are used do not run the rotor above 16 000 rpm Maximum speed in an Avanti J series centrifuge except Avanti J E maximum speed in an Avanti J E for a rotor with magnets maximum speed for rotor without magnets is 13 000 rpm Maximum speed in a J2 series centrifuge is 14 000 rpm f Maximum speed in an Avanti J E for a rotor with magnets maximum speed for a rotor without magnets is 13 000 rpm Maximum speed in an Avanti J E for the rotor with magnets without magnets maximum is 14 000 rpm Maximum speed at 2 C in a 50 Hz centrifuge is 14 000 rpm Do not put bottles directly into the rotor without cannisters J Series Rotors and Tubes 4 5 Using Fixed Angle Rotors Rotor Knob O Ring Sealing Surface Fluid Containment Annulus TUBES AND BOTTLES Some rotors have dual locking lid mechanisms consisting of a daisy knob that secures the lid to the rotor and a round rotor knob that attaches the rotor to the centrifuge drive spindle hub Daisy refers to the knob shape The grooves between each petal let y
86. e of rotor components Many rotors use O rings or gaskets as seals to maintain atmospheric pressure in the rotor during a run These O rings and the rotor surfaces they bear against must be kept clean and evenly lubricated 7 5 Care and Maintenance Pivot Pins and Buckets Lubrication Points between the pivot pin and rotor yoke all pivot pins Rotor Yoke y Lubrication Points all pin surfaces and both pin sockets After removing and cleaning rotor or bucket O rings or gaskets lightly but uniformly coat them with silicone vacuum grease and reposition them in the rotor I gt NOTE Do not apply lubricant with a cotton tipped swab These swabs can leave lint on the O ring or gasket that can interfere with the seal JS 4 2 JS 4 2A and JS 4 2SMA Rotors Approximately every three months or every 250 runs and after cleaning and or autoclaving the rotor lubricate the contact areas between the buckets and the pivot pins 1 Saturate a lintless tissue with Tri Flow oil 883371 2 Wipe the oil soaked tissue on each bucket socket and pivot pin 3 Put an additional drop of Tri Flow between each pivot pin and the yoke JS 5 9 JS 5 3 JS 5 0 and JS 4 3 Rotors Approximately once a week and after cleaning and or autoclaving lubricate the pin sockets with a lubricant such as Paint On Graphite Lubricant 977212 Allow the lubricant to dry for at least 5 minutes before installing the rotor in a centrifuge
87. e reduced rotor speed for optimum labware performance Refer to the applicable rotor manual for bottle fill levels and cap requirements 3 7 Using Tubes Bottles and Accessories THREE PIECE CAP ASSEMBLIES Cap Cm Cap bottles with three piece cap assemblies as follows iu 1 Be sure the O ring plug and bottle lip are dry and free of ang Bottle lubrication Lip 2 Place the O ring on the underside of the plug 3 Insert the plug into the neck of the bottle ensuring that no fluid contacts the O ring 4 Tighten the cap by hand JLA 8 1000 AND JLA 9 1000 BOTTLE CAP CLOSURES Tighten the cap closure unt Place the plug on the bottle then screw on the cap closure by hand as this mark aligns with or goes tightly as possible Tighten until the timing mark on the cap closure is past ING DONE VERNE aligned with or goes past the vent line on the bottle MAX FILL LINE l 1000mL Vent Line FILLING AND LOADING CUPS IN THE JS 5 0 ROTOR I gt NOTE Four labware cups must be used for every run and must be balanced to within 25 grams of each other Do not load the rotor with two filled cups and two empty cups 1 Insert four labware cups into two cup racks with the cup latch hinges toward the center of the racks 2 Make sure that the gaskets and sealing surfaces on each cup and cover are clean and dry Place a gasket around the top edge of each cup carefully pushing the gasket down until it is fu
88. el Centrifuges Using Fixed Angle Rotors In older model centrifuges be sure the pins in the rotor drive hole are located in the grooves of the drive spindle hub nnewer model centrifuges be sure the pins in the rotor drive hole are not sitting on top of the teeth on the drive spindle hub N CAUTION The pins located in the rotor drive hole must be seated correctly on the centrifuge drive spindle Running a rotor that is not seated properly may result in severe rotor damage 2 After the rotor is seated on the drive spindle hub place the lid on the rotor 3 Press down on the knob then screw it down tight Turning the knob to the right clockwise attaches the rotor to the hub the lid on some fixed angle rotors remains free and may be slipped on or off while the rotor remains secured in the centrifuge IB NOTE The JA 18 rotor must be run with the lid on in Avanti J series centrifuges REMOVAL AND SAMPLE RECOVERY J Series Rotors and Tubes AN CAUTION If disassembly reveals evidence of leakage you should assume that some fluid escaped the container or rotor Apply appropriate decontam ination procedures to the centrifuge rotor and accessories 4 9 Using Fixed Angle Rotors Lifting Handle Assembly 346965 Quick Seal Tube Removal Tool 361668 4 10 Unscrew the rotor lid knob to release the rotor from the spindle hub gt NOTE Labware can
89. ent potential rotor failures Contact your local Beckman Coulter office to request this service It is your responsibility to decontaminate the rotors and accessories before requesting service by a Beckman Coulter Field Service representative J Series Rotors and Tubes JR IM 6 August 2002 COULTER ROTORS AND TUBES For Beckman Coulter J2 J6 and Avanti J Series Centrifuges User s Manual 2002 Beckman Coulter Inc Printed in the U S A Published by the Centrifuge Instrument Systems Development Center Of Beckman Coulter Inc Palo Alto CaliforniA 94304 SCOPE OF THIS MANUAL This manual contains general information for properly preparing a rotor for centrifugation in a Beckman Coulter J series centrifuge This manual should be used with the individual rotor instruction manual packed with each rotor The rotor manuals provide specific information for each rotor including special operating procedures and precautions tube bottle and adapter part numbers and equations to calculate maximum allowable rotor speeds Each manual has a code number in the upper right hand corner of the cover page that can be used for reordering send your request include the code number to Technical Publications Department Beckman Coulter Inc 1050 Page Mill Road Palo Alto CA 94304 U S A Telephone 650 859 1753 Fax 650 859 1375 A lot of information is compiled in this manual and we urge you to read it care
90. ents immediately according to the instructions in the centrifuge and rotor instruction manuals If the rotor is left in the centrifuge between runs tighten the tie down device before each run The JS 24 38 and JS 24 15 rotors must be removed from the centrifuge to install or remove buckets J Series Rotors and Tubes 5 17 Using Swinging Bucket Rotors Removing JS 24 Series Rotors 1 Remove the rotor from the centrifuge by lifting it straight up and off the drive hub 2 Set the rotor on the rotor stand and carefully remove the buckets 3 Remove the bucket caps and use the appropriate removal tool listed in the rotor manual to remove the spacers and tubes If floating spacers were used remove them with the threaded end While pressing the rubber tip of the floating spacer removal tool 338765 against the adapter wall Extractor pull the tool Lone and adapter up and out Hl NOTE of the cavity If the conical shaped adapters that support konical tubes are difficult to remove after centrifugation an extractor tool 354468 is available to facilitate removal See Section 7 for instructions on the care of rotors tubes or bottles and accessories after a run 5 18 J Series Rotors and Tubes J N DESCRIPTION VERTICAL TUBE ROTORS J Series Rotors and Tubes 6 Using Vertical Tube and Rack Type Rotors This section contains instructions for using vertical tube or rack type rotors in J series centri
91. epresentative The replacement price cost to Buyer for the respective rotor shall be calculated as follows years Replacement price Current rotor list price x CONDITIONS Except as otherwise specifically provided herein this war ranty covers the rotor only and Beckman Coulter Inc shall not be liable for damage to accessories or ancillary supplies including but not limited to i tubes ii tube caps iii tube adapters or iv tube contents 2 This warranty is void if the rotor has been subjected to customer misuse such as operation or maintenance con trary to the instructions in the Beckman Coulter rotor or centrifuge manual 3 This warranty is void if the rotor is operated with a rotor drive unit or in a centrifuge unmatched to the rotor charac teristics or is operated in a Beckman Coulter centrifuge that has been improperly disassembled repaired or modified 4 Each bucket whether purchased with a rotor assembly or purchased separately is covered by this warranty for seven 7 years from the date of purchase and will be replaced or repaired during such period according to the terms and conditions of this warranty The date of manufacture marked on the bucket may be earlier than the date of pur chase and the expiration date marked on the bucket which is seven 7 years after the date of purchase may be correspondingly offset 5 Buckets should not be used after the expiration date marked on the bu
92. er High Performance High Speed High Capacity Rotors Tubes amp Accessories catalog BR 8102 available at www beckmancoulter com J Series Rotors and Tubes P Tubes Bottles and Accessories Table 2 1 Characteristics and Chemical Resistances of Tube and Bottle Materials Refer to Appendix A for information about specific solutions thinwall transparent yes yes no S U U M S U U U U U S polyallomer thickwall translucent no no yes S S S M S M M U M U S polyallomer Ultra Clear transparent yes yes no S U U S U U U U U polycarbonate transparent no no yes M U U M U U U U U M polypropylene translucent no no yes S S S M S M S M M transparent polyethylene transparent yes no yes S S S S S S U M M M S translucent cellulose transparent no no no S U U U U M S S U M S propionate stainless steel opaque no no yes S U S S M S S S M S M S satisfactory resistance M z marginal resistance U unsatisfactory resistance Polyallomer polypropylene and cellulose propionate tubes with diameters of 5 to 13 mm may be sliced using the Centritube Slicer part number 347960 and appropriate adapter plate I gt NOTE This information has been consolidated from a number of sources and is provided only as a guide to the selection of tube or bottle materials Soak tests at 1 g at 20 C established the data for most of the materials reactions may vary under the
93. er from the bucket difficult lII NOTE Do not run chloroformed samples in Aerosolve cannisters Chloroform vapors can damage the cannister material 3 Fill the cannister as described under USING CANNISTERS AS WIDE MOUTH BOTTLES or USING CANNISTERS WITH TUBE RACKS below 4 Screw the lid on until closing resistance is first felt then tighten it an additional 60 degrees The scribe marks around the rim of the cannister and the corrugated finger grips on the lid are all placed 60 degrees apart J Series Rotors and Tubes Using Tubes Bottles and Accessories USING CANNISTERS AS WIDE MOUTH BOTTLES 1 Fill each cannister only to the fill level line maximum is 500 mL of 1 2 g mL liquid 2 Run another cannister filled to the same level with liquid of the same density in the opposite bucket USING CANNISTERS WITH TUBE RACKS Aerosolve Tube Rack J Series Rotors and Tubes The racks designed to hold tubes in the Aerosolve cannister are listed in Table 3 2 Tube racks are easily disassembled by unscrewing the handle and lifting off the top plate 1 Press a rubber cushion if applicable see Table 3 2 into each tube hole in the rack base 2 Load filled tubes symmetrically into tube racks Opposing loads should weigh about the same within 10 grams Do not exceed the rated maximum load for each bucket 1000 grams Maximum bucket load includes the bucket cushion if applicable cannister tube rack tubes
94. er infec tious samples according to good laboratory procedures and methods to prevent spread of disease Because spills may generate aerosols observe proper safety precautions for aerosol containment Do not run toxic pathogenic or radioactive materials in this rotor without taking appropriate safety precautions Biosafe containment should be used when Risk Group II materials as identified in the World Health Organization Laboratory Biosafety Manual are handled materials of a higher group require more than one level of protection When working with potentially hazardous materials always fill or open containers in an appropriate hood or biological safety cabinet Three levels of containment are offered by Beckman Coulter and may be used singly or combined depending upon your application 1 Capped tubes or bottles are designed to provide fluid containment We strongly recommend that all containers carrying physiological fluids be capped to prevent leakage 2 Rotor or bucket covers are designed to minimize the possibility of fluid leakage during centrifugation Bucket covers for swinging bucket rotors help to contain fluids within the bucket in the event of tube breakage or blood bag failure Some fixed angle rotors have available dual locking lid mecha nisms that provide added biosafety by allowing the rotor to be loaded into and removed from the centrifuge with the lid in place The rotor may be placed under a safety hood
95. es centrifuges have Delrin rings on the spindle hubs to prevent sticking and do not require lubrication 3 Place the rotor in the rotor vise and use the plug wrench to remove the rotor plugs 4 Remove spacers with the floating spacer removal tool 338765 and tubes with the tube removal tool 361668 e e EU 5 Refer to Section 3 USING TUBES BOTTLES AND ACCESSO 838765 RIES for sample recovery methods 6 6 J Series Rotors and Tubes Using Vertical Tube and Rack Type Rotors USING A RACK TYPE ROTOR TRAYS AND TUBES Two kinds of trays are available for use in the rack type rotor to accommodate a variety of racks and tube sizes The trays can be identified by color as listed in the rotor manual Some racks require the use of adapters spacers or frames to ensure a proper fit in the tray Refer to the rotor manual to select compatible labware IB NOTE Tubes should be no longer than 105 mm for proper clearance ROTOR PREPARATION AND LOADING For runs at other than room temperature refrigerate or warm the rotor beforehand for fast equilibration Prerun Safety Checks Read all safety information in the rotor manual before using the rotor 1 Make sure that the rotor yoke and carriers are clean and show no signs of corrosion or cracking 2 Check the chemical compatibilities of all materials used Refer to Appendix A CHEMICAL RESISTANCES 3 Verify that the tube racks trays adapters and spacers being
96. evel for uncapped tubes as listed in the applicable rotor manual Cap requirements depend on the tube or bottle material diameter and wall thickness as well as on the rotor The applicable rotor manual specifies which cap should be used with a particular tube or bottle use of the wrong cap could cause a rotor mishap When closed containers are required several choices are available e Cap assemblies threaded caps with inserts and O rings that provide a leakproof closure to accommodate a capacity container load that is to the bottom of the insert Single piece cap assem blies have the insert permanently attached Threaded caps without inserts or O rings these are not as liquid tight as the cap assemblies therefore the meniscus must be kept lower to prevent leakage Speed reductions may also be required with lower fill volumes e Snap on caps these caps are simple to use but are not as liquid tight as the cap assemblies or threaded caps They require an even lower meniscus to prevent leakage FILLING AND CAPPING BOTTLES J Series Rotors and Tubes To prevent spillage and provide support polycarbonate and polypro pylene bottles used in fixed angle rotors must be capped when fill levels exceed the maximum level allowed for uncapped bottles Bottles should be filled to maximum fill levels when spun at full rated speeds Unless specified otherwise the minimum recommended volume for bottles is half full this will requir
97. ferential between steps and the cross sectional area allow 3 to 6 hours for diffusion at room temperature and about 16 hours at 0 to 4 C For diffusion of step gradient in Quick Seal and capped straightwall tubes slowly lay the tube on its side tube contents will not spill but make sure the tube does not roll After 2 hours at room temperature slowly set the tube upright Once the gradient is prepared layer the sample on top of the gradient For thinwall tubes only partially filled with gradient add a buffer solution to fill the tube to provide tube wall support Although the gradient volume is reduced sample volume is not changed I gt NOTE If a partially filled thickwall tube is centrifuged the tube does not require liquid support and therefore the buffer solution is not required GENERAL FILLING AND SEALING OR CAPPING REQUIREMENTS 3 2 See Table 3 1 for general filling and sealing or capping requirements for tubes and bottles used in J series rotors Maximum fill volume includes sample and gradient Refer to individual rotor manuals for specific filling and capping requirements J Series Rotors and Tubes Using Tubes Bottles and Accessories Table 3 1 General Filling and Sealing Requirements for Tubes and Bottles Polyallomer thinwall tubes thickwall tubes Quick Seal tubes within 2 to 3 mm of top at least 2 full full and heat sealed full with cap 1 2 full to max capless level or full w
98. friction Windshield The windshields are made of anodized aluminum Cannisters used in some J Lite rotors are made of lightweight Se carbon fiber epoxy composite The lightweight cannisters make the Aluminum overall rotor weight significantly lighter than a comparably sized Composite A Interface all aluminum rotor Each cannister has a sleeve washer made of Teflon and Ultem which acts as a sleeve between the cannister and the aluminum rotor body A lubricated ethylene propylene rubber O ring inside the cannister closure helps create a secondary seal during centrifugation Sleeve Washer Outside Wall Teflon is a registered trademark of E I Du Pont de Nemours amp Company Ultem is a registered trademark of GE Plastics J Series Rotors and Tubes 1 3 Rotors 1 4 _ Vent Plug DRIVE PINS gECKMAyy Drive Pins Angled pins shown pins can also be vertical or horizontal Avanti J Centrifuge Drive Spindle Assembly Transparent plastic covers are available for some swinging bucket rotor buckets to help contain spills and glass particles in the event of tube breakage The covers are made of high impact Ultem Each cover requires an O ring The O rings or gaskets in rotor assemblies with lids are made of Buna N elastomer and maintain atmospheric pressure in the rotor if they are kept clean and lightly coated with silicone vacuum grease Plug gaskets in vertical tube rotors are ma
99. fuges In addition to these instructions observe procedures and precautions provided in the applicable rotor and centrifuge manuals Refer to Section 2 for tube selection information and Section 3 for recommended filling and sealing requirements for each tube type and for sample recovery procedures Refer to Section 7 for information on the care of rotors and accessories Refer to Table 6 1 for general operating specifications for vertical tube and rack type rotors Vertical tube rotors see Figure 6 1 hold tubes parallel to the axis of rotation therefore bands separate across the diameter of the tube rather than down the length of the tube see Figure 1 3 Vertical tube rotors are useful for separating and banding subcellular particles These rotors have plugs that are screwed into the rotor cavities over sealed Quick Seal tubes The plugs with spacers when required restrain the tubes in the cavities and provide support against the hydrostatic force generated by centrifugation Refer to Section 3 for information about filling and sealing Quick Seal tubes for use in vertical tube rotors Using Vertical Tube and Rack Type Rotors Table 6 1 General Specifications for Beckman Coulter J Series Vertical Tube and Rack Type Rotors JV 20 20 000 rpm 600 93 80 67 8 x 39 mL 312 mL 0 Angle 41619xg to 206 800 JR 3 2 3200 rpm 600 199 80 67 320 x 1 mL 320 mL 90 Angle 2280 x g to at Speed 25 606 800
100. fully especially if this is your first experience with Beckman Coulter products Section 1 describes by usage Beckman Coulter s currently produced J series rotors this should help you determine the appropriate rotor to use for a particular application Also included in this section is a discussion of rotor materials components and centrifugation techniques Section 2 describes various tubes adapters spacers and cannisters to help you choose a particular container for your application Section 3 provides instructions for using tubes bottles cannisters and related accessories Section 4 contains step by step procedures for preparing a fixed angle rotor for a centrifuge run Similar information is available for swinging bucket rotors in Section 5 and Section 6 contains the same type of information for vertical tube and rack type rotors Elutriation zonal and continuous flow rotors are not covered in this manual e Section 7 provides rotor tube and accessory care and maintenance information as well as some diagnostic hints Please read it Proper rotor care results in longer rotor life Several appendixes contain information that may be of special interest Appendix A lists chemical resistances for rotor and accessory materials to help determine compatibility with a variety of solutions Appendix B contains Temperature Compensation Tables for various rotors Appendix C contains reference information on some
101. he JR 3 2 rotor gt NOTE These rotors are not used in Avanti J series centrifuges J Series Rotors and Tubes Using Vertical Tube and Rack Type Rotors INSTALLING THE ROTOR N CAUTION The centrifuge drive spindle can be bent or broken if the rotor is forced sideways or dropped onto it Install the rotor by centering it over the spindle and carefully lowering it straight down 1 Liftthe rotor by the yoke and carefully lower it straight down onto the drive spindle Rotor F Drive Pins 2 2 Rotate the rotor by hand until the drive pins seat on the drive a spindle hub Grooves 4 SN Hub NS Drive Spindle AsserbIy In older model centrifuges be sure the pins in the rotor drive Older Model Centrifuges hole are located in the grooves of the drive spindle hub Teeth LUE NS Drive In newer model centrifuges be sure the pins in the rotor drive NN hole are not sitting on top of the teeth on the drive spindle hub Newer Model Centrifuges A CAUTION The pins located in the rotor hub must be seated correctly on the centrifuge drive spindle Running a rotor that is not seated properly may result in rotor failure 3 Turn the tie down knob to the right clockwise until the rotor is secure J Series Rotors and Tubes 6 9 Using Vertical Tube and Rack Type Rotors REMOVAL AND SAMPLE RECOVERY AN CAUTION If disassembly reveals evidence of leakage you should assume that some fluid escaped the container or rotor Ap
102. he rotor with the tie down screw and tighten the screw with the torque bar or tie down tool If the rotor is left in the centrifuge between runs ensure that the screw is tight before each run 5 7 Using Swinging Bucket Rotors 5 8 Tie down Tool 368521 e JS 4 2A and JS 4 28MA secure the rotor to the drive hub with Tie down Bolt the tie down bolt 368518 Tighten the bolt with the tie down tool ae 368521 then remove the tool IB NOTE Older JS 4 2A or JS 4 2SMA rotors may be secured to the drive hub with tie down bolt 339031 Tighten the bolt with the spanner poo wrench 870666 or tie down tool 368521 Bolt 339031 then remove the wrench or tool Spanner Wrench 870666 f e JS 5 0 secure the rotor to the drive hub with the tie down bolt 367824 Tighten the bolt with the tie down tool 368521 then remove the tool Loading JS 24 Series Rotors 1 To install the rotor carefully lift it up off the rotor stand with both hands do not lift the rotor by the adapter and place it on the drive hub Make sure that the rotor pins are perpendicular to the drive hub pins The pins must not rest on top of each other turn the rotor to the right clockwise by hand to check for proper installation 2 Turn the tie down knob to the right clockwise to secure the rotor SYMMETRIC AND BALANCED LOADING To ensure optimal performance and stability swinging bucket rotors must be loaded symmetrically
103. hen be heat sealed for pellet storage or disposal 2 13 3 Using Tubes Bottles and Accessories This section contains general instructions for filling and capping the labware used in Beckman Coulter J series rotors for selecting and using the appropriate accessories and for recovering samples after a run Individual rotor manuals provide specific instructions on tubes bottles and accessories that can be used in a particular rotor 1 Rotor use instructions are in Section 4 for fixed angle rotors in Section 5 for swinging bucket rotors and in Section 6 for vertical tube and rack type rotors A table of chemical resistances is in Appendix A of this manual Reference information on some commonly used gradient materials is in Appendix C GRADIENT PREPARATION Many commercial gradient formers are available These devices usually load a tube by allowing the gradient solutions to run down the side of the tube The heaviest concentration is loaded first followed by successively lighter concentrations This method is acceptable for wettable tubes however loading a nonwettable tube such as Ultra 5 Clear polyallomer and polycarbonate by allowing solutions to run down the side of the tube can cause mixing Added First 10 Gradients in nonwettable tubes can be prepared using a gradient 15 former by placing a long syringe needle or tubing to the tube bottom and reversing the gradient chambers In that way the lightest gradie
104. ifferential rates of sedimentation while others may reach their isopycnic point in the gradient In most cases when banding two or three components by rate zonal separation run times can be shortened considerably if reduced fill levels are used Tubes are partially filled with gradient but the sample volume is not changed however gradient capacity will be reduced Thickwall tubes should be used for this technique since thinwall tubes will collapse if not full J Series Rotors and Tubes Rotors BLOOD COMPONENT SEPARATIONS Centrifugation is the primary method for processing blood because it provides the required high throughput reproducibility and versatility Most blood components can be separated in one or two runs Gener ally two types of runs are performed Soft spin runs short centrifugation runs 3 to 5 minutes at low g forces 2000 to 3000 x g at ambient temperature are used to keep small cells or platelets in suspension while the larger cells sediment This type of run is used to obtain platelet rich plasma and red blood cell concentrate from whole blood Hard spin runs are longer 5 to 7 minutes at higher g forces 4000 to 5000 x g at ambient temperatures or at 4 C and are used to separate fresh plasma from cellular components Soft spin and hard spin techniques are often combined Refer to Appendix D for further information about separation of blood components by centrifugation GENERAL OPERATING INF
105. ightly but evenly lubricated with Spinkote lubricant 306812 2 Ensure that O rings are in good condition and are lightly but evenly coated with silicone vacuum grease 335148 3 Ensure that all sealing surfaces are smooth and undamaged for proper sealing 4 Before each use of the rotor make sure that rotor pins and bucket pin sockets are lubricated with Tri flow oil 883371 use Paint On Graphite Lubricant 977212 on pin sockets for JS 5 9 JS 5 3 JS 5 0 and JS 4 3 rotors Special Preparation Instructions for JS 24 Series Rotors pus Hanger Bucket Place the rotor on the rotor stand 362785 when it is not in the centrifuge 1 Load the filled containers into the buckets Complete loading by placing the correct floating spacers if required over the tubes 2 Ensure that bucket O rings are lightly but evenly coated with silicone vacuum grease Do not run a bucket without an O ring as the bucket will leak 3 Be sure that metal threads in the bucket caps are clean and lightly but evenly lubricated with Spinkote lubricant Put bucket caps on the buckets and screw them down manually 4 Hook all buckets loaded or empty on to the rotor and be sure that both hooks are on the crossbar All six buckets must be in the same size Do not intermix the smaller and larger buckets in a single run If fewer than six tubes are being run they must be arranged symmetrically in the rotor Opposing tubes must be filled t
106. il Tables Table 1 1 Rotors Used in Beckman Coulter J Series Centrifuges 1 5 Table 2 1 Characteristics and Chemical Resistances of Tube and Bottle Materials cc RR hh 2 2 Table 3 1 General Filling and Sealing Requirements for Tubes and Bottles sss mios een eec es ledere e Ro a og 3 3 Table 3 2 Aerosolve Tube Racks 0 0 cece eee een eee 3 18 Table 4 1 General Specifications for Beckman Coulter J Series Fixed Angle Rotors 605 wees OLD O9 eb Rad Rs 4 3 Table 5 1 General Specifications for Beckman Coulter J Series Swinging Bucket Rotors 1 0 0 0 cece eee eee 5 3 Table 5 2 Microplate Carriers Used with J6 Series Rotors 5 16 Table 6 1 General Specifications for Beckman Coulter J Series Vertical Tube and Rack Type Rotors 0 0 cece ee eee eee eee 6 2 Table 7 1 Tube and Bottle Sterilization and Disinfection 7 8 Table B 1 Temperature Compensation Settings for the J2 HC Centrifuge B 2 Table B 2 Temperature Compensation Settings for the J2 21 J2 21B J2 21C and J2 HS Centrifuges 0 0 0 0 00 00000 B 4 Table B 3 Temperature Compensation Settings for the J2 MI J2 21M J2 MC and J2 21M E Centrifuges 0 0 0 eee eee eee B 6 Table B 4 Temperature Compensation Settings for the J6 Centrifuges B 8 Table C 1 Commonly Used Gradient Materials with Their Solvents C 2 Table C 2 Density Refractive Index and Co
107. il lids 538619 have a bioinert adhesive backing enabling complete plate sealing The lids are presized for multiwell deep well and square well plates and cause no reduction in well capacity A 4 inch soft rubber roller 538618 is required to ensure secure sealing of the foil lids Tubes and bottles used with adapters can be filled and capped if applicable according to the type of container and the design of the rotor being used J Series Rotors and Tubes Using Tubes Bottles and Accessories USING SOLID MULTITUBE ADAPTERS The solid adapters available in several tube configurations can be filled and loaded into rotor buckets or cavities without any prepara tion They can also be used as tube racks in the laboratory USING MODULAR DISK MULTITUBE ADAPTERS These adapters can also be used as tube racks in the laboratory The adapter disks are color coded by the tube size they accom modate the number of disks used in an adapter assembly depends upon the length of tubes used Refer to the applicable rotor manual to determine the kind of adapter required for the tubes you are using Assemble modular disk adapters as follows 1 Select the appropriate adapter base and attach a bail to it 2 Place the base and bail in an empty bucket or on the lab bench not in the rotor 3 Position one of the disks so that its grooves are aligned with the bail Push the disk down until the bail snaps into the grooves 4 Add more
108. ing acceleration the rotor shifts so as to rotate about its center of mass While passing through this speed range the rotor will usually vibrate Do not set operating speeds that are within a rotor s critical speed range as listed in the rotor manual Minimum Speeds Some buckets or carriers will not achieve their full horizontal position if the rotor is run below minimum rotating speed Refer to the individual rotor manual for speed requirements 1 18 J Series Rotors and Tubes Rotors TEMPERATURE COMPENSATION TEMP C Lt Avanti J Series Centrifuges Analog J2 Series Centrifuges SET O COMP J Series Rotors and Tubes To ensure that the rotor reaches the required temperature during centrifugation some temperature compensation may be required because of the mass of these rotors Tables listing temperature compensation units for various rotors are contained in Appendix B and individual rotor manuals Follow the instructions below for the model of centrifuge being used Avanti J series centrifuges provide automatic temperature compensa tion Enter the run temperature according to the instructions in your centrifuge instruction manual No additional input is required Set temperature compensation in analog J2 model centrifuges models J2 HS J2 21 and J2 HC as follows 1 Turn the SET knob on the centrifuge panel to the required sample temperature 2 Find the compensation value in Appendix B or in th
109. ionation or separation into components in order to use collected blood most efficiently Termed component therapy multiple use of different parts of the blood helps conserve this scarce resource and allows patients to receive only the components they need As shelf life and storage requirements vary conditions can be optimized by separating components In addition to collection of whole blood for separation into components several techniques involve collection of whole blood separation collection of a fraction for infusion and return of the remainder to the donor n plasmapheresis a unit of blood is taken to obtain plasma sepa rated and the red cells are immediately reinfused to the donor Most plasmapheresis is performed for source plasma which is not intended for intravenous transfusion but separated by large scale fractionation into clotting factors especially factor VIII albumin and specific immunoglobulins During plateletpheresis whole blood is collected platelets sepa rated via centrifugations and platelet depleted red blood cells returned to the donor The plasma may be returned to the donor or collected for fractionation into clotting factors and albumin Blood Component Separation Leukapheresis is the separation of leukocytes or white blood cells from whole blood The leukocyte depleted and platelet depleted red blood cells are continuously or intermittently returned to the donor COMPONE
110. ith cap full and heat sealed not used not used full and heat sealed bottles min to max see rotor 1 2 full to max see rotor manual with not used manual with screw on Screw on cap or cap assembly cap or cap assembly Ultra Clear open top tubes within 2 to 3 mm of top full with cap not used Quick Seal tubes not used full and heat sealed full and heat sealed Polycarbonate thickwall tubes at least 1 2 full 1 2 full to max capless level or full not used with cap bottles at least 1 2 full min to max see rotor manual with not used screw on Cap or cap assembly Stainless Steel tubes any level any level with cap or cap assembly not used Polypropylene tubes and bottles at least 1 2 full 1 2 full to max capless level or full not used with cap or cap assembly Polyethylene tubes at least 1 2 full 1 2 full to max capless level or full not used with cap Cellulose Propionate tubes and bottles at least 1 2 full 1 2 full to max capless level not used J Series Rotors and Tubes 3 3 Using Tubes Bottles and Accessories WORKING WITH PHYSIOLOGICAL FLUIDS N WARNING Handle body fluids with care because they ony can transmit disease No known test offers ey complete assurance that they are free of micro organisms Some of the most virulent Hepatitis B and C and HIV I V viruses atypical mycobacteria and certain systemic fungi further emphasize the need for aerosol protection Handle oth
111. ixed Angle Rotors continued JA 20 20 000 rpm 600 to 108 70 32 8x50 mL 400 mL 34 Angle 48 400 x g 800 770 JA 18 1 18 000 rpm4 600 to 24 x 1 8 mL 43 2 mL i 45 Angle 42 100x g 800 116 105 95 Adapter 156 or 25 Angle 17 000 rom4 112 106 101 24 x 1 8 mL 43 2 mL Adapter 36 300 x g 91 JA 18 18 000 rome 600 to 132 98 64 10 x 100 mL 1 liter 23 Angle 47 900 x g 800 566 JA 17 17 000 rpmf 600 to 123 90 56 14x50 mL 700 mL 25 Angle 39 800 x g 800 690 JLA 16 250 16 000 rpms 600 to 134 90 46 6 x 250 mL 1 5 liter 25 Angle 38 400 x g 800 1 090 Continued a Maximum speeds are based on a solution density of 1 2 g mL in all rotors except for the JA 18 1 which is rated for a density of 1 4 g mL Critical speed range is the range of speeds over which the rotor shifts so as to rotate about its center of mass Passing through or running at the critical speed range is characterized by some vibration Maximum speed in an Avanti J E centrifuge is 21 000 rpm 4 When a JA 18 1 rotor is used in the J2 HC centrifuge derate the rotor as follows when the 45 adapters are used do not run the rotor above 15 000 rpm when 25 adapters are used do not run the rotor above 16 000 rpm Maximum speed in an Avanti J series centrifuge except Avanti J E maximum speed in an Avanti J E for a rotor with magnets maximum speed for rotor
112. izontal position as the rotor accelerates 15 J Series Rotors and Tubes Rotors Tubes in vertical tube rotors designated JV are held parallel to the axis of rotation These rotors have plugs screwed into the rotor cavities over sealed tubes that keep the tubes in the cavities and provide support for the hydrostatic forces generated by centrifugation Tubes in the rack type rotor designated JR are held in gamma counter racks Racks are loaded into special plastic trays which are then loaded into carriers at a resting angle During centrifugation the carriers swing out to a completely horizontal position MATERIALS Most Beckman Coulter J series rotors are made of aluminum and are anodized to protect the metal from corrosion The JS 13 1 and JS 7 5 rotors are painted with polyurethane paint and are not anodized The anodized coating is a thin hard layer of aluminum oxide formed electrochemically in the final stages of rotor fabrication A black or colored dye may be applied over the oxide for rotor family identifica tion The coating can be damaged if careful cleaning procedures are not followed Therefore it is especially important to clean aluminum rotors with brushes that will not scratch the anodized coating and to use a noncorrosive neutral pH detergent Refer to Section 7 for 2 i cleaning and maintenance procedures A Some J series rotors have attached windshields to reduce air
113. lder it may cause the tube to leak I gt NOTE It is very important to apply the heat sink imme diately To do so we recommend that you have it in one hand ready to apply as soon as needed c Remove the Tube Topper IMMEDIATELY place the large end of the heat sink over the seal former Hold it there for a few seconds while the plastic cools do NOT let the seal former pop up If the seal former does pop up the tube may not have an adequate seal and may need to be resealed d Remove the heat sink When the seal former cools remove it by hand or with the removal tool 361668 4 After completing either heat sealing method squeeze the tube gently if the tube contents may be disturbed to test the seal for leaks If the tube does leak try resealing it using Method A 5 The tube is now ready for centrifugation Seal the remaining tubes 6 Return the Tube Topper to its charging stand when finished Using Tubes Bottles and Accessories CAPPING MULTIWELL TITER PLATES Cap Strips 96 Cap Strip Aluminum Foil Lids Roller 538618 Seal amp Sample Foil Lid USING ADAPTERS Multiwell titer plates regular deep well and square well can be run uncovered or using one of the available cover types Available sterile or nonsterile 96 cap strips can be used with deep well plates When these caps are used the capacity of each well is reduced to 1 0 mL Seal amp Sample aluminum fo
114. les and Accessories 2 10 MULTITUBE ADAPTERS Solid Multitube Adapters Adapters are used to enable centrifugation of multiple tubes in the bucket of a swinging bucket rotor or in a fixed angle tube cavity These solid adapters available in several tube configurations are made of Ultem Noryl polypropylene or aluminum that is anodized for corrosion protection They can be filled and loaded into rotor buckets or cavities without any preparation These adapters can also be used as tube racks in the laboratory These adapters can also be used as tube racks in the laboratory The adapter disks are color coded by the tube size they accommodate the number of disks used in an adapter assembly depends upon the length of tubes used Refer to the applicable rotor manual to determine the kind of adapter required for the tubes you are using A tube decanter is available to hold tubes securely in some adapters allowing all tubes to be decanted at once J Series Rotors and Tubes Tubes Bottles and Accessories Do not intermix modular adapters or their individual parts from Beckman Coulter s J6 series rotors with those for the JS 4 3 rotor While the adapters are similar in appearance they have very different weights J6 adapters have bails vertical supports that are curved at the top bails for the JS 4 3 adapters are straight Keep J6 and JS 4 3 disks and bases separate from each other mixing them can caus
115. lied with it J Series Rotors and Tubes Seal Former Using Tubes Bottles and Accessories Remove the Tube Topper from the charging stand Leave the pushbutton turned to LOCK position N WARNING Touching the heated tip of the Tube Topper will cause burns When the pushbutton is pressed the tip heats almost immediately Make sure the pushbutton is turned to LOCK position unless you are actually sealing a tube 2 Place a seal former on each tube stem The Teflon coating on the seal formers is permanent Do not scratch the interior of the formers as you may damage this coating 3 Sealeach tube using Method A or B Method A is preferable when sealing smaller tubes or when resealing a tube that leaks N CAUTION Always keep the Tube Topper in its charging stand when not in use Do not lay the unit against any surface after use until the tip has cooled 3 to 5 minutes after shut off Teflon is a registered trademark of E I Du Pont de Nemours amp Co J Series Rotors and Tubes Using Tubes Bottles and Accessories METHOD A WITH THE SEAL GUIDE Seal Guide a Place a seal guide with the flat side down over the seal G3 former y b Turn the Tube Topper pushbutton to USE position Press the pushbutton and wait 3 to 5 seconds for the tip to heat c Apply the tip of the Tube Topper vertically to the seal former Press down gently for about 10 seconds The seal guide should move down the tube stem u
116. ll volumes listed in the applicable rotor manual Ultra Clear For swinging bucket rotors fill to within 2 or 3 mm of the top of the tube Refer to the applicable rotor manual Polypropylene Polyethylene Stainless Steel Fill all opposing tubes to the same level For swinging bucket rotors fill to within 2 or 3 mm of the top of the tube Hill thickwall polypropylene tubes at least half full to maximum level in fixed angle rotors Speed reduction is required Refer to the applicable rotor manual For swinging bucket and fixed angle rotors fill these tubes from half full to maximum level Refer to the applicable rotor manual Because of their strength stainless steel tubes can be centrifuged while filled to any level with all opposing tubes filled to the same level However run speeds must be reduced due to their weight The criteria for speed reduction depends on the tube cap material and the strength of the rotor being used Refer to the applicable rotor manual or Run Speeds for Stainless Steel Tubes publication L5 TB 072 for correct run speeds J Series Rotors and Tubes CAPPING TUBES 60g D0 J DO Using Tubes Bottles and Accessories Caps must be used with thinwall polyallomer and Ultra Clear tubes in fixed angle rotors To prevent spillage thickwall polyallomer poly carbonate and stainless steel tubes must be capped when fill levels exceed the maximum l
117. lly seated on 3 8 J Series Rotors and Tubes Using Tubes Bottles and Accessories the cup Use green gaskets 369261 if you are using liners Use red gaskets 369257 if you are using cups alone with or without partitions 3 Place a liner in each cup if applicable 4 You may fill the cups now or close the lid and fill through the spout gt NOTE If liners are not used partitions 369259 may be inserted into the slots inside the cups Remove the red cup gaskets 369257 before inserting partitions and be sure to reinstall the gaskets 5 Place a cover on each cup and fasten the latch securely If the latch will not fasten check to make sure that the gasket is properly installed The latch cannot be fastened if the gasket is not fully seated Be sure that the latch is fastened before lifting the cup by the handle 6 Ifthe cups were not filled previously load sample into each cup through the cover spout using a funnel tubing 1 27 cm V 2 in O D or a pipette Use the fill line indicators to assist in filling all four cups to the same level All four cups must balance to within 25 grams of each other When loading is complete snap a plug into place in each cover spout FILLING AND SEALING QUICK SEAL TUBES Fill each tube to the base of the neck using a syringe with a 13 gauge or smaller needle A small air space no larger than 3 mm may be left but an air bubble that is too large can cause the t
118. lubrication Do not use sharp tools on a rotor as the surface can get scratched Corrosion begins in scratches and may open fissures in the rotor with continued use The corrosion process accelerates with speed induced stresses The potential for damage from corrosion is greatest in aluminum rotors and components 7 1 Care and Maintenance 7 2 CLEANING Rotor Cleaning Kit 339558 Wash rotors and rotor components immediately if salts or other corrosive materials are used or if spillage has occurred DO NOT allow corrosive materials to dry on the rotor IB NOTE Do not wash rotor components or accessories in a dishwasher Do not soak in detergent solution for long periods such as overnight With normal usage wash rotors frequently to prevent corrosion that can begin in scratches Remove buckets from yokes before cleaning swinging bucket rotors N CAUTION Do not immerse or spray a swinging bucket rotor yoke or body with water because liquid can become trapped in the hinge pin area and lead to corrosion 1 Use plastic or wooden tools to remove O rings or gaskets for cleaning do not use metal tools that could scratch anodized surfaces Use a mild detergent such as Beckman Solution 555 339555 diluted 10 to 1 with water and a soft brush to wash rotors and rotor components and accessories Most laboratory detergents are too harsh for aluminum rotors and components The Rotor Cleaning Kit 339558 contains
119. lypropylene The tubes are placed in adapters for use in some fixed angle rotors They are also used in multitube adapters in the buckets or carriers of swinging bucket rotors The number and arrangement of tubes in opposing adapters should be balanced J Series Rotors and Tubes Tubes Bottles and Accessories BOTTLES Bottles are available in polycarbonate hard and clear polypropylene translucent and polyallomer translucent Sealed polyallomer or polycarbonate bottles available for most fixed angle rotors have a three piece liquid tight cap assembly Other bottles have screw on caps Cap assemblies should always be removed before autoclaving bottles Bottle selection depends on the rotor in use and the specific application refer to the applicable rotor manual JLA 8 1000 and JLA 9 1000 J Lite rotors run only the specially nad O ring designed Beckman Coulter bottles and accessories with poly AutoVent Plug O ring carbonate plug seals A Radel cap closure placed over the plug l __ AutoVent Plug screws onto the bottle During centrifugation the cap closure is Sese HE wen pulled down into the cannister creating a tight seal The cap closure provides secondary containment of the sample in the event of bottle ied oe ee leakage O rings on the plug and the cap closure create the seals ll polycarbonate the O rings must be free of defects dry and unlubricated to ensure or polypropylene sealing
120. me f in hours required for pelleting ped 3 where s is the sedimentation coefficient of the particle of interest in Svedberg units Because s values in seconds are such small numbers they are generally expressed in Svedberg units S where 1 S is equal to 10 3 seconds It is usual practice to use the standard sedimenta tion coefficient s20 based on sedimentation in water at 20 C Clearing factors can be calculated at speeds other than maximum rated speed by use of the following formula 2 ptor speed of d 4 adj actual run speed 4 s dr dt x Vor where dr dt is the sedimentation velocity J Series Rotors and Tubes Rotors Run times can also be calculated from data established in prior experiments when the k factor of the previous rotor is known For any two rotors a and b iol 5 where the k factors have been adjusted for the actual run speed used Figure 1 3 lists sedimentation coefficients for some common biological materials The k factors at rated speeds for Beckman Coulter J series rotors are provided in the table of general specifica tions in each rotor use section Cytochrome c Collagen Yeast tRNA Albumin Soluble Proteins Luteinizing hormone Immunoglobulin G Aldolase Nucleic Acids Catalase E coli rRNA a Macroglobulin Z Calf liver DNA Ribosomal subunits 40 Vesicular stomatitis
121. me it is placed in the freezer SEPARATION OF BLOOD COMPONENTS BY CENTRIFUGATION Centrifugation is the primary method for processing blood because it offers the required high throughput reproducibility and versatility Most blood components can be separated in one or two runs Generally two types of centrifugation runs are performed see Figure D 1 Soft spin runs short centrifugation runs 3 to 5 minutes at low g forces 2000 to 3000 x g at ambient temperature are used to keep small cells or platelets in suspension while the larger cells sedi ment This type of run is used to obtain platelet rich plasma and red blood cell concentrate from whole blood Hard spin runs are longer 5 to 7 minutes at higher g forces 4000 to 5000 x g at ambient temperature or at 4 C and are used to separate fresh plasma from cellular components Soft spin and hard spin tech niques are often combined Donor blood is collected in plastic bags with one or more satellite bags double triple or quad packs containing anticoagulant and preservative After each centrifugation run the sedimented fraction is squeezed into its respective satellite bag Common anticoagulants and preservatives include citrate phosphate dextrose CPD citrate phosphate dextrose citrate citric acid adenine CPDA 1 saline adenine glucose minitol SAG M and dextrose sodium chloride mannitol adenine ADSOL Blood separations occur during centrifugation because of
122. n fixed angle rotors Tubes Bottles and Accessories ULTRA CLEAR TUBES STAINLESS STEEL TUBES MICROFUGE TUBES Ultra Clear tubes made of a tough thermoplastic are thinwall and not wettable but can be made wettable see Section 3 Ultra Clear tubes are available in two types open top and Quick Seal They are transparent centrifuge tubes offering easy location of visible banded samples Standard straight wall Ultra Clear tubes must be filled completely and capped for use in fixed angle rotors Ultra Clear tubes which can be used one time only have good resis tance to most weak acids and some weak bases but are unsatisfactory for DMSO and most organic solvents including all alcohols Ultra Clear tubes should not be autoclaved Stainless steel tubes offer excellent resistance to organic solvents and heat but should not be used with most acids or bases They offer only marginal resistance to most gradient forming materials other than sucrose and glycerol Stainless steel tubes are very strong and can be centrifuged when filled to any level Because of their weight however run speeds must often be reduced see publication L5 TB 072 Stainless steel tubes can be used indefinitely if they are undamaged and not allowed to corrode They may be autoclaved after use as long as they are thoroughly dried before storage Microfuge tubes 1 5 mL tubes with attached caps are made of clear polyallomer or of clear or colored po
123. nal S A 22 Rue Juste Olivier Case Postale 301 303 CH 1260 Nyon Switzerland Telephone 41 22 994 07 07 Fax 41 22 994 07 00 FRANCE Beckman Coulter France S A Paris Nord II 33 66 rue des Vanesses B P 50359 Villepinte France 95942 ROISSY CDG Cedex Telephone 01 49 90 90 00 Fax 01 49 90 90 10 e mail bioresearch france 2 beckman com GERMANY Beckman Coulter GmbH Siemensstrasse 1 D 85716 Unterschleissheim Lohhof Germany Telephone 89 35870 0 Fax 89 35870 490 e mail bioresearch de beckman com Qoo COULTER Beckman Coulter Inc HONG KONG Beckman Coulter Hong Kong Ltd 12th Floor Oxford House 979 King s Road Taikoo Place Hong Kong Telephone 852 2814 7431 2814 0481 Fax 852 2814 1599 2873 4511 ITALY Beckman Coulter S p a Centro Direzionale Lombardo Palazzo F 1 Via Roma 108 20060 Cassina de Pecchi Milano Italy Telephone 02 953921 Fax 02 95392264 JAPAN Beckman Coulter K K Toranomon 37 Mori Bldg 3 5 1 Toranomon Minato ku Tokyo 105 0001 Japan Telephone 03 5404 8359 Fax 03 5404 8436 MEXICO Beckman Coulter de Mexico S A DE C V Avenida Popocat petl 396 Colonia Gral Pedro Maria Anaya Codigo Postal 03340 Maxico D F Mexico City Telephone 525 605 77 70 Fax 525 575 18 25 NETHERLANDS Beckman Coulter Nederland B V Nijverheidsweg 21 3641 RP Mijdrecht Postbus 47 3640 AA Mijdrecht The Netherlands Telephone 0297 230630 Fax
124. ncentration Data Cesium Chloride at 25 C Molecular Weight 168 37 C 3 Table C 3 Density Refractive Index and Concentration Data Sucrose at 20 C Molecular Weight 342 3 00 004 C 4 Table C 4 Density Conversion for Cesium and Rubidium Salts at 20 C C 5 Table D 1 Blood Component Storage 0 0 eee D 3 Table D 2 Blood Bank Methods 0 0 0 0 cece eee eee D 5 Vili J Series Rotors and Tubes 1 Rotors This section is an introduction to the Beckman Coulter family of J series rotors providing general information on rotor design selection and operation Rotor designs described are fixed angle swinging bucket vertical tube and rack type Specific instructions for using each type of rotor are contained in Sections 4 through 6 Care and maintenance information for all of these rotors is contained in Section 7 Elutriator continuous flow and zonal rotors are not covered in this manual The elutriator rotors are described in detail in their respective rotor instruction manuals publications JE6B IM and JE5 IM the continuous flow zonal rotor JCF Z is described in publication JCFZ IM GENERAL DESCRIPTION ROTOR DESIGNATIONS J Series Rotors and Tubes Beckman Coulter J series rotors are usually named according to the type of rotor and the rotor s maximum allowable revolutions per minute in thousands referred to as rated speed For example the JA 12 roto
125. nd Section 3 for recommended labware filling and sealing requirements and for sample recovery procedures Refer to Section 7 for information on the care of rotors and accessories Swinging bucket rotors see Figure 5 1 are normally used for density gradient separations either isopycnic or rate zonal Refer to Table 5 1 for general rotor specifications A tie down device or lid locking knob is used to lock the rotor to the centrifuge drive hub before the run begins Tubes or bottles in swinging bucket rotors are held in the rotor buckets that are attached to the rotor body by hinge pins The buckets swing out to horizontal position as the rotor is accelerated and stay horizontal until rotor deceleration begins During deceleration the buckets gradually return to vertical position III NOTE Although rotor components and accessories made by other manufacturers may fit in the Beckman Coulter rotor you are using their safety in the rotor cannot be ascertained by Beckman Coulter Use of other manufacturers components or accessories in the rotor may void the rotor warranty and should be prohibited by your laboratory safety officer Only the components and accessories listed in the applicable rotor manual should be used 5 1 Using Swinging Bucket Rotors JS 7 5 Axis of Rotation Axis of Rotation JS 4 3 Axis of Rotation
126. ng the cardiovascular system with extra fluid Blood Component Separation volume are commonly administered to treat anemia LEUKOCYTE DEPLETED RED BLOOD CELLS These are prepared by removing most leukocytes and platelets from fresh whole blood Obtained by differential separation they are given to recipients with antileukocyte antibodies to prevent adverse reactions SHELF LIFE Table D 1 lists approximate storage times for the separated components Table D 1 Blood Component Storage Approximate Component Storage Life RBC with ADSOL 42 days RBC with SAG M 35 days RBC with CPDA 1 25 days RBC without preservative 21 days RBC frozen without addition of 10 days cryoprotective agent Plasma frozen 1 year ADSOL dextrose sodium chloride mannitol adenine SAG M saline adenine glucose minitol CPDA 1 citrate phosphate dextrose citrate citric acid adenine J Series Rotors and Tubes D 3 Blood Component Separation FREEZING A cryoprotective agent such as glycerol can be added to extend the life of frozen red blood cells These cells can then be stored for up to 3 years at 80 C Prior to use the cells are thawed and the glycerol is removed by washing If the plasma will not be separated within 15 hours of collection it must be frozen within 6 hours of collection Freezing must be carried out in a flash freezer with complete freezing accomplished within 1 or 2 hours of the ti
127. nt 20 concentration is loaded first underlayed by increasingly heavier concentrations Added Last 1 A complete list of tubes bottles and adapters is provided in the latest edition of the Beckman Coulter High Performance High Speed High Capacity Rotors Tubes amp Accessories catalog BR 8102 available at www beckmancoulter com It has been reported however that polyallomer tubes have been made wettable by soaking them in a chromic acid bath for about 30 minutes see Preparation of Polyallomer Centrifuge Tubes for Density Gradients Anal Biochem 32 334 339 H Wallace 1969 Also a method of making Ultra Clear tubes wettable that has proven successful for some users is described at the end of this section J Series Rotors and Tubes 3 1 Using Tubes Bottles and Accessories 1 mL Syringe __ 20 22 Gauge Needle Su N 2 3 mm EZ t Gradient 2 3 mm Buffer Sample with Gradient 2 to 3 Sucrose Added You can also prepare preformed step gradients by hand using a pipette Carefully layer solutions of decreasing concentration by placing the tip of the pipette at the angle formed by the tube wall and the meniscus or float the lighter gradient concentrations up by adding increased density solutions to the tube bottom using a hypodermic syringe with a long needle such as a pipetting needle Another way to form a linear gradient is to allow a step gradient to diffuse to linearity Depending on the concentration dif
128. ntil it rests on the tube shoulder Using the seal guide prevents the seal former from being pressed into the tube shoulder I gt NOTE Always apply the tip of the Tube Topper verti cally to the seal former Apply gentle pressure when sealing the tube Heat Sink d When the seal guide has moved to the correct position remove the Tube Topper and pinch the circular seal guide to hold the seal former in place e Place the heat sink small end over the cap for 2 to 3 seconds while the plastic cools do NOT let the seal former pop up If the seal former does pop up the tube may not have an adequate seal and may need to be resealed f Remove the heat sink and seal guide When the seal former cools remove it by hand or with the removal tool 361668 Save the seal guide and former for future use 3 12 J Series Rotors and Tubes Using Tubes Bottles and Accessories METHOD B WITHOUT THE SEAL GUIDE Immediately J Series Rotors and Tubes I gt NOTE Always apply the tip of the Tube Topper verti cally to the seal former Apply gentle pressure when sealing the tube a Turn the Tube Topper pushbutton to USE position Press the pushbutton and wait 3 to 5 seconds for the tip to heat b Apply the tip of the Tube Topper vertically to the seal former The seal former should move down the tube stem until it just rests on the tube shoulder Be careful NOT to press the seal former into the tube shou
129. o the same level with liquid of the same density J Series Rotors and Tubes Using Swinging Bucket Rotors LOADING THE ROTOR YOKE N CAUTION The centrifuge drive spindle can be bent or broken if the rotor is forced sideways or dropped Bos onto it Install the rotor by centering it over the Drive Pins 2 spindle and carefully lowering it straight down Hub Grooves 4 C Hub im 1 Carefully lower the rotor yoke straight down onto the drive eres spindle Rotate it by hand until the drive pins seat on the drive Assembly Older Model Centrifuges Teeth 2 em I Drive Spindle Assembly Newer Model Centrifuges _ Tie down Knob Rotor Tie down Screw Torque Bar J Series Rotors and Tubes spindle hub IB NOTE Except for the JS 24 38 and JS 24 15 rotors you can leave the rotor yoke in the centrifuge between runs unless spillage has occurred in which case you should remove the buckets or carriers and yoke and clean the centrifuge and rotor components immediately according to the instructions in the centrifuge and rotor instruc tion manuals The JS 24 38 and JS 24 15 rotors must be removed from the centrifuge to install or remove buckets When the yoke is correctly seated secure it to the drive spindle hub Rotors with tie down knobs hand tighten the tie down knob If the rotor is left in the centrifuge between runs tighten the knob before each run Rotors without tie down knobs secure t
130. omponents and accessories made by other manufacturers cannot be ascertained by Beckman Coulter Use of other manufacturers components or accessories in Beckman Coulter rotors may void the rotor warranty and should be prohibited by your laboratory safety officer High speed rotors are designed for use at the speeds indicated however speed reductions may be required because of weight considerations of tubes adapters and or the density of the solution being centrifuged Be sure to observe the instruc tions in the applicable rotor manual NEVER attempt to slow or stop a rotor by hand The strength of containers can vary between lots and will depend on handling and usage We highly recommend that you pretest them in the rotor using buffer or gradient of equivalent density to the intended sample solution to determine optimal operating conditions Scratches even microscopic ones significantly weaken glass and polycarbonate containers To help prevent premature failures or hazards by detecting stress corrosion metal fatigue wear or damage to anodized coatings and to instruct laboratory personnel in the proper care of rotors Beckman Coulter offers the Field Rotor Inspection Program FRIP This program involves a visit to your laboratory by a specially trained Beckman Coulter representative who will inspect all of your rotors for corro sion or damage The representative will recommend repair or replacement of at risk rotors to prev
131. ontaining a liquid of the same density as opposing tubes Layera low density immiscible liquid such as mineral oil on top of the sample to fill opposing tubes to the same level eg hes Figure 1 6 Arranging Tubes Symmetrically in a Fixed Angle Vertical Tube or JS 24 Series Swinging Bucket Rotor For example two three four or six tubes can be arranged symmetrically in a six place rotor To secure the rotor to the drive spindle hub during centrifugation J series rotors are equipped with devices that screw into the hub If the rotor is left in the centrifuge between runs tighten the tie down device before each run Some rotors are equipped with tie down assemblies These may be knobs that can be hand tightened when the rotor is installed and between runs if the rotor is left in the centrifuge Other tie down assemblies are tightened by turning the rotor lid knob J Series Rotors and Tubes Rotor Knob Rotor Tie down Screw Torque Bar OVERSPEED PROTECTION J Series Rotors and Tubes Rotors Some new and modified rotors have dual locking lid mechanisms The dual locking lid mechanism consists of a daisy knob that secures the lid to the rotor and a tie down knob that attaches the rotor to the centrifuge drive hub Daisy refers to the knob shape The grooves between each petal let your fingers grip the knob firmly and provide leverage for turning The daisy knob allows you to attach the lid to
132. ording to the instructions in your centrifuge instruction manual No addi tional input is required J Series Rotors and Tubes B 1 Temperature Compensation Tables Table B 1 Temperature Compensation Settings for the J2 HC Centrifuge Interpolate if intermediate values are required JA 25 50 10 000 5 3 2 18 000 8 E 45 JA 25 15 15 000 6 5 6 10 000 3 2 9 18 000 N 8 xg JA 21 15 000 6 6 N 10 000 4 29 N 18 000 N N 4 JA 20 1 15 000 7 5 2 10 000 4 D N 18 000 N N 3 JA 20 15 000 6 5 go 10 000 4 4 _9 16 000 N N 0 15 000 N N 0 SAIS 10 000 4 28 N 8 000 f D N 18 000 N N 4 17 000 N N 3 16 000 N N D dinis 12 000 N 6 8 000 3 2 N 5 000 1 N 17 000 N N 6 6 6 5 43 15 000 N 5 4 4 4 3 0 dele 12 000 4 3 3 3 N 8 000 20 2 p E aD a N 14 000 N N 7 T 7 5 4 JLA 16 250 10 000 N 4 3 3 9 zd N 5 000 N 0 0 3 3 3 N i Continued CAUTION For proper temperature control the JA 18 1 fixed angle rotor must be derated in the J2 HC Centrifuge as follows when the 25 degree angle adapters are used the maximum speed is 16 000 rpm when the 45 degree angle adapters are used the maximum speed is 15 000 rpm Above 30 C ambient temperature this temperature may not be achieved at this speed N indicates that the rotor cannot achieve the desired temperature at this speed B 2 J Series Rotors and Tubes Temperature Compensation Tables Table B 1
133. ormed by centrifugation the time it takes to form a gradient depends on the sedimentation and diffusion coefficients of the gradient material the pathlength and the rotor speed For a given gradient material the shorter the pathlength and the higher the rotor speed the faster the gradient will form In general the time required for gradients to reach equilibrium in swinging bucket rotors will be longer than in fixed angle rotors One way to reduce run times is to use partially filled tubes Refer to the applicable rotor manual to determine the maximum allowable speed and solution density when using partially filled tubes Particle separation achieved with rate zonal separation is a function of the particles sedimentation coefficient density size and shape and the viscosity of the gradient material Sucrose is especially useful as a gradient material for rate zonal separation because its physical characteristics are well known and it is readily available Samples are layered on top of the gradient Under centrifugal force particles migrate as zones Rate zonal separation is time dependent if the particles are more dense than the most dense portion of the gradient some or all of the particles will pellet unless the run is stopped at the appropriate time A separation is sometimes a combination of rate zonal and isopycnic Depending on particle buoyant densities and sedimentation coeffi cients some particles may be separated by their d
134. otor Preparation and Loading 0 000000 6 7 Operati n 2 212 bue bee br re RR Rd eed deg 6 8 Installing the Rotor leleee RR 6 9 Removal and Sample Recovery 0 00000 005 6 10 SECTION 7 CARE AND MAINTENANCE Rotor Care xiu er estes ae eas Pe Rb ata o adnata es 7 1 Cleaning iine XS chee ec batieh era tie es habia te 7 2 Decontamination 0 7 3 Sterilization and Disinfection 0 0 cece eee 7 4 INSPECHOM see cue sede cna Sp CERE E ee n Oe 7 4 Lubrication ssseeeeeeee n 7 5 Field Rotor Inspection Program 0 000000 0 eee 7 6 Tube Bottle and Accessory Care 00 e eee eens 7 7 Cleaning 2 2 22 ees does eg ee edd nee ed d 7 7 Decontamination 0 cece e 7 8 Sterilization and Disinfection 0 0 0 0 0c ee eee 7 8 Inspection es seed see eb eg RE eee d he 7 10 Tube and Bottle Storage 0 2 eee eee eee 7 10 Returning a Rotor or Accessory to the Factory 7 11 Diagnostic Hints 0 cece eens 7 12 Appendix A Chemical Resistances for Beckman Coulter Centrifugation Products A 1 Appendix B Temperature Compensation Tables B 1 Appendix C Gradient Materials C 1 Appendix D Blood Component Separation D 1 Appendix E Glossary of Terms E 1 Appendix F References 000 cece eee F 1 Warranty Vi
135. our fingers grip the knob firmly and provide leverage for turning The dual locking capability provides added biosafety by allowing the rotor to be loaded into and removed from the centrifuge with the lid in place The rotor may be placed under a safety hood before the lid is attached or removed A CAUTION Always loosen the rotor knob before loosening the daisy knob to avoid jamming the knobs A feature of many Beckman Coulter fixed angle rotors is a patented fluid containment annulus located below the O ring sealing surface If tubes are overfilled or if leakage occurs during centrifugation the annulus holds enough volume that all of the liquid is kept inside the rotor even if all tubes leak at the same time This feature virtually eliminates the escape of liquid into the centrifuge chamber I gt NOTE Although rotor components and accessories made by other manufacturers may fit in the Beckman Coulter rotor you are using their safety in the rotor cannot be ascertained by Beckman Coulter Use of other manufacturers components or accessories in the Beckman Coulter rotor may void the rotor warranty and should be prohibited by your laboratory safety officer Only the components and accessories listed in the applicable rotor manual should be used Fixed angle rotors can accommodate a variety of tube types listed in the rotor manual Refer to Section 3 USING TUBES AND BOTTLES for tube filling and sealing requirements Obser
136. particle sedimentation Using sedimentation theory users can calculate sedimentation rates For example red blood cells settle at the approx imate rate of 2 cm per hour in aqueous medium at 1 g with higher force fields increasing the settling rate Note that blood cells should not be subjected to high centrifugal force fields as the cells can be damaged D 4 J Series Rotors and Tubes Platelet rich Plasma Whole Blood Red Blood Cells Cell Free Packed Cells Blood Component Separation Plasma Platelet depleted Platelet Concentrate Plasma Cryoprecipitate Removed Cryoprecipitated Antihemophilic Factor Factor VIII Figure D 1 Blood Component Preparation Table D 2 Blood Bank Methods The speeds times and brake settings shown here are intended to be guidelines only Optimum conditions for separating blood components in each centrifuge must be determined by the user before carrying out actual separation runs Platelet Rich Whole Blood Soft Spin 20 to 22 JS 4 2 6 2800 3 0 6 1 4 5 5 5 Plasma and JS 4 2SM 6 2850 3 0 6or7 1 4 5 5 5 Red Blood Cells JS 5 2 4 3000 2 9 6 1 4 5 5 5 Platelet Platelet Rich Hard Spin 20 to 22 JS 4 2 6 3850 6 0 6 1 4 5 5 5 Concentrate Plasma JS 4 2SM 6 3900 6 0 6or7 1 4 5 5 5 JS 5 2 4 4100 6 0 6 1 4 5 5 5 Plasma Cell free Whole Blood Hard Spin 4 JS 4 2 6 3850 6 0 6 1 4 5 5 5 and Packed Cells JS 4 2SM
137. piercing the tube to avoid pushing the needle out the opposite side Puncture the bottom of the tube and collect the drops T Sample out p Aspirate the sample from the tube top by snipping off the tube stem then aspirating the sample with a Pasteur pipette or needle and syringe J Series Rotors and Tubes 3 21 Using Tubes Bottles and Accessories vH 4 CentriTube Slicer 347960 Slice the tube using the Beckman CentriTube Slicer 347960 Refer to publication L TB 010 for instructions for using the CentriTube Slicer For additional information on fraction recovery systems available from Beckman Coulter refer to the latest edition of High Perfor mance High Speed High Capacity Rotors Tubes amp Accessories publication BR 8102 MAKING ULTRA CLEAR TUBES WETTABLE 3 22 The following method of making Ultra Clear tubes wettable has proven successful for some users 1 Polyvinyl alcohol 2 g was dissolved in distilled water 50 mL by stirring and heating to gentle reflux Isopropanol 50 mL was slowly added to the hot solution and stirring and heating continued until a clear solution was obtained The solution was then allowed to cool to room temperature Ultra Clear tubes were filled with the coating solution then aspirated out with a water pump after 15 minutes leaving a thin film on the tube walls A small amount of solu
138. ply appropriate decontam ination procedures to the centrifuge rotor and accessories Turn the tie down knob to the left counterclockwise to release the rotor from the drive spindle 2 Lift the rotor straight up and off the drive spindle If the rotor sticks to the drive spindle a rotor removal tool may be used I gt NOTE Lubrication of the centrifuge drive spindle hub with Spinkote should prevent the rotor from sticking 3 To remove a tray from a carrier lift the end of the tray just enough to clear the carrier Slide the tray up so that it passes under the hinge pins J Series Rotors and Tubes ROTOR CARE J Series Rotors and Tubes 7 Care and Maintenance This section provides information on the care of rotors and accessories Included is a list of some common operating problems with suggestions for their solutions Rotors and accessories should be kept in optimal condition thus minimizing the chances of rotor or labware failure In addition to these instructions observe procedures and precautions provided in individual rotor manuals Appendix A of this manual provides the chemical resistances of rotor and accessory materials to various acids bases salts and solvents Rotor care involves not only careful operating procedures but also careful attention to Regular cleaning decontamination and or sterilization as required Frequent inspection Corrosion prevention and Regular and proper
139. r is a fixed angle rotor with a maximum speed of 12 000 rpm However the naming system for J series rotors was changed slightly in early 1994 Rotors released before 1994 for example the JA 18 1 JA designates that it is a fixed angle rotor used in a J series centrifuge the 18 indicates that the rated speed of the rotor is 18 000 rpm the decimal unit 1 distinguishes between different rotors with the same rated speed Rotors released after January 1994 for example the JA 25 50 JA still designates that it is a fixed angle rotor used in a J series centrifuge the 25 still identifies the rated speed of the rotor 25 000 rpm but the decimal unit 50 describes the nominal volume of the largest tube or bottle in mL used in the rotor An example of each rotor type is shown in Figure 1 1 1 1 Rotors Fixed Angle Rotor Swinging Bucket Rotor Vertical Tube Rotor Rack Type Rotor Figure 1 1 Fixed Angle Swinging Bucket Vertical Tube and Rack Type Rotors Containers in fixed angle rotors designated JA are held at an angle to the axis of rotation in tube cavities Containers in J Lite fixed angle rotors designated JLA are also held at an angle to the axis of rotation the rotor construction results in reduced overall weight Containers in swinging bucket rotors designated JS are held in rotor buckets or multitube carriers attached to the rotor body by hinge pins The buckets or carriers swing out to a hor
140. r with magnets without magnets maximum is 14 000 rpm Maximum speed at 2 C in a 50 Hz centrifuge is 14 000 rpm 4 Maximum speed in an Avanti J E for the rotor with magnets without magnets maximum is 13 000 rpm Maximum speed at 2 C in a 50 Hz centrifuge is 15 000 rpm Maximum speed in an Avanti J series centrifuge Maximum speed in a J2 series centrifuge is 14 000 rpm f Maximum speed in an Avanti J E for rotor without magnets is 6 300 rpm Maximum speed for rotor in an Avanti J E centrifuge is 6 300 rpm h The JS 24 38 and JS 24 15 rotors can achieve 24 000 rpm in an Avanti J 30 I centrifuge only In Avanti J 25 series and J 20 series centrifuges the maximum speed for these rotors is 10 000 rpm J Series Rotors and Tubes 1 7 Rotors 1 8 Ind Selection of a rotor depends on a variety of factors such as sample volume number of sample components to be separated particle size run time required quality of separation type of separation and the centrifuge in use Fixed angle swinging bucket vertical tube and rack type rotors are designed to provide optimal separations for a variety of sample types For especially large sample volumes contin uous flow and zonal rotors are available Fixed angle rotors are general purpose rotors that are especially useful for pelleting subcellular particles and in short column banding of viruses and subcellular organelles T
141. re held at an angle usually 20 to 45 degrees from the axis of rotation generic name for three leukocyte white blood cell types characterized by having granules in their cytoplasm centrifugation run 5 to 7 minutes at high g forces 4000 to 5000 x g at ambient temperature or at 4 C used to separate fresh plasma from cellular components high density polyethylene used for adapters a method of particle separation or isolation based on particle buoyant density particles are centrifuged until they reach a point in the gradient where the density of the particle is the same as the density of the gradient at that point low density polyethylene used for tubes and bottles procedure in which leukocytes or white blood cells are separated from whole blood see WBC white blood cells a type of leukocyte formed in the lymph nodes other lymphoid tissue and bone marrow about a quarter of the white blood cells in the circulating blood are lymphocytes the maximum volume at which a tube should be filled for centrifugation sometimes referred to as maximum fill volume or nominal fill volume the curved upper surface of a liquid column that is concave when the container walls are wetted by the liquid and convex when they are not sodium chloride a lower density salt than CsCl primarily used in lipopro tein type separations black synthetic elastomer used for O rings in some tube caps and bottle cap assemblies should be used at temper
142. rrier 368905 3 After centrifugation grasp the carrier by the wire handles and lift it straight up out of the bucket to unload it J Series Rotors and Tubes 5 15 Using Swinging Bucket Rotors J6 Carriers ECISMAWN watched Weight Microplate Carrier Run matched carriers 180 opposed on rotor Weight stamped on carrier frame Maximum Number Of Plates 3 C Use with only JS 5 2 and JS 4 0 rotors Le J OPERATION 5 16 Carriers used with the JS 5 2 and JS 4 0 rotors are NOT interchange able with those used with the JS 4 2 JS 4 2A JS 4 2SM JS 3 0 and JS 2 9 rotors If you have more than one type of carrier check the label on the side of the carrier to make sure that you are using the right one for your rotor Table 5 2 lists carriers used with J6 rotors 1 To prevent microtiter plate breakage during centrifugation place the rubber pad that comes with each carrier on the bottom of the carrier 2 Place the plate s on top of the pad being careful not to spill the contents Table 5 2 Microplate Carriers Used with J6 Series Rotors Maximum Rotor Number Rotor Run Carrier Set Type Carriers Load Speed Part Number JS 5 2 or 4 12 single 2600 rpm Set of 2 358680 JS 4 0 Set of 4 358681 JS 4 2 6 18 single or 2500 rom Set of 2 358682 JS 4 2A 6 deep well Set of 4 358683 JS 4 2SM Set of 6 358684 JS 3 0 or JS 2 9 Refer to the centrifuge instru
143. s horizontal position If it does remove the cup from the rotor and reposi tion the blood bag so that it seats further into the cup Allowing the blood bags to contact the rotor yoke during centrifugation can cause the bucket to come off the pivot pins and can seriously damage both the rotor and the centrifuge LOADING BUCKETS INTO THE ROTOR I gt NOTE JS 24 series rotors must have buckets attached before the rotor is put into the centrifuge 1 If bucket covers or rotor lids are used to help contain spills and Plug glass particles that could result from tube breakage make sure 949095 cover O rings are in good condition and lightly coated with silicone vacuum grease Before use inspect Aeroseal cover lt ee sealing surfaces especially the O ring groove It must be smooth and free of scratches Also ensure that the top 2 54 cm 1 in of O Ring the bucket is clean and smooth buckets with scratches or gouges 344660 in this surface will not seal properly Inspect the O ring and plug for nicks abrasions and other damage Replace damaged components Aeroseal Cover Assembly ee 2 Load the filled buckets and or carriers onto the rotor yoke pivot pins following the instructions in the rotor manual Make sure that the buckets are properly seated by gently swinging them on the pivot pins J Series Rotors and Tubes 5 13 Using Swinging Bucket Rotors 5 14 I gt NOTE All positions on the rotor yoke
144. s of bottle material and chemicals Bottles may crack or become brittle if they are used below their lower temperature limit Before using bottles at other than stated temperature limits evaluate them under centrifugation conditions If sample is frozen in bottles make sure that they are thawed to at least 2 C before centrifugation Bottles may become brittle with age and use Dispose of brittle or cracked bottles Improper cleaning decontamination or sterilization procedures used Refer to Table 7 1 for acceptable procedures and materials J Series Rotors and Tubes Appendix A Chemical Resistances for Beckman Coulter Centrifugation Products To Close Rotors and Tubes and Open the Chemical Resistances Chart Click Here J Series Rotors and Tubes A 1 Appendix B Temperature Compensation Tables This Appendix contains tables listing temperature compensation units for various rotors used in Beckman Coulter J series centrifuges TEMPERATURE COMPENSATION To ensure that the rotor reaches the required temperature during centrifugation some temperature compensation may be required because of the mass of these rotors The following tables list tempera ture compensation units for various rotors Refer to Section 1 or the applicable rotor manual for procedures to set the temperature compensation for the model of J centrifuge being used gt NOTE When using an Avanti J series centrifuge enter the run temperature acc
145. some vibration 5 4 J Series Rotors and Tubes Using Swinging Bucket Rotors LABWARE Swinging bucket rotors can accommodate a variety of tubes bottles multiwell titer plates and blood bags listed in individual rotor manuals Refer to Section 3 USING TUBES AND BOTTLES for tube filling and sealing requirements Observe the maximum rotor speeds and fill volumes listed in the applicable rotor manual ROTOR PREPARATION AND LOADING For runs at other than room temperature refrigerate or warm the rotor beforehand for fast equilibration PRERUN SAFETY CHECKS Read all safety information in the rotor manual before using the rotor 1 Make sure that the rotor and buckets or carriers are clean and show no signs of corrosion or cracking 2 Check the chemical compatibilities of all materials used Refer to Appendix A CHEMICAL RESISTANCES 3 Verify that the tubes bottles or carriers being used are listed in the applicable rotor manual 4 If fluid containment is required use capped tubes or bottles and or Aerosolve cannisters We strongly recommend capping all containers carrying physiological fluids to prevent leakage J Series Rotors and Tubes 5 5 Using Swinging Bucket Rotors ROTOR PREPARATION Lubrication Points between the pivot pin and rotor yoke all pivot pins Rotor Yoke Lubrication Points all pin surfaces and both pin sockets Be sure that metal threads in the rotor yoke are clean and l
146. such as immersion in 10 hydrogen peroxide for 30 minutes may be used on Ultra Clear tubes Refer to Table 7 1 to select cold sterilization materials that will not damage tubes and accessories 7 9 Care and Maintenance While Beckman Coulter has tested these methods and found that they do not damage the components no guarantee of sterility or disinfec tion is expressed or implied When sterilization or disinfection is a concern consult your laboratory safety officer regarding proper methods to use I gt NOTE INSPECTION Multiwell plates can be purchased already sterilized Inspect containers and accessories before use TUBE AND BOTTLE STORAGE Inspect tubes and bottles for cracks or any major deformities before using them Do not use a tube that has become yellowed or brittle with age or excess exposure to ultraviolet light Crazing the appearance of fine cracks on tubes and bottles is the result of stress relaxation If a crack approaches the outer wall of the tube or bottle discard it Discard any deformed or cracked adapters gt NOTE Replace the air vent filter in each JS 5 0 cup cover after every 250 cycles or after every auto clave cycle Tubes and bottles have an indefinite shelf life if properly stored Store in a dark cool dry place away from ozone chemical fumes and ultraviolet light sources 7 10 J Series Rotors and Tubes Care and Maintenance RETURNING A ROTOR OR ACCESSOR
147. sufficient to permit separation of the particles of interest by the chosen density gradient technique without overstressing the rotor e t should not affect the biological activity of the sample e t should be neither hyperosmotic or hypoosmotic when the sample is composed of sensitive organelles t should not interfere with the assay technique t should be removable from the purified product e t should not absorb in the ultraviolet or visible range e t should be inexpensive and readily available more expensive materials should be recoverable for reuse It should be sterilizable It should not be corrosive to the rotor t should not be flammable or toxic to the extent that its aerosols could be hazardous The following charts are provided as a reference for information on commonly used gradient materials Gradient Materials C 2 Table C 1 Commonly Used Gradient Materials with Their Solvents Sucrose 6696 Sucrose 6596 Silica sols Diodon Glycerol Cesium chloride Cesium formate Cesium acetate Rubidium chloride Rubidium formate Rubidium bromide Potassium acetate Potassium formate Sodium formate Lithium bromide Lithium chloride Albumin Sorbitol Ficoll Metrizamide J Series Rotors and Tubes Gradient Materials Table C 2 Density Refractive Index and Concentration Data Cesium Chloride at 25 C Molecular Weight 168 37
148. t the blood bags and tubing fit as far down as possible gt NOTE Make sure the bags stay as vertical as possible with no folds at the top or corners If folds are present blood cells could remain in the folds and then mix with the plasma when the bag is removed 2 Sandwich the tubing between the blood bag and any transfer packs see Figure 5 3 3 Make sure the loaded blood bag cups opposite each other on the rotor yoke are approximately the same weight within limits listed in the applicable rotor manual In some rotors balancing pads can be used if necessary to maintain weight balance Blood Bag Label Sandwich Coiled Tubing between Blood Bag Seam Ports Aligned with Transfer Bags Pivotal Axis of and Filled Bucket Blood Bag ES Blood Bag Seam Blood Bag Label Blood Bag Label Blood Bag Label Blo dBag Cup Figure 5 3 Typical Blood Bag Loading Procedure JS 4 3 Rotor Shown 5 12 J Series Rotors and Tubes Using Swinging Bucket Rotors 4 Place loaded cups into rotor buckets If only two filled cups are run place them in opposing buckets The remaining buckets should contain similar blank loads to prevent imbalance either empty modular disk adapters or water filled blood bags in cups A CAUTION If bucket covers or rotor lids are not used make sure the superstructure of the blood bag protruding from the cup does not inhibit the bucket from reaching it
149. talcum powder to the tube adapter centrifugation rubber bases after cleaning or as required to prevent sticking Tube leakage Tubes with cap assemblies Caps not properly secured Caps must be properly seated on tubes and then fully tightened Capcomponents not dry before assembly Thoroughly dry all components before assembling Tubes with snap on caps Tube too full the meniscus must be kept lower to prevent leakage Uncapped tubes Tube volume exceeds maximum uncapped volume Refer to the rotor manual for tube volumes and speed reductions Quick Seal tubes Improperly sealed After heat sealing squeeze the tube gently if the tube contents may be disturbed to test the seal for leaks If the tube leaks reseal it Tube cracking Tubes may crack or become brittle if they are used below their lower temperature limit Before using tubes at other than stated temperature limits evaluate them under centrifugation conditions If sample is frozen in tubes make sure that they are thawed to at least 2 C before centrifugation Tubes may become brittle with age and use Dispose of brittle or cracked tubes J Series Rotors and Tubes 7 13 Care and Maintenance SYMPTOM POSSIBLE CAUSE AND SUGGESTED ACTION Tube collapse Thinwall tube volume too low to provide tube wall support Meniscus should be 2 to 3 mm below the tube top Refer to the rotor manual for tube volumes Moisture between the tube and the cavity or bucket can cause
150. the contents 3 Slide the base pad and plate assembly into the carrier until the base clicks into place J Series Rotors and Tubes Using Swinging Bucket Rotors Deep well Plate Figure 5 4 The Micro Plus Microtiter Plate Carrier Base Pad and Deep Well Microtiter Plate JS 5 9 and JS 5 3 Plate Carriers High impact thermoplastic carriers are used in the JS 5 9 and JS 5 3 rotor buckets to provide support to labware during centrifugation and facilitate loading and unloading buckets Each rotor bucket can carry a 96 well kit for high throughput processing such as a DNA or RNA kit or standard microplates used in the serial dilution of small liquid volumes up to five JS 5 9 or six JS 5 3 stacked 96 well polypro pylene plates two stacked deep well plates or one square well plate per bucket JS 5 9 Carrier Carrier 368920 11 gt NOTE When using stacked polypropylene plates place a support pad beneath the bottom plate to prevent breakage during centrifugation Use the support pad beneath all polystyrene plates JS 5 3 Carrier support pad in the carrier with the ridged cross side down Support Pad 369382 1 If using polystyrene or stacked polypropylene plates place a S em J 2 Place the filled labware in the carrier If using more than one microplate per carrier place a cap strip between the plates to Z prevent breakage during centrifugation a
151. tion that collected in the tube bottoms after standing was removed with a pipette The tubes were left open to dry at room temperature overnight then filled with distilled water After standing overnight at room temperature the distilled water was poured out Finally the tubes were briefly flushed with water tapped to remove excess liquid and left to dry J Series Rotors and Tubes J N DESCRIPTION 4 Using Fixed Angle Rotors This section contains instructions for using fixed angle rotors in J series centrifuges In addition to these instructions observe procedures and precautions provided in the applicable rotor and centrifuge manuals Refer to Section 2 for labware selection information and Section 3 for recommended filling and sealing or capping requirements and for sample recovery procedures Refer to Section 7 for information on the care of rotors and accessories Fixed angle rotors see Figure 4 1 are general purpose rotors that are especially useful for pelleting subcellular particles and in short column banding of viruses and subcellular organelles Refer to Table 4 1 for general rotor specifications Tubes in fixed angle rotors are held at an angle usually 20 to 45 degrees to the axis of rotation The tube angle shortens the particle pathlength compared to swinging bucket rotors resulting in reduced run times Tubes can be placed directly in a rotor cavity if the diameters of the tube and th
152. tor in which tubes are placed in gamma counter racks the racks are loaded into special plastic trays which are then loaded into carriers that swing up to the horizontal position during centrifugation polyphenylsulfone PPS used in plugs cap closures cannisters and other accessories a method of particle separation based on differential rate of sedimentation using a preformed gradient with the sample layered as a zone on top of the gradient red blood cells or erythrocytes carry oxygen to the tissues and carbon dioxide to the lungs for exhalation J Series Rotors and Tubes RCF Tmax Tmin SAG M sedimentation sedimentation coefficient s SDS silicone rubber soft spin Solution 555 J Series Rotors and Tubes Glossary of Terms relative centrifugal field the ratio of the centrifugal acceleration at a specified radius and speed r to the standard acceleration of gravity g according to the following equation RCF ro E where r is the radius in millimeters is the angular velocity in radians per second 27 RPM 60 and g is the standard acceleration of gravity 9807 mm s2 Thus the relationship between RCF and RPM is 2 en maximum radius the position of the liquid in the tube at the maximum distance from the axis of rotation when the rotor is at speed minimum radius the position of the liquid in the tube at the minimum distance from the axis of rotation when the rotor is at speed
153. ube to deform disrupting gradients or sample Spacer and or floating spacer require ments for Quick Seal tubes are described in the individual rotor manuals The neck of the tube should be clean and dry before sealing 3 A sample application block part number 342694 is available for holding and compressing tubes and can be used to layer samples on preformed gradients in polyallomer Quick Seal tubes J Series Rotors and Tubes 3 9 Using Tubes Bottles and Accessories There are two tube sealers for use with Quick Seal tubes the hand held Cordless Tube Topper and the older tabletop model no longer available Refer to How to Use Quick Seal9 Tubes with the Beckman Coulter Cordless Tube Topper M publication IN 181 for detailed information about the Tube Topper Instructions for using the older tabletop tube sealer are in How to Use Quick Seal9 Tubes with the Beckman Tube Sealer publication IN 163 Quick Seal tubes are heat sealed quickly and easily using the Beckman Coulter Cordless Tube Topper see Figure 3 1 The following procedures provide the two methods for heat sealing Quick Seal tubes using the hand held Tube Topper Use the applicable tube rack listed in the rotor manual Charging Stand Figure 3 1 The Cordless Quick Seal Tube Topper N CAUTION Before plugging in the Tube Topper be sure that you have a proper power source 120 V 50 or 60 Hz Charge your Cordless Tube Topper only in the charging stand supp
154. ubes are held at an angle usually 20 to 45 degrees to the axis of rotation The tube angle shortens the particle pathlength see Figure 1 2 compared to swinging bucket rotors resulting in reduced run times Tubes can be placed directly in a rotor cavity if the diameters of the tube and the cavity are the same Using adapters more than one type and size of tube can be centrifuged together provided that the loads are properly balanced Refer to Section 4 for specific infor mation about the use of fixed angle rotors Swinging bucket rotors are used for pelleting isopycnic studies separation as a function of density and rate zonal studies sepa ration as a function of sedimentation coefficient Large swinging bucket rotors are used to obtain cell free plasma or for cell packing These rotors can be equipped with racks or microplate carriers to hold a variety of tubes bottles blood bags or multiwell plates Refer to Section 5 for specific information about the use of swinging bucket rotors Vertical tube rotors hold tubes parallel to the axis of rotation therefore bands separate across the diameter of the tube rather than down the length of the tube see Figure 1 2 Only Quick Seal tubes are used in vertical tube rotors making tube caps unneces sary Refer to Section 6 for specific information about the use of vertical tube rotors Rack type rotors hold tubes in gamma counter racks Racks are loaded into special plastic
155. urs amp Company J Series Rotors and Tubes Rotors ROTOR SELECTION Rotors used in Beckman Coulter J series centrifuges are listed in Table 1 1 General rotor specifications for each fixed angle rotor are in Table 4 1 swinging buckets in Table 5 1 and vertical tube and rack type in Table 6 1 Detailed descriptions of each rotor are included in the applicable rotor manual Table 1 1 Rotors Used in Beckman Coulter J Series Centrifuges Rotors in parentheses are no longer manufactured Fixed Angle JA 30 50 Ti 400 mL 30 000 108 860 280 X X X X X X JA 25 50 400 mL 25 000 76 6008 418 X X X X X X X JA 25 15 360 mL 25 000 74 200 265 X X X X X X outer row 60 200 380 inner row JA 21 180 mL 21 000 50 400 470 X X X X X X X X X X JA 20 1 480 mL 20 000 51 500 325 X X X X X X X X X X outer row 43 900 371 inner row JA 20 400 mL 20 000 48 400 770 X X X X X X X X X X JA 18 1 43 2 mL 18 000 42 100 156 X X X X X X X X X 45 angle adapter JA 18 1 43 2 mL 17 000 36 300 91 X X X X X X X X X 25 angle adapter Continued a Maximum speed in an Avanti J E centrifuge is 21 000 rpm 18 000 rpm at 2 C at 35 C ambient and 95 percent humidity b When a JA 18 1 rotor is used in a J2 HC centrifuge derate the rotor as follows when the 45 adapters are used do not run the rotor above 15 000 rpm when 25 adapters are used do not run the rotor above 16 000 rpm
156. used are in good condition and are listed in the rotor manual J Series Rotors and Tubes 6 7 Using Vertical Tube and Rack Type Rotors Rotor Preparation Attachment OPERATION 6 8 1 Lubricate the rotor drive hole with silicone vacuum grease 335148 2 Load racks into either two or four trays then load tubes into the racks Do not over fill tubes leave enough space to avoid spills during carrier loading I gt NOTE Fill all tubes to the same level with liquid of the same density Racks and tubes must be horizon tally and vertically symmetrical during centrifugation 3 Rest the end of the loaded tray on the carrier base Slide the tray down so that it passes under the hinge pins When it reaches the lower end of the carrier seat the tray bottom completely into the catrier Refer to the centrifuge instruction manual for detailed operating information For low temperature runs precool the rotor in the centri fuge or in a refrigerator before use especially before short runs to ensure that the rotor reaches the set temperature To ensure that the rotor reaches the required temperature during centrifugation some temperature compensation may be required because of the mass of these rotors Refer to Appendix B or to the rotor manual for tables listing temperature compensation units for various rotors If you are using a microprocessor controlled J6 series centrifuge enter the rotor code enter code 3 2 for t
157. utoclaved many times or if they are handled or compressed before they cool Thinwall polyallomer tubes should be placed open end down or supported in a rack if autoclaved N CAUTION Do not autoclave tubes or bottles with caps on Pressure in a sealed container can cause an explosion Pressures within the autoclave can cause partially sealed containers to collapse when the autoclave vents JS 5 0 labware cups cup covers cup gaskets 369261 and 369257 and partitions can be autoclaved at 121 C for up to 20 minutes Remove the plug and air vent filter from each cup cover before autoclaving and remove the gasket from the cup To remove an air vent filter gently push it out from underneath the cover with a pencil or other non metal tool that will not scratch the cover material After autoclaving insert a new air vent filter into each cup cover Thoroughly dry the gasket sealing surfaces before replacing the gasket N CAUTION Autoclaving will reduce the useful life of the labware cups cup covers cup gaskets and partitions After each autoclave cycle examine these components for damage and DO NOT USE damaged components HarvestLine system liners can be gamma irradiated to a maximum dose of 40 0 kGy Gamma irradiation causes the liners to become yellow but does not affect their performance Do not steam or dry autoclave the liners or they will be damaged The liners are designed for single use only A cold sterilization method
158. ve the maximum rotor speeds and fill volumes listed in the rotor manual J Series Rotors and Tubes Using Fixed Angle Rotors IB NOTE JLA 8 1000 and JLA 9 1000 rotors run only the specially designed bottles with polycarbonate seals and Radel cap closures Refer to the appli cable rotor manual for instructions on use of these bottles and accessories ROTOR PREPARATION AND LOADING PRERUN SAFETY CHECKS ROTOR PREPARATION J Series Rotors and Tubes For runs at other than room temperature refrigerate or warm the rotor beforehand for fast equilibration Read all safety information in the rotor manual before using the rotor 1 Make sure that the rotor and lid are clean and show no signs of corrosion or cracking Check the chemical compatibilities of all materials used Refer to Appendix A CHEMICAL RESISTANCES Verify that the tubes and bottles being used are listed in the appli cable rotor manual If fluid containment is required use capped tubes or bottles We strongly recommend capping all containers carrying physiological fluids to prevent leakage Be sure that metal threads in the rotor are clean and lightly but evenly lubricated with Spinkote lubricant 306812 Also ensure that O rings are lightly but evenly coated with silicone vacuum grease 335148 Using Fixed Angle Rotors 4 8 OPERATION Rotor JA 25 50 INSTALLING THE ROTOR 2 Load the filled containers symmetrically
159. without magnets is 13 000 rpm Maximum speed in a J2 series centrifuge is 14 000 rpm Maximum speed in an Avanti J E for a rotor with magnets maximum speed for a rotor without magnets is 13 000 rpm g Maximum speed in an Avanti J E for the rotor with magnets without magnets maximum is 14 000 rpm Maximum speed at 2 C in a 50 Hz centrifuge is 14 000 rpm Do not put bottles directly into the rotor without cannisters J Series Rotors and Tubes Using Fixed Angle Rotors Table 4 1 General Specifications for Beckman Coulter J Series Fixed Angle Rotors continued JA 14 14 000 rpm 600 to 137 86 35 6 x 250 mL 1 5 liter 25 Angle 30 100x g 800 JA 12 12000 rpm 400 to 144 108 71 12x50 mL 600 mL 35 Angle 23 200 x g 1000 1 244 JA 10 10000 rpm 600 to 158 98 38 6 x 500 mL 3 liters 25 Angle 17 700 x g 800 3610 JLA 10 500e 10 000 rom 600 to 166 110 64 6 x 500 mL 3 liters 20 Angle 18 600 x g 800 2 850 JLA 9 1000h 9 000 rpm 200 to 185 134 82 4 x 1000 mL 4 liters 20 Angle 16 800 x g 400 use only in 2 540 Avanti J series centrifuges JLA 8 1000h 8000rpm 200to 222 171 119 6 x 500 mL 6 liters f 20 Angle 15 900 x g 400 use only in 2 500 Avanti J 20 series and Avanti J HC centrifuges a Maximum speeds are based on a solution density of 1 2 g mL in all rotors except for the JA 18 1 which is rated for a density of 1 4 g mL b Critical speed rang
160. y information printed on the original solution containers prior to their use Handle body fluids with care because they can transmit disease No known test offers complete assurance that they are free of micro organisms Some of the most virulent Hepatitis B and C and HIV I V viruses atypical mycobacteria and certain systemic fungi further emphasize the need for aerosol protection Handle other infec tious samples according to good laboratory procedures and methods to prevent spread of disease Because spills may generate aerosols observe proper safety precautions for aerosol containment Do not run toxic pathogenic or radioactive materials in the rotor without taking appropriate safety precautions Biosafe containment should be used when Risk Group II materials as identified in the World Health Organization Labora tory Biosafety Manual are handled materials of a higher group require more than one level of protection Dispose of all waste solutions according to appropriate environmental health and safety guidelines e fdisassembly reveals evidence of leakage you should assume that some fluid escaped the container or rotor Apply appropriate decontamination procedures to the centrifuge rotor and accessories J Series Rotors and Tubes Mechanical Safety Useonly the rotors components and accessories designed for use in the rotor and centrifuge being used refer to the applicable rotor manual The safety of rotor c

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