Type 50.2 Ti Rotor
Contents
1. 8 up cos where dgis the interband distance when the tube is held at an angle in the rotor 17 Type 50 2 Ti Rotor 1 9 full 3 411 2 1 I oF 1 7 1 6 te a 1 5 aT SS Homogeneous CsCl Solution g mL TYPE 50 2 Ti ROTOR 13 20 C PP aE c i The dashed lines are a representation of See equation 6 and are drawn here to show the inability of that formula to guard against CsCl precipitation Note that at the lower right the precipitation curves intersect with 1 2 the square root deration lines in this case the lower speeds of the equation 6 prevail A 0 10 20 30 40 50 Rotor Speed K rpm Figure 2 Precipitation Curves for the Type 50 2 Ti Rotor Using combinations of rotor speeds and homogeneous CsCl solution densities that intersect on or below these curves ensures that CsCl will not precipitate during centrifugation Fill volumes are indicated on the curves The2. eee eee eee ee 468 mL Approximate acceleration time to maximum speed fully loaded 20 cece ee eee 10 1 2 min Approximate deceleration time from maximum speed fully loaded 02 eee 10 min Weight of fully loaded rotor 0 000005 13 6 kg 30 1b Rotor material o 4 ss ha sess sleet ckew bak ose bape eect ge cba soe ee titanium Relative Centrifugal Field RCF is the ratio of the centrifugal acceleration at a specified radius and speed r to the standard acceleration of gravity g according to the following formula RCF 12 E where r is the radius in millimeters is the angular velocity in radians per second 2 7 RPM 60 and g is the standard acceleration of gravity 9807 mm s After substitution RPM RCF 1 12r a Type 50 2 Ti Rotor DESCRIPTION Rotor Handle oo O ring large Lid O ring 834301 small 854519 i Overspeed e m Disk 830336 Rotor Body This rotor has been manufactured in a registered ISO 9001 or 13485 facility for use with the appropriately classified Beckman Coulter ultracentrifuges The Type 50 2 Ti rated for 50 000 rpm is a fixed angle rotor designed to centrifuge up to twelve tubes at a 24 degree angle to the axis of rotation Used in Beckman Coulter class H R and S preparative ultracentrifuges the rotor develops centrifugal forces sufficient for the differential separation of particles
3. 50 2 Ti rotor Curves are provided at two temperatures 20 C gray curves and 4 C black curves The reference curves in Figure 3 show gradient distribution at equi librium Each curve in Figure 3 is within the density limits allowed for the Type 50 2 Ti rotor each curve was generated for a single run speed using the maximum allowable homogeneous CsCl densities one for each fill level that avoid precipitation at that speed The gradients in Figure 3 can be generated from step or linear gradients or from homogeneous solutions But the total amount of CsCl in solu tion must be equivalent to a homogeneous solution corresponding to the concentrations specified in Figure 2 Figure 3 can also be used to approximate the banding positions of sample particles Figures 2 and 3 show that several fill volumes are possible in a tube If a tube is partially filled with gradient solution float mineral oil or some other low density immiscible liquid on top of the tube contents to fill the tube to its maximum volume Do not use an oil overlay in Ultra Clear tubes Note that for a given CsCl density as the fill level decreases the maximum allowable speed increases Partial filling may be desirable when there is little sample or when you wish to shorten the run time Type 50 2 Ti Rotor For example a quarter filled tube of 1 615 g mL homogeneous CsCl solution at 4 C may be centrifuged at 43 500 rpm see Figure 2 The same solution in a half
4. Failure to keep these threads lubricated can result in damaged threads e Regularly apply silicone vacuum grease 335148 to the O rings Replace O rings about twice a year or whenever worn or damaged Centering Tool 331325 CLEANING Kit 339558 Rotor Handle 337904 e O ring large pae 834301 Lid O ring A Rotor Cleaning small 854519 Type 50 2 Ti Rotor e Regularly inspect the overspeed disk If it is scratched damaged or missing replace it Rotors and Tubes contains instructions for replacing the overspeed disk Refer to the chemical resistances table in Appendix A of Rotors and Tubes for chemical compatibilities of rotor and accessory materials Your Beckman Coulter representative provides contact with the Field Rotor Inspection Program and the rotor repair center Wash the rotor 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 Under normal use wash the rotor frequently at least weekly to prevent buildup of residues 1 Remove the O rings before washing 2 Wash the rotor and lid in a mild detergent such as Beckman Solution 555 339555 that won t damage the rotor The Rotor Cleaning Kit 339558 contains two special plastic coated brushes and two quarts of Solution 555 for use with rotors and accessories Dilute the detergent 10 to 1 with
5. Ltd Oakley Court Kingsmead Business Park London Road High Wycombe Buckinghamshire HP11 1JU England U K Telephone 44 01494 441181 Fax 44 01494 447558 e mail beckman_uk beckman com Additional addresses are available at www beckmancoulter com 4300 N Harbor Boulevard Box 3100 Fullerton California 92834 3100 Internet www beckmancoulter com Telex 678413 Fax 1 800 643 4366 2007 Beckman Coulter Inc G Bd Printed on recycled paper
6. and for equilib rium sedimentation of particles such as DNA and viruses Up to 468 mL of gradient and sample volume can be centrifuged per run The rotor is made of titanium and is finished with black polyurethane paint The aluminum lid and handle are anodized for corrosion resis tance O rings made of Buna N rubber in the lid and handle maintain atmospheric pressure inside the rotor during centrifugation if they are properly lubricated Four small holes in the lid provide a temporary vent preventing pressure build up and extrusion of the large lid O ring in the event of tube leakage Because of the weight of the rotor drive pins are not required in the rotor drive hub cavity For overspeed protection a photoelectrical detector in the ultracentri fuge monitors the overspeed disk on the rotor bottom and shuts down the run if speeds exceeding 50 000 rpm are detected See the Warranty at the back of this manual for warranty information PREPARATION AND USE Specific information about the Type 50 2 Ti rotor is given here Infor mation common to this and other rotors is contained in Rotors and Tubes for Preparative Ultracentrifuges publication LR IM which should be used together with this manual for complete rotor and accessory operation PRERUN SAFETY CHECKS oll a Yo fe ear Be 330336 o gt anv ROTOR PREPARATION Type 50 2 Ti Rotor I gt NOTE Although rotor components and accessories made by other manufa
7. dashed lines represent equation 6 and are shown here to illustrate the inability of that equation to guard against CsCl precipitation 18 Type 50 2 Ti Rotor 1 90 1 80 f 1 70 1 60 ee VY Sa 1 50 rO a D ak P E D a oO a 1 40 8 1 30 AAS 7 Roa Sey DA lt lt X 1 20 FAN x TYPE 50 2 Ti ROTOR Tar 20 C PS AN 4 C b Each square on the 1 10 grid equals 0 010 g mL by 1 27 mm ia 00 009 D PT 0 000 z z 1 00 54 4 67 8 81 2 94 6 107 9 min Distance from Axis of Rotation mm max Figure 3 CsCl Gradients at Equilibrium for the Type 50 2 Ti Rotor Centrifugation of homogeneous CsCl solutions at the maximum allowable speeds from Figure 2 results in gradients presented here 19 Type 50 2 Ti Rotor Example B Knowing particle buoyant densities 1 60 and 1 65 g mL how do you achieve good separation 1 In Figure 3 sketch in a horizontal line corresponding to each pa
8. disinfec tion is a concern consult your laboratory safety officer regarding proper methods to use Refer to publication IN 192 included in each box of tubes or bottles for tube and bottle sterilization and disinfection procedures Quick Seal and thinwall open top tubes are disposable and should be discarded after a single use STORAGE When the rotor is not in use store it in a dry environment not in the instrument with the lid removed to allow air circulation so moisture will not collect in the tube cavities 3 Flammability hazard Do not use in or near operating centrifuges 23 Type 50 2 Ti Rotor 24 RETURNING A ROTOR 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 Beckman Coulter sales office and should contain the following information e serial number e history of use approximate frequency of use e reason for the return e original purchase order number billing number and shipping number if possible e name and phone number of the person to be notified upon receipt of the rotor or accessory at the factory and e 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 all parts are free
9. fill volumes 16 5 mL when centrifuged without cap maximum speed is 20 000 rpm ENoryl is a registered trademark of GE Plastics P Includes the 344672 neoprene gasket and 346246 stem 1 Tube cap is optional Type 50 2 Ti Rotor Table 1 Available Tubes and Bottles for the Type 50 2 Ti Rotor continued Tube Required Accessory Max Speed Part Max Part RCF Dimensions Description Number Fill Volume Description Number k Factor 13x 64mm stainless steel 301099 6 5 mL Delrin adapter 303392 30 000 rpm 94 700 x g aluminum cap 346256 126 or 305022 13x 64mm_ thickwall 355644 4mL Delrin adapter 303392 43 000 rpm polyallomer 194 000 x g 61 13x 64mm_ thickwall 355645 4mL Delrin adapter 303392 43 000 rom polycarbonate 194 000 x g 61 4 Refer to the section on RUN SPEEDS before using stainless steel tubes b Delrin is a registered trademark of E I Du Pont de Nemours amp Company Includes disposable plastic plugs dUltem is a registered trademark of GE Plastics For lower fill volumes 16 5 mL when centrifuged without cap maximum speed is 45 000 rpm f For lower fill volumes 16 5 mL when centrifuged without cap maximum speed is 20 000 rpm amp Nory is a registered trademark of GE Plastics P Includes the 344672 neoprene gasket and 346246 stem i Tube cap is optional Guus gt 25 C a Pl a m Stem Base A Meniscus of Stem r Tube Te
10. from pathogens and or radioactivity Steriliza tion and decontamination must be done before returning the parts Smaller items such as tubes bottles etc should be enclosed in a sealed plastic bag All parts must be accompanied by a note plainly visible on the out side of the box or bag stating that they are safe to handle and that they are not contaminated with pathogens 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 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 SUPPLY LIST Type 50 2 Ti Rotor I gt NOTE Publications referenced in this manual can be obtained by calling Beckman Coulter at 1 800 742 2345 in the United States or by contacting your local Beckman Coulter office See the Beckman Coulter Ultracentrifuge Rotors Tubes amp Accesso ries Catalog BR 8101 available at www beckmancoulter com for detailed information on ordering parts and supplies or contact Beckman Coulter sales 1 800 742 2345 in the United States world wide offices are listed on the back cover of this manual For your convenience a partial list is given below REPLACEMENT ROTOR PARTS OTHER Type 50 2 Ti roto
11. protection Handle other infectious samples according to good laboratory procedures and methods to prevent spread of disease Because spills may generate aerosols observe proper safety precautions for aerosol contain ment Do not run toxic pathogenic or radioactive materials in this centrifuge 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 The rotor and accessories are not designed for use with materials capable of developing flammable or explosive vapors Do not centrifuge such materials in nor handle or store them near the ultracentrifuge Although rotor components and accessories made by other manufacturers may fit in the Type 50 2 Ti rotor their safety in this rotor cannot be ascertained by Beckman Coulter Use of other manufacturers components or accessories in the Type 50 2 Ti rotor may void the rotor warranty and should be prohibited by your laboratory safety officer Only the compo nents and accessories listed in this publication should be used in this rotor Make sure that filled containers are loaded symmetrically into the rotor and that opposing tubes are filled to the same level with liquid of the same density Make sure that cavities in use have the proper spacers inserted if applicable before installing th
12. rpm bell top pkg 56 pkg 2 302 000 x g 68 25x 89mm_ thickwall 355631 30 mL w cap blue anodized 338906 50 000 rpm polycarbonate 16 5 mL cap 302 000 x g without cap 69 25x 89mm thickwall 355642 30 mL w cap blue anodized 338906 50 000 rpm polyallomer 16 5 mL cap 302 000 x g without cap 69 25x 89mm polycarbonate 355618 26 3 mL max aluminum cap 355619 50 000 rpm bottle assembly bottle only 16 mL min 302 000 x g 355654 68 4 Refer to the section on RUN SPEEDS before using stainless steel tubes continued bDelrin is a registered trademark of E I Du Pont de Nemours amp Company Includes disposable plastic plugs dUltem is a registered trademark of GE Plastics For lower fill volumes 16 5 mL when centrifuged without cap maximum speed is 45 000 rpm f For lower fill volumes 16 5 mL when centrifuged without cap maximum speed is 20 000 rpm Nory is a registered trademark of GE Plastics P Includes the 344672 neoprene gasket and 346246 stem i Tube cap is optional Table 1 Available Tubes and Bottles for the Type 50 2 Ti Rotor continued Type 50 2 Ti Rotor Tube Required Accessory Max Speed Part Max Part RCF Dimensions Description Number Fill Volume Description Number k Factor 25 x 64mm Quick Seal 344323 27 mL Noryl amp floating 343448 50 000 rom Ultra Clear spacer 302 000 x g 55 25 x 64mm Quic
13. the tube Insert Vise tube fi uoe ron 305075 cap vise 305075 e Tighten bottle caps by hand Polycarbonate Bottles The polycarbonate bottles may be centrifuged completely filled or partially filled not less than half full Again all opposing containers for a run must be filled to the same level Be sure to note the reduc tions in run speed shown in Table 1 if bottles are partially filled RUN TIMES TIME HR MIN The k factor of the rotor is a measure of the rotor s pelleting effi ciency Beckman Coulter has calculated the k factors for all of its He ee preparative rotors at maximum rated speed and using full tubes The k factor is calculated from the formula 7 In nax min 10 x 1 pe 3600 where is the angular velocity of the rotor in radians per second 0 105 x rpm rmax is the maximum radius and rmin is the minimum radius 12 RUN SPEEDS SPEED RPM RCF can oer pa N D V yu Type 50 2 Ti Rotor After substitution 2 533 x 10 n r nax Fmnin rpm 2 Use the k factor in the following equation to estimate the run time t in hours required to pellet particles of known sedimentation coeffi cient s in Svedberg units Run times can be estimated for centrifugation at less than maximum speed by adjusting the k factor as follows 2 E 50000__ Kaaj a run speed 4 Run times can also be estimated from data established in prior experi ments using a d
14. warranty covers the Beckman Coulter Centrifuge Systems only including but not limited to the centrifuge rotor and accessories and Beckman Coulter shall not be liable for damage to or loss of the user s sample non Beckman Coulter tubes adapters or other rotor contents 4 This warranty is void if the Beckman Coulter Centrifuge Sys tem is determined by Beckman Coulter to have been operated or maintained in a manner contrary to the instructions in the operator s manual s for the Beckman Coulter Centrifuge System components in use This includes but is not limited to operator misuse abuse or negligence regarding indicated main tenance procedures centrifuge and rotor classification require ments proper speed reduction for the high density of certain fluids tubes and tube caps speed reduction for precipitating gradient materials and speed reduction for high temperature operation 5 Rotor bucket sets purchased concurrently with or subsequent to the purchase of a Swinging Bucket Rotor are warranted only for a term co extensive with that of the rotor for which the bucket sets are purchased 6 This warranty does not cover the failure of a Beckman Coulter rotor in a centrifuge not of Beckman Coulter manufacture or if the rotor is used in a Beckman Coulter centrifuge that has been modified without the written permission of Beckman Coulter or is used with carriers buckets belts or other devices not of Beckman Coulter manufactur
15. water lII NOTE Do not wash rotor components in a dishwasher Do not soak in detergent solution for long periods such as overnight 3 Thoroughly rinse the cleaned rotor and components with distilled water 4 Air dry the rotor and lid upside down Do not use acetone to dry the rotor 5 Apply a thin even coat of silicone vacuum grease to the lid O ring before replacing it in the groove in the outer rim of the lid Also apply silicone vacuum grease to the handle O ring before reassembly 21 Type 50 2 Ti Rotor DECONTAMINATION Clean metal threads every 6 months or as necessary Use a brush and concentrated Solution 555 Rinse and dry thoroughly then lubricate lightly but evenly with Spinkote to coat all threads Periodically remove the O rings and wipe clean as necessary Clean the O ring grooves with a cotton tipped swab Reapply a light film of silicone vacuum grease If the rotor and or accessories becomes contaminated with radioac tive material it should be decontaminated 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 gt NOTE IsoClean can cause fading of colored anodized surfaces Use it only when necessary and remove it promptly from surfaces While Beckman Coulter has tested these methods and found that they do not
16. 0 NETHERLANDS Beckman Coulter Nederland B V Nijverheidsweg 21 3641 RP Mijdrecht Postbus 47 3640 AA Mijdrecht The Netherlands Telephone 31 297 230630 Fax 31 297 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 Espafia S A C Caleruega 81 28033 Madrid Spain Telephone 34 91 3836080 Fax 34 91 3836096 email bioresearch espana beckman com SWEDEN Beckman Coulter AB Archimedesvaegen 7 Box 111 56 SE 168 11 Bromma Sweden Telephone 46 8 564 85 900 Telefax 46 8 564 85 901 SWITZERLAND Beckman Coulter International S A 22 Rue Juste Olivier Case Postale 301 303 CH 1260 Nyon Switzerland Telephone 41 0800 850 810 Fax 41 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
17. COULTER 2007 Beckman Coulter Inc Printed in the U S A L5 TB 029P December 2007 Type 50 2 Ti Rotor Used In Beckman Coulter Class H R and S Preparative Ultracentrifuges Type 50 2 Ti Rotor SAFETY NOTICE This safety notice summarizes information basic to the safe use of the rotor described in this manual The international symbol displayed above is a reminder to the user that all safety instructions should be read and understood before operation or maintenance of this equipment is attempted When you see the symbol on other pages throughout this publication pay special attention to the specific safety information presented Observance of safety precautions will also help to avoid actions that could damage or adversely affect the performance of the rotor This rotor was developed manufactured and tested for safety and reliability as part of a Beckman Coulter ultracentrifuge rotor system Its safety or reliability cannot be assured if used in a centrifuge not of Beckman Coulter s manufacture or in a Beckman Coulter ultracentrifuge that has been modified without Beckman Coulter s approval A E gt ei ie ea Handle body fluids with care because they can transmit disease No known test offers complete assurance that such fluids are free of micro organisms Some of the most virulent Hepatitis B and C viruses HIV I V atypical mycobacteria and certain systemic fungi further emphasize the need for aerosol
18. Case Postale 301 303 CH 1260 Nyon Switzerland Telephone 41 22 365 3707 Fax 41 22 365 0700 FRANCE Beckman Coulter France S A Paris Nord II 33 66 rue des Vanesses B P 50359 Villepinte France 95942 ROISSY CDG Cedex Telephone 33 1 49 90 90 00 Fax 33 1 49 90 90 10 e mail bioresearch france beckman com GERMANY Beckman Coulter GmbH Europark Fichtenhain B 13 47807 Krefeld Germany Telephone 49 21 51 33 35 Fax 49 21 51 33 33 e mail bioresearch de beckman com GS zraou COULTER Beckman Coulter Inc e Sales and Service 1 800 742 2345 HONG KONG Beckman Coulter Hong Kong Ltd 12th Floor Oxford House 979 King s Road Taikoo Place Hong Kong Telephone 852 2814 7431 Fax 852 2873 4511 INDIA Beckman Coulter India Pvt Ltd Solitaire Corporate Park 3rd Floor Bldg 11 Andheri Ghatkopar Link Road Chakala Andheri West Mumbai India 400 098 Telephone 91 22 3080 5101 ITALY Beckman Coulter S p a Centro Direzionale Lombardo Palazzo F 1 Via Roma 108 20060 Cassina de Pecchi Milano Italy Telephone 39 02 953921 Fax 39 02 95392264 JAPAN Beckman Coulter K K TOC Ariake West Tower 2 5 7 Ariake Koto ku Tokyo 135 0063 Japan Telephone 81 3 5530 8500 Fax 81 3 5404 8436 MEXICO Beckman Coulter de Mexico S A de C V Avenida Popocat petl 396 Colonia Gral Pedro Maria Anaya Codigo Postal 03340 Mexico D F Mexico Telephone 001 52 55 9183 280
19. ailed information on the use and care of Quick Seal tubes Thinwall Tubes Thinwall polyallomer and Ultra Clear tubes require caps for tube support Fill the tubes as full as possible to prevent tube collapse during centrifugation If necessary float mineral oil or some other low density immiscible liquid on top of the tube contents to fill the tube to its maximum volume Do not use an oil overlay in Ultra Clear tubes Thickwall Tubes Thickwall polyallomer and polycarbonate tubes can be run partially filled at least half filled with or without caps but all opposing tubes for a run must be filled to the same level with liquid of the same density Do not overfill capless tubes be sure to note the reductions in fill volume and run speed shown in Table 1 11 Type 50 2 Ti Rotor Tube Caps Thinwall tubes require caps for tube support during centrifugation thickwall tubes and bottles can be run with or without caps speed reduction may be required Use only the caps listed in Table 1 Refer to Rotors and Tubes for the use assembly and maintenance of caps e Inspect tube caps before use as described in Rotors and Tubes Replace any damaged components e Inspect the O rings or gaskets in the caps for cracks nicks or flattened areas Be sure that they are dry and free of lubricant during assembly e Tighten red and blue anodized aluminum caps with a torque rsa wrench to 11 NeM 100 in Ib while the tube is held in
20. bes 361668 Spinkote lubricant 2 0Z 0 eee eens 306812 Silicone vacuum grease 1 OZ 0 eee eee eee eee 335148 Rotor Cleaning Kit 00 0 cee eee 339558 Beckman Solution 555 1 qt 0 cece eee eee eee 339555 Rotor cleaning brush 0 0 0 cece eee eee eee 339379 Centering tool for replacing overspeed disk 331325 ULTRACENTRIFUGE ROTOR WARRANTY All Beckman Coulter ultracentrifuge Fixed Angle Vertical Tube Near Vertical Tube Swinging Bucket and Airfuge rotors are warranted against defects in materials or workmanship for the time periods indicated below subject to the Warranty Conditions stated below Preparative Ultracentrifuge Rotors 5 years No Proration Analytical Ultracentrifuge Rotors 5 years No Proration ML and TL Series Ultracentrifuge ROOTS eae op Ps esata idle gad an Pais Ree 5 years No Proration Airfuge Ultracentrifuge Rotors 1 year No Proration For Zonal Continuous Flow Component Test and Rock Core ultracentrifuge rotors see separate warranty Warranty Conditions as applicable 1 This warranty is valid for the time periods indicated above from the date of shipment to the original Buyer by Beckman Coulter or an authorized Beckman Coulter representative 2 This warranty extends only to the original Buyer and may not be assigned or extended to a third person without written consent of Beckman Coulter 3 This
21. by an appropriate Beckman Coulter Service Agreement Contact your local Beckman Coulter office for details of service coverage or cost Before shipping contact the nearest Beckman Coulter Sales and Service office and request a Returned Goods Authorization RGA form and packaging instructions Please include the complete rotor assembly with buckets lid handle tube cavity caps etc A SIGNED STATEMENT THAT THE ROTOR AND ACCESSO RIES ARE NON RADIOACTIVE NON PATHOGENIC NON TOXIC AND OTHERWISE SAFE TO SHIP AND HANDLE IS REQUIRED Beckman Coulter Worldwide Biomedical 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 1 905 819 1234 Fax 1 905 819 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 CZECH REPUBLIC Beckman Coulter Prague Radiova 1 102 27 Prague 10 Czech Republice Telephone 420 267 00 85 13 Fax 420 267 00 83 23 EASTERN EUROPE MIDDLE EAST NORTH AFRICA Beckman Coulter International S A 22 Rue Juste Olivier
22. cturers may fit in the Type 50 2 Ti rotor their safety in this rotor cannot be ascertained by Beckman Coulter Use of other manufacturers components or accesso ries in the Type 50 2 Ti rotor may void the rotor warranty and should be prohibited by your laboratory safety officer Only the components and accessories listed in this publication should be used in this rotor Read the Safety Notice page at the front of this manual before using the rotor 1 Make sure that the rotor and lid are clean and show no signs of corrosion or cracking 2 Make sure that the rotor is equipped with the correct overspeed detection disk If the disk is missing or damaged replace it according to the instructions in Rotors and Tubes 3 Check the chemical compatibilities of all materials used refer to Appendix A in Rotors and Tubes 4 Verify that the tubes and bottles being used are listed in Table 1 For runs at other than room temperature refrigerate or warm the rotor beforehand for fast equilibration 1 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 Type 50 2 Ti Rotor OPERATION 2 Load the filled containers symmetrically into the rotor Refer to TUBES AND BOTTLES on page 7 for information about containers If fewer than twelve tubes are being run they must be arranged
23. damage components no guarantee of decontamination is expressed or implied Consult your laboratory safety officer regarding the proper decontamination methods to use If the rotor or other components are contaminated with toxic or pathogenic materials follow appropriate decontamination procedures as outlined by your laboratory safety officer 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 In U S contact Biodex Medical Systems Shirley New York internationally contact the U S office to find the dealer closest to you 22 Type 50 2 Ti Rotor STERILIZATION AND DISINFECTION e The rotor and all rotor components can be autoclaved at 121 C for up to an hour Remove the lid from the rotor and place the rotor lid and spacers in the autoclave upside down 121 C Ethanol 70 or hydrogen peroxide 6 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 Com While Beckman Coulter has tested these methods and found that they do not damage the rotor or components no guarantee of sterility or disinfection is expressed or implied When sterilization or
24. e 7 Rotor parts subject to wear including but not limited to rotor O rings VTi NVT TLV MLN and TLN rotor tube cavity plugs and gaskets tubing tools optical overspeed disks bear ings seals and lubrication are excluded from this warranty and should be frequently inspected and replaced if they become worn or damaged 8 Keeping a rotor log is not mandatory but may be desirable for maintenance of good laboratory practices Repair and Replacement Policies 1 Ifa Beckman Coulter rotor is determined by Beckman Coulter to be defective Beckman Coulter will repair or replace it subject to the Warranty Conditions A replacement rotor will be warranted for the time remaining on the original rotor s warranty 2 Ifa Beckman Coulter centrifuge is damaged due to a failure of a rotor covered by this warranty Beckman Coulter will supply free of charge i all centrifuge parts required for repair except the drive unit which will be replaced at the then current price less a credit determined by the total number of revolutions or years completed provided that such a unit was manufactured or rebuilt by Beckman Coulter and ii if the centrifuge is cur rently covered by a Beckman Coulter warranty or Full Service Agreement all labor necessary for repair of the centrifuge 3 If a Beckman Coulter rotor covered by this warranty is dam aged due to a malfunction of a Beckman Coulter ultracentrifuge covered by an Ultracentrifuge Syste
25. e rotor lid If disassembly reveals evidence of leakage you should assume that some fluid escaped the rotor Apply appropriate decontamination procedures to the centrifuge and accessories Never exceed the maximum rated speed of the rotor and labware in use Refer to the section on RUN SPEEDS and derate the run speed as appropriate Do not use sharp tools on the rotor that could cause scratches in the rotor surface Corrosion begins in scratches and may open fissures in the rotor with continued use Type 50 2 Ti Rotor TYPE 50 2 Ti ROTOR U S Pat No 3 961 745 U S Pat No 3 901 434 British Pat No 1 465 099 Canadian Pat No 1 001 597 i German Pat No 2 448 199 Axis of Rotation SPECIFICATIONS Maximum speed 0 0 00 cece eee iess 50 000 rpm Density rating at maximum speed 000 1 2 g mL Relative Centrifugal Field at maximum speed At rmax 107 9 mm 0 eee ee 302 000 x g At ray 81 2 mm eiiie ai n eee 227 000 x g At rmin 54 4 mm 0 eee ee 152 000 x g k factor at maximum speed 00 eee eee eee 69 Conditions requiring speed reductions see RUN SPEEDS Number of tube cavities 00 eee ee 12 Available tubes 23 44 45 dte tow cidade bee beaks see Table 1 Nominal tube dimensions largest tube 25 x 89 mm Nominal tube capacity largest tube 00 0 39 mL Nominal rotor capacity 0 0 0
26. e the appropriate removal tool listed in the SUPPLY LIST to remove the spacers and tubes The Type 50 2 Ti rotor uses the tubes and bottles listed in Table 1 Be sure to use only those items listed and to observe the maximum speed limits and fill volumes shown Maximum fill volume is the maximum amount that can be centrifuged in the container listed Refer to Appendix A in Rotors and Tubes for chemical compatibilities of tube bottle and accessory materials Type 50 2 Ti Rotor Table 1 Available Tubes and Bottles for the Type 50 2 Ti Rotor Use only the items listed here and observe fill volumes and maximum run speeds Tube Required Accessory Max Speed Part Max Part RCF Dimensions Description Number Fill Volume Description Number k Factor 25 x 89mm Quick Seal 342414 39 mL red aluminum 342699 50 000 rom polyallomer spacer 302 000 x g 69 25 x 89mm Quick Seal 344326 38 5 mL red aluminum 342699 50 000 rpm Ultra Clear spacer 302 000 x g 69 25x 89mm thinwall 326823 38 5 mL red anodized 331151 50 000 rpm polyallomer cap 302 000 x g 69 25 x 89mm Ultra Clear 344058 38 5 mL red anodized 331151 50 000 rpm cap 302 000 x g 69 25x 89mm stainless steel 301112 38 5 mL aluminum cap 302133 37 000 rpm 165 000 x g 127 25x 79mm Quick Seal 344623 33 mL Delrin spacer 344635 50 000 rom polyallomer 302 000 x g 68 25x 77mm OptiSeal 361625 32 4 mL Ultem4 spacer 361669 50 000
27. filled tube may be centrifuged no faster than 36 500 rpm Gradient curves not shown in Figure 3 can be interpolated A tube full of the 1 615 g mL CsCl solution may be centrifuged no faster than 30 000 rpm TYPICAL EXAMPLES FOR DETERMINING CsCl RUN PARAMETERS At Speed Floating Components Bands Pelleted Material At Rest in Rotor Upright Example A Knowing homogeneous CsCl solution density 1 52 g mL and approximate particle buoyant densities 1 50 and 1 55 g mL where will particles band 1 In Figure 2 find the curve that corresponds to the required run temperature 4 C and fill volume three quarters full The maximum allowable rotor speed is determined from the point where this curve intersects the homogeneous CsCl density 40 000 rpm 2 In Figure 3 sketch in a horizontal line corresponding to each particle s buoyant density 3 Mark the point in the figure where each particle density intersects the curve corresponding to the selected run speed and temperature 4 Particles will band at these locations across the tube diameter at equilibrium during centrifugation In this example particles will band about 86 and 90 mm from the axis of rotation about 4 mm of centerband to centerband separation at the rotor s 24 degree tube angle When the tube is held upright there will be about 4 38 mm of centerband to centerband separation This inter band distance dup can be calculated from the formula d
28. ifferent rotor if the k factor of the previous rotor is known For any two rotors a and b t k L a T 5 fh b For more information on k factors see Use of k Factor for Estimating Run Times from Previously Established Run Conditions publication DS 719 The centrifugal force at a given radius in a rotor is a function of speed Comparisons of forces between different rotors are made by comparing the rotors relative centrifugal fields RCF When rotational speed is adjusted so that identical samples are subjected to the same RCF in two different rotors the samples are subjected to the same force The RCF at a number of rotor speeds is provided in Table 2 13 Type 50 2 Ti Rotor 14 Table 2 Relative Centrifugal Fields for the Type 50 2 Ti Rotor Entries in this table are calculated from the formula RCF 1 12r RPM 1000 and then rounded to three significant digits Relative Centrifugal Field x g Rotor Speed At rmax At rav At rmin k rpm 107 9 mm 81 2 mm 54 5 mm Factor 50 000 302 000 227 000 152 000 69 45 000 245 000 184 000 123 000 85 40 000 193 000 146 000 97 500 108 35 000 148 000 111 000 74 600 141 30 000 109 000 81 800 54 800 192 25 000 75 500 56 800 38 100 276 20 000 48 300 36 400 24 400 431 15 000 27 200 20 500 13 700 770 10 000 12 100 9 090 6 090 1734 Calculated for all Beckman Coulter preparative rotors as a measure of the rotor s relative pelleting efficienc
29. k Seal 343665 27 mL Noryl floating 343448 50 000 rom polyallomer spacer 302 000 x g 55 25 x 38mm Quick Seal 344324 15 mL Noryl floating 343448 50 000 rpm Ultra Clear spacer 302 000 x g 39 25 x 38mm Quick Seal 343664 15 mL Noryl floating 343448 50 000 rom polyallomer spacer 302 000 x g 39 16x 76mm Ultra Clear 344085 13 5 mL Delrin adapter 303307 40 000 rpm 179 000 x g aluminum cap 330860 85 16x 76mm thinwall 326814 13 5 mL Delrin adapter 303307 40 000 rpm polyallomer x 179 000 x g aluminum cap 330860 85 16x76mm thickwall 355640 13 5 mL Delrin adapter 303307 40 000 rpm polyallomer m 179 000 x g aluminum cap 338907 85 16x76mm thickwall 355630 13 5 mL Delrin adapter 303307 40 000 rpm polycarbonate 3 179 000 x g aluminum cap 338907 85 16x76mm stainless steel 301108 13 5 mL Delrin adapter 303392 30 000 rpm i r 101 000 x g aluminum cap 346256 126 13x64mm Ultra Clear 344088 6 5 mL Delrin adapter 303392 43 000 rpm 194 000 x g aluminum cap 346256 61 13x64mm thinwall 326820 6 5 mL Delrin adapter 303392 43 000 rpm polyallomer i 194 000 x g aluminum cap 346256 61 a Refer to the section on RUN SPEEDS before using stainless steel tubes continued b Delrin is a registered trademark of E I Du Pont de Nemours amp Company Includes disposable plastic plugs dUltem is a registered trademark of GE Plastics For lower fill volumes 16 5 mL when centrifuged without cap maximum speed is 45 000 rpm f For lower
30. m Service Agreement Beckman Coulter will repair or replace the rotor free of charge 4 If a Beckman Coulter rotor covered by this warranty is damaged due to a failure of a Beckman Coulter tube bottle tube cap spacer or adapter covered under the Conditions of this Warranty Beckman Coulter will repair or replace the rotor and repair the instrument as per the conditions in policy point 2 above and the replacement policy 5 Damage to a Beckman Coulter rotor or instrument due to the failure or malfunction of a non Beckman Coulter tube bottle tube cap spacer or adapter is not covered under this warranty although Beckman Coulter will assist in seeking compensation under the manufacturer s warranty Disclaimer IT IS EXPRESSLY AGREED THAT THE ABOVE WARRANTY SHALL BE IN LIEU OF ALL WARRANTIES OF FITNESS AND OF THE WARRANTY OF MERCHANTABILITY AND BECKMAN COULTER INC SHALL HAVE NO LIABILITY FOR SPECIAL OR CONSEQUENTIAL DAMAGES OF ANY KIND WHATSOEVER ARISING OUT OF THE MANUFAC TURE USE SALE HANDLING REPAIR MAINTENANCE OR REPLACEMENT OF THE PRODUCT Factory Rotor Inspection Service Beckman Coulter Inc will provide free mechanical and metallurgical inspection in Palo Alto California USA of any Beckman Coulter rotor at the request of the user Shipping charges to Beckman Coulter are the responsibility of the user Rotors will be inspected in the user s laboratory if the centrifuge in which they are used is covered
31. mperature Limits e Plastic tubes and bottles have been centrifuge tested for use at temperatures between 2 and 25 C For centrifugation at other temperatures pretest tubes under anticipated run conditions e Ifplastic containers are frozen before use make sure that they are thawed to at least 2 C prior to centrifugation OptiSeal Tubes OptiSeal tubes come with plastic plugs and can be quickly and easily prepared for use With the tube spacer in place the g force during centrifugation ensures a tight reliable seal that protects your samples e Place the tubes in the rack and fill each tube to the base of the stem leaving no fluid in the stem Overfilling the tube can cause spillage when the plug is inserted or can compromise seal integ rity However too much air can cause excessive tube deformation disrupting gradients and sample bands nC Type 50 2 Ti Rotor e Refer to Using OptiSeal Tubes publication IN 189 included in each box of tubes for detailed information on the use and care of OptiSeal tubes Quick Seal Tubes Quick Seal tubes must be sealed prior to centrifugation These tubes are heat sealed and do not need caps however spacers are required on top of the tubes when they are loaded into the rotor e Fill Quick Seal tubes leaving a small bubble of air at the base of the neck Do not leave a large air space too much air can cause excessive tube deformation e Refer to Rotors and Tubes for det
32. ombination must not exceed the limits in Table 1 3 Further speed limits must be imposed when CsCl or other self forming gradient salts are centrifuged as equation 6 does not predict concentration limits speeds that are required to avoid precipitation of salt crystals Solid CsCl has a density of 4 g mL and if precipitated during centrifugation may cause catastrophic rotor failure and instrument damage Figures 2 and 3 together with the description and examples below show how to reduce run speeds when using CsCl gradients 15 Type 50 2 Ti Rotor 16 SELECTING CsCl GRADIENTS ADJUSTING FILL VOLUMES Buffer Gradient Precipitation during centrifugation would alter density distribution and this would change the position of the sample bands Curves in Figures 2 and 3 are provided up to the maximum rated speed of the rotor but note also that tubes or bottles must never be centrifuged faster than the limits in Table 1 I gt NOTE The curves in Figures 2 and 3 are for solutions of CsCl salt dissolved in distilled water only If other salts are present in significant concentra tions the overall CsCl concentration may need to be reduced Rotor speed is used to control the slope of a CsCl density gradient and must be limited so that CsCl precipitation is avoided Speed and density combinations that intersect on or below the curves in Figure 2 ensure that CsCl will not precipitate during centrifugation in the Type
33. r assembly 0 00 0 337901 Rotor handle sien ren he ae eee EO ee BO 337904 O ring large sce csc secs kta t i sR aad EE E ban R 834301 Orring Small ta thet be sok ie ea ren deg a a eee ees 854519 Overspeed disk 50 000 rpm 0 000 000 0 e eee 330336 Tubes bottles and accessories 0 0 00 e eee ee eee see Table 1 OptiSeal tube rack assembly 0 00000 e eee eee 361646 Quick Seal Cordless Tube Topper kit 60 Hz 358312 Quick Seal Cordless Tube Topper kit 50 Hz Europe 358313 Quick Seal Cordless Tube Topper kit 50 Hz Great Britain 358314 Quick Seal Cordless Tube Topper kit 50 Hz Australia 358315 Quick Seal Cordless Tube Topper kit 50 Hz Canada 367803 Tube Topper rack ieonta a a n ENAN 348124 Tool kit for red and blue aluminum caps 331202 includes TOPque wrench cox esa Rep eee La E E EAE oe A EEEE 858121 Socket adapters revan E gata E ERARE de enced 858122 Socket for 19 mm hex nuts 0 ce cee eee eee 858123 Tube removal tool 2 teen eens 301875 Hex driver for 11 mm cap nuts on stainless steel tubes 841883 Floating spacer removal tool 0 0 00 0c eee eee ee 338765 TMDE CAp VISE ris ied sek Or as ER BRR ae Ae EA Rok AE e 305075 25 Type 50 2 Ti Rotor 26 Removal tool polycarbonate bottles with Noryl caps 335381 Tube removal tool Quick Seal and OptiSeal tu
34. rticle s buoyant density 2 Select the curve at the required temperature 20 C and tube volume full that gives the best particle separation 3 Note the run speed along the selected curve 30 000 rpm 4 From Figure 2 select the maximum homogeneous CsCl density in this case 1 67 g mL that corresponds to the temperature and run speed established above These parameters will provide the particle banding pattern selected in Step 2 In this example particles will band at about 72 and 79 mm from the axis of rotation about 7 mm apart When the tube is held upright there will be about 7 66 mm of center of band to center of band separation CARE AND MAINTENANCE MAINTENANCE gt NOTE Do not use sharp tools on the rotor that could cause scratches in the rotor surface Corrosion begins in scratches and may open fissures in the rotor with continued use Rotor ae F ioci Handle e Periodically at least monthly inspect the rotor especially inside 337904 cavities for rough spots or pitting white powder deposits ina a frequently aluminum oxide or heavy discoloration If any 834301 e IY of these signs are evident do not run the rotor Contact your smal Check 854519 Beckman Coulter representative for information about the Field PA n Rotor Inspection Program and the rotor repair center 20 e Regularly lubricate the metal threads in the rotor with a thin even coat of Spinkote lubricant 306812
35. symmetrically in the rotor see Figure 1 Opposing tubes must be filled to the same level with liquid of the same density 3 Complete loading by placing the correct spacers if required over the tubes I gt NOTE Place filled tubes in at least two opposing cavities Make sure that cavities in use have the proper spacers inserted if applicable before installing the rotor lid 4 Put the lid in place and tighten by hand as firmly as possible o o oto O O O O O O O O O o Oe O00 Figure 1 Typical Examples of Symmetric Loading of Twelve or Fewer Tubes 1 Carefully lower the rotor straight down onto the drive hub 2 Refer to the applicable instrument instruction manual for ultracen trifuge operation Type 50 2 Ti Rotor 3 For additional operating information see the following e RUN TIMES page 12 for using k factors to adjust run durations e RUN SPEEDS page 13 for information about speed limitations e SELECTING CsCl GRADIENTS page 16 for methods to avoid CsCl precipitation during centrifugation REMOVAL AND SAMPLE RECOVERY Quick Seal Tube Removal Tool 361668 TUBES AND BOTTLES A CAUTION If disassembly reveals evidence of leakage you should assume that some fluid escaped the rotor Apply appropriate decontamination procedures to the centrifuge and accessories 1 Remove the rotor from the centrifuge by lifting it straight up and off the drive hub 2 Remove the rotor lid 3 Us
36. y in water at 20 C RCF x g 350 000 300 000 250 000 200 000 150 000 100 000 50 000 0 Relative Centrifugal Fields Type 50 2 Ti Rotor Tmax av fmin 0 10 000 20 000 Speed rpm 30 000 40 000 50 000 Type 50 2 Ti Rotor Do not select rotational speeds that exceed the limits in Table 1 In addition speeds must be reduced under the following circumstances 1 If nonprecipitating solutions more dense than 1 2 g mL are centri fuged in plastic tubes or bottles the maximum allowable run speed must be reduced according to the following equation reduced maximum speed 50 000 rpm es 6 where p is the density of tube contents This speed reduction will protect the rotor from excessive stresses due to the added tube load Note however that the use of this formula may still produce maximum speeds that are higher than the limitations imposed by the use of certain tubes or adapters see Table 1 In such cases use the lower of the two speeds 2 For centrifuging solutions of any density in stainless steel tubes and aluminum cap combinations use the following formula to determine the allowable rotor speed based on the tube load reduced maximum speed 50 000 rpm ingen 7 where X the weight in grams of the capped tube plus the liquid contents to be centrifuged In any case the rotor speed when using this tube cap c
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