Instructions for Using Micro Plus Multiwell Plate Carriers

Transcription

1 Litpak GS6-TB-011EA February 2014 Instructions for Using Micro Plus Multiwell Plate Carriers In Beckman Coulter GH-3.8, GH-3.8A, and JS-4.3 Rotors A B C D E F G H SPECIFICATIONS Maximum speed rpm Relative Centrifugal Field* see Table 2 r max mm Maximum carrier load (including base, pad, plate(s), and sample g Maximum solution density g/ml Carrier material aluminum Base material thermoplastic Pad material thermoplastic elastomer * Relative Centrifugal Field (RCF) is the ratio of the centrifugal acceleration at a specified radius and speed (rω 2 ) to the standard acceleration of gravity (g) according to the following formula: RCF = rω g where r is the radius in millimeters, ω is the angular velocity in radians per second (2 π RPM /60), and g is the standard acceleration of gravity (9807 mm/s 2 ). After substitution: RCF = 1.12 r RPM DESCRIPTION Multiwell plates are used for many applications, including culturing small populations of cells, immunoassays, and serial dilution of small liquid volumes. The Micro Plus multiwell plate carrier (see Figure 1) used with the GH-3.8, GH-3.8A, and JS-4.3 rotors can accommodate one deep-well plate, one, two, or three multiwell plates (not to exceed 41.7 mm/1.64 in. depth), or one rack of mini-tubes. Multiwell plate carriers are installed on the rotor yoke pivot pins in place of the A B C D E F G H Deep-well Plate Carrier Figure 1. The Micro Plus Multiwell Plate Carrier, Base, Pad, and Deep-well Plate Pad Base Beckman Coulter, Inc.

2 buckets normally used with the rotor. The carriers are made of aluminium, and are black-anodized for corrosion resistance. A blue plastic base is used to house the multiwell plates. A flexible black plastic pad is used between the base and the plate(s) to provide support during centrifugation. The aluminum carriers are warranted for 7 years; the plastic base and pad are warranted for 1 year. LOADING If only two carriers are run, they must be installed opposite each other in the rotor, and either two other carriers or two buckets must also be run (they need not be loaded) to prevent rotor imbalance. The rotor should never be run with only two carriers or two buckets installed. Refer to Table 1 for labware used with Micro Plus carriers. To ensure optimal performance and stability, the rotor must be loaded symmetrically. For best results you should load opposing carriers with the same type of labware containing the same amounts of fluid of equal density. Do not exceed the rated maximum load for carriers (500 grams, including base, pad, and sample). It is not necessary to completely fill all of the multiwell plate wells; however, partially filled multiwell plates must be balanced with respect to the pivotal axis of the carrier. In multiwell plates, samples should be loaded into the wells symmetrically with respect to the pivotal axis of the carrier (the pivotal axis runs parallel to the crossbar), and opposing carriers should contain similar loads (see Figure 2). CORRECT INCORRECT Examples of Symmetrically Loaded Trays (load opposite trays the same way) Examples of Nonsymmetrically Loaded Trays Pivotal Axis Carrier Plate A B C D E F G H Figure 2. Examples of Correctly and Incorrectly Loaded Carriers. Contents of opposing carriers must be the same and each carrier must be balanced on its pivotal axis. 2

3 Table 1. Labware for Use with Micro Plus Carriers Description Volume Part Number Description Required Accessory Part Number Rack Multiwell plate, 96-well, nonsterile 300 μl/well * (pkg/100) Deep-well polystyrene plate, 96-well, nonsterile 1 ml/well (pkg/24) Cap strip, nonsterile* Cap strip, sterile* Aluminum foil lid* (pkg/10) (pkg/10) (pkg/100) Deep-well polystyrene plate, 96-well, sterile 1 ml/well (pkg/24) Cap strip, nonsterile* Cap strip, sterile* Aluminum foil lid* (pkg/10) (pkg/10) (pkg/100) Deep-well polypropylene plate, 96-well, nonsterile 1 ml/well (pkg/24) Cap strip, nonsterile* Cap strip, sterile* Aluminum foil lid (pkg/10) (pkg/10) (pkg/100) Deep-well polypropylene plate, 96-well, sterile 1 ml/well (pkg/24) Cap strip, nonsterile* Cap strip, sterile* Aluminum foil lid (pkg/10) (pkg/10) (pkg/100) Square-well polypropylene plate 2 ml/well Aluminum foil lid (pkg/100) Microfuge tube, polypropylene, clear 1.5 ml Microfuge tube, polypropylene, clear 1.5 ml Microfuge tube, polypropylene, blue 1.5 ml Microfuge tube, polypropylene, green 1.5 ml Microfuge tube, polypropylene, yellow 1.5 ml Microfuge tube, polypropylene, orange 1.5 ml * Caps are optional. Requires 4-in. soft rubber roller (538618) for installation. 3

4 During a run, carriers swing 90 degrees from their at-rest position. The pivotal axis of a carrier can be imagined as a line extending across the carrier from one pivot pin to the other. If a 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 a run. As a result, extra stress will be placed on the carrier and/ or multiwell plates during the run, increasing the possibility of plate breakage or rotor imbalance. NOTE If symmetrical pelleting is required, use round or V-bottom plates (commercially available from many suppliers) for best results. Load each carrier as follows: 1. To prevent plate breakage during centrifugation, place the flexible plastic pad, ridged side up, into the flat, indented surface of the blue base. 2. Place one deep-well plate or one, two, or three multiwell plates on top of the pad, being careful not to spill the contents. (Remember to load opposing carriers with the same plate configuration.) 3. If you are running more than one 96-well plate, place a 96-cap strip beneath the bottom plate and between the plates. 4. Slide the base, pad, and multiwell plate assembly into the carrier until the base clicks into place. RUN PROCEDURE Refer to the applicable rotor manual and centrifuge instruction manual for operating procedures. NOTE Do not use high brake when multiwell plate carriers are run, as the forces encountered during deceleration may disturb the contents of the plate. Set the centrifuge brake control to either LOW or OFF to coast to a stop. The centrifugal forces at a given radius in the rotor are a function of run speed. Use Table 2 to select a run speed that will give the required RCF. 4

5 Table 2. Relative Centrifugal Fields for the Multiwell Plate Carriers Rotor Speed (rpm) Relative Centrifugal Field ( g) of Samples in Plate (at r max 163 mm) Multiwell plates will break if g-forces are too high. Rotor speed must not exceed 3250 rpm when plate carriers are run. NOTE To prevent overspeeding the carriers, remove the carriers from the rotor immediately after a run is complete. If two buckets and two carriers are used, set the run speed to 3250 rpm (the speed allowed for plate carriers). If the weight of the load in a carrier exceeds 500 grams including base, pad, plates, and sample, use the following square-root reduction formula to determine the allowable rotor speed: speed = (3250 rpm) 500 grams heaviest load in grams Polystyrene plates are subject to cracking at high speeds. Test polystyrene plates using water in place of sample to determine required speed deration. 5

6 CARE AND MAINTENANCE MAINTENANCE Periodically inspect the carriers for rough spots or pitting, white powder deposits (frequently aluminum oxide), or heavy discoloration. If any of these signs are evident, do not run the carrier. Contact your Beckman Coulter representative for information about the Field Rotor Inspection Program and the rotor repair center. The carriers are made of anodized aluminum. Do not use sharp tools on them, as scratches in the anodized surfaces may lead to corrosion, stress cracks, or other damage. Regularly check the condition of the base and rubber pad and do not use them if there is visible damage. CLEANING If spillage has occurred, or if salt solutions or other corrosive materials are used, clean the carrier, base, and pad immediately. Do not allow corrosive materials to dry on rotor components. Under normal conditions, wash the components frequently following the steps below. 1. Use a mild detergent such as Beckman Solution 555 (339555), diluted 10 to 1 with water, and a soft brush to wash the carriers, base, and pad. The Rotor Cleaning Kit (339558) contains two quarts of Solution 555 and brushes that will not scratch the components. NOTE Do not wash the carriers, bases, or pads in a dishwasher. Do not soak these components in detergent solution for long periods, such as overnight. 2. Thoroughly rinse the components with water. 3. Air-dry the components, placing the carrier upside down. Do not use acetone to dry the components. DECONTAMINATION If the carriers become contaminated with radioactive material, they should be decontaminated using a solution that will not damage their anodized surfaces. Beckman Coulter has 6

7 tested a number of solutions and found two that do not harm anodized aluminum: RadCon Surface Spray or IsoClean Solution (for soaking), 1 and Radiacwash. 2 If carriers or other components are contaminated with toxic or pathogenic materials, follow appropriate decontamination procedures as outlined by your laboratory safety officer. Refer to Chemical Resistances (publication IN-175) to evaluate a component s ability to withstand a chemical solution.! CAUTION Strongly alkaline solutions will damage the carrier. While Beckman Coulter has tested these methods and found that they do not damage components, no guarantee of sterility or disinfection is expressed or implied. When sterilization or disinfection is a concern, consult your laboratory safety officer. STERILIZATION AND DISINFECTION The carriers (aluminum parts only) can be autoclaved at 121 C for up to an hour. DO NOT AUTOCLAVE THE PLASTIC BASES OR PADS. Ethanol 3 (70%) may be used on all rotor components, including those made of plastic. Bleach (sodium hypochlorite, 10%) may be used on the plastic bases and pads. Refer to Chemical Resistances for the chemical resistances of tubes and accessories. While Beckman Coulter has tested these methods and found that they do not damage components, no guarantee of sterility or disinfection is expressed or implied. When sterilization or disinfection is a concern, consult your laboratory safety officer. 1 In U.S.A., 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.A., contact Biodex Medical Systems (Shirley, NY); internationally, contact the U.S. office to find the dealer nearest you. 3 Flammability hazard. Do not use in or near operating centrifuges. 7

8 RETURNING A COMPONENT Before returning a carrier component for any reason, prior permission (a Returned Goods Authorization form) must be obtained from Beckman Coulter, Inc. This RGA form, which may be obtained from your local Beckman Coulter sales office, 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 and/or radioactivity. Sterilization 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 outside 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. Customers located outside the United States should contact their local Beckman Coulter office. 8

9 SUPPLY LIST NOTE Publications referenced in this manual can be obtained by calling Beckman Coulter at in the United States, or by contacting your local Beckman Coulter office. Contact Beckman Coulter Sales ( in the United States) or your local Beckman Coulter sales office for detailed information on ordering parts and supplies. For your convenience, a partial list is given below. REPLACEMENT PARTS Micro Plus Carriers (set of 2) Replacement bases for Micro Plus carriers, including base and rubber pad (set of 2 each) Replacement rubber pads (set of 4) SUPPLIES Multiwell plates, tubes, and accessories see Table 1 Rotor Cleaning Kit Solution 555 (1 qt) TRADEMARKS Beckman Coulter, the stylized logo, and Microfuge are trademarks of Beckman Coulter, Inc. and are registered with the USPTO. All other trademarks, service marks, products, or services are trademarks or registered trademarks of their respective holders. 9

10 Beckman Coulter, Inc. 250 S. Kraemer Blvd. Brea, California U.S.A. Sales and Service: Internet: Printed in USA 2014 Beckman Coulter, Inc. All rights reserved