Thermo Fisher Scientific AH-629 Swinging Bucket Rotor User manual

54276-9 March 2010

Thermo Fisher Scientific

AH-629

Instruction Manual

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free and assumes no responsibility and will not be liable for any errors, omissions, damage or loss that might

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Sale shall govern all conflicting information between the two documents.

Release history: 54276-9 printed in March 2010.

For Research Use Only. Not for use in diagnostic procedures.

Thermo Scientific AH-629 i

T

Thermo Scientific AH-629 Aluminum Swinging Bucket Ultracentrifuge Rotor ..............i

Important Safety Information .................................................................................................. ii

Chapter 1 DESCRIPTION ........................................................................................................................ 1-1

Rotor Description .................................................................................................... 1-2

Rotor Specifications ................................................................................................. 1-2

Accessories ............................................................................................................... 1-3

Chapter 2 SPECIAL OPERATING CONSIDERATIONS ........................................................................ 2-1

Derating Rotor Speed .............................................................................................. 2-2

Compartment Loads in Excess of Design Mass ........................................................ 2-2

Precautions to Prevent Precipitation of Cesium Chloride ......................................... 2-2

Critical Speed .......................................................................................................... 2-4

Chapter 3 OPERATION ............................................................................................................................. 3-1

Prerun Safety Checks ............................................................................................... 3-2

Rotor Precool .......................................................................................................... 3-2

Chemical Compatibility .......................................................................................... 3-2

Relative Centrifugal Force (RCF) Determination .................................................... 3-3

Calculation of Sedimentation Times in Aqueous (Non-Gradient) Solutions ............ 3-4

AH-629 RCF Values and K Factors ......................................................................... 3-4

Tube Filling and Bucket Loading Procedure .......................................................... 3-10

Rotor Balancing ..................................................................................................... 3-11

Bucket Attachment ................................................................................................ 3-12

Rotor Installation ................................................................................................... 3-12

Centrifuge/Rotor Log ............................................................................................ 3-13

Chapter 4 MAINTENANCE ..................................................................................................................... 4-1

Corrosion ................................................................................................................ 4-2

Cleaning and Decontamination ............................................................................... 4-2

Inspection ................................................................................................................ 4-3

Overspeed Decal Replacement ................................................................................. 4-3

Storage ..................................................................................................................... 4-3

Service Decontamination Policy .............................................................................. 4-4

Chemical Compatibility Chart ...............................................................................A-1

Contact Information ................................................................................................B-1

Table of contents

Thermo Scientific AH-629 i

P

This manual is a guide for the use of

Thermo Scientific AH-629 Aluminum Swinging

Bucket Ultracentrifuge Rotor

Data herein has been verified and is believed adequate for the intended use of the centrifuge. Because

failure to follow the recommendations set forth in this manual could produce personal injury or

property damage, always follow the recommendations set forth herein. Thermo Fisher Scientific does

not guarantee results and assumes no obligation for the performance of rotors or other products that

are not used in accordance with the instructions provided. This publication is not a license to operate

under, nor a recommendation to infringe upon, any process patents.

Publications prior to the Issue Date of this manual may contain data in apparent conflict with that

provided herein. Please consider all data in this manual to be the most current.

NOTES, CAUTIONS, and WARNINGS within the text of this manual are used to emphasize

important and critical instructions.

WARNING informs the operator of a hazard or unsafe practice that could result in personal injury,

affect the operator's health, or contaminate the environment.

CAUTION informs the operator of an unsafe practice that could result in damage of equipment.

NOTE highlights essential information.

CAUTION and WARNING are accompanied by a hazard symbol and appear near the

information they correspond to.

Preface

ii AH-629 Thermo Scientific

Important Safety Information

Certain potentially dangerous conditions are inherent to the use of all centrifuge rotors. To ensure safe

operation of this rotor, anyone using it should be aware of all safe practices and take all precautions

described below and throughout this manual.

WARNING

When using radioactive, toxic, or pathogenic materials, be aware of all characteristics of

the materials and the hazards associated with them in the event leakage occurs during

centrifugation. In the event of a rotor failure, neither the centrifuge nor the rotor can

protect you from particles dispersed in the air. To protect yourself, we recommend

additional precautions be taken to prevent exposure to these materials, for example, use of

controlled ventilation or isolation areas.

Always be aware of the possibility of contamination when using radioactive, toxic, or

pathogenic materials. Take all necessary precautions and use appropriate decontamination

procedures if exposure occurs.

Never use any materials capable of producing flammable or explosive vapors, or creating

extreme exothermic reactions.

Never exceed the maximum rated speed of the installed rotor; to do so can cause rotor

failure.

Always reduce (derate) rotor speed as instructed in this manual whenever:

•the rotor speed/temperature combination exceeds the solubility of the gradient

material and causes it to precipitate.

•the compartment load exceeds the maximum allowable compartment load specified.

See Chapter 3, Operation. Failure to reduce rotor speed under these conditions can

cause rotor failure.

Centrifuges routinely deal with high energy levels and could move suddenly in the

unlikely event of rotor failure. During centrifuge operation, never lean on or move the

centrifuge, keep the surrounding area clear of objects (including all hazardous materials),

and do not work on top of or next to the centrifuge.

Do not attempt to open the chamber door when the rotor is spinning; never override or

otherwise disable any of the safety systems of the centrifuge.

Preface

Thermo Scientific AH-629 iii

CAUTION

Do not expose aluminum rotor components to: strong acids, bases, alkaline laboratory

detergents, liquid chlorine bleach or salts (chlorides) of heavy metals such as cesium, lead,

silver, or mercury. Use of these materials with aluminum can cause a chemical reaction that

initiates corrosion.

Do not operate or precool a rotor at the critical speed, as this will have a detrimental effect

on centrifuge component life. See Chapter 3, Operation.

Do not operate the rotor unless it is symmetrically balanced as described in this manual.

Operating the rotor out of balance can cause damage to the rotor chamber. See Chapter 3,

Operation.

Always maintain the centrifuge in the recommended manner. All accessories must be clean

and inspected prior to each run: do not use rotor showing signs of corrosion or cracking.

See Chapter 4, Maintenance.

Do not autoclave the aluminum rotor body or the aluminum cap assemblies to

temperatures in excess of 100°C.

Thermo Scientific AH-629 1-1

1

DESCRIPTION

This manual provides you with the information you will need to operate and maintain your Thermo

Scientific A-629 Swinging Bucket Ultracentrifuge Rotor. If you encounter any problem concerning

either operation or maintenance that is not covered in the manual, please contact our Marketing

Technical Group for assistance. In the United States, call toll free 1-866-9THERMO. Outside the

United States, contact the distributor or agent for Thermo Fisher Scientific products. Thermo Fisher

Scientific product information is available on our internet web site at

http://www.thermo.com/centrifuge .

Contents

•“Rotor Description” on page 1-2

•“Rotor Specifications” on page 1-2

•“Accessories” on page 1-3

1 DESCRIPTION

Rotor Description

1-2 AH-629 Thermo Scientific

Rotor Description

The AH-629 is an aluminum swinging bucket ultracentrifuge rotor that can produce up to six sets of

data during a single run at speeds up to 29,000 rpm.1 Each titanium rotor bucket will hold a

centrifuge tube having a nominal fluid capacity of either 36 ml, 20 ml, or 17 ml depending on the

buckets purchased. The buckets are sealed during operation by an O-ring and an aluminum bucket

cap. The buckets attach to the rotor body, each at its own numbered position. During a centrifuge

run, they swing out horizontally and return to a vertical position during deceleration. A disc with

alternating black and reflective segments attached to the bottom of the rotor provides overspeed

protection.

Rotor Specifications

General

The following specifications apply to the rotor when used with 36 ml, 20 ml, or 17 ml buckets.

Rotor with 36 ml Buckets:

1Speed in revolutions per minute (rpm) is related to angular velocity, w, according to the following:

Where w = rad/s. All further references in this manual to speed will be designated as rpm.

ω(rpm)=2π

60

------

⎝⎠

⎛⎞ rpm()0.10472()=

Table 1-1.Rotor Specifications

Rotor Type Swinging Bucket

Maximum Speed 29,000 rpm*

*With tubes filled with a homogeneous solution having an average density of 1.2 g/ml or less

Number of Buckets 6

Rotor Diameter 21 cm (8.26 in)

Critical Speed 1100 rpm

Table 1-2.Rotor Specifications with 36 ml Bucket

Relative Centrifugal Force (RCF) at maximum speed:

at rminimum (7.21 cm)

at raverage (11.655 cm)

at rmaximum (16.10 cm)

67,731

109,487

151,243

K Factor at maximum speed 242

Total volume capacity (nominal) 216 ml

Tube compartment diameter 2.5 cm (1.0 in)

Tube length (nominal) 8.9 cm (3.5 in)

Maximum compartment mass (includes bucket weight) 156.6 g

Rotor weight including buckets 6.4 kg (14.1 lb)

1 DESCRIPTION

Accessories

Thermo Scientific AH-629 1-3

Rotor with 20 ml Buckets:

Rotor with 17 ml Buckets:

Accessories

The accessories supplied with the AH-629 Rotor ordered with complete accessories are listed in Table

1-5. Catalog numbers are: 54283 (with 17 ml buckets), 54311 (with 20 ml buckets), or 54282 (with

36 ml buckets). The AH-629 Rotor can also be purchased with basic accessories only, which includes

all items listed in Table 1-5 except the tubes. The catalog numbers for the rotor with basic accessories

only are: 54285 (17 ml buckets), 54308 (20 ml buckets), and 54284 (36 ml buckets).

Table 1-3. Rotor Specifications with 20 ml Bucket

Relative Centrifugal Force (RCF) at maximum speed:

at rminimum (7.21 cm) 67,731

at raverage (10.07 cm) 94,597

at rmaximum (12.93 cm) 121,464

K Factor at maximum speed 176

Total volume capacity (nominal) 120 ml

Tube compartment diameter 2.5 cm (1.0 in)

Tube length (nominal) 5.1 cm (2.0 in)

Maximum compartment mass (includes bucket weight) 115.0 g

Rotor weight including buckets 6.1 kg (13.5 lb)

Table 1-4. Rotor Specifications with 17ml Bucket

Relative Centrifugal Force (RCF) at maximum speed:

at rminimum (6.45 cm) 60,591

at raverage (11.52 cm) 108,219

at rmaximum (16.59 cm) 155,846

K Factor at maximum speed 284

Total volume capacity (nominal) 102 ml

Tube compartment diameter 1.6 cm (0.63 in)

Tube length (nominal) 10.2 cm (4.0 in)

Maximum compartment mass (includes bucket weight) 130.1 g

Rotor weight including buckets 6.2 kg (13.7 lb)

Table 1-5. Accessories Supplied

Quantity Catalog Number Description

1 set 52030 Buckets, 17 ml; or

54293 Buckets, 20 ml; or

52035 Buckets, 36 ml

4 boxes of 50 03126*Tubes, Polyallomer (17 ml buckets); or

1 DESCRIPTION

Accessories

1-4 AH-629 Thermo Scientific

8 boxes of 25 03288*Tubes, Polyallomer (20 ml buckets); or

8 boxes of 25 03141*Tubes, Polyallomer (36 ml buckets)

6 63712*O-rings (17 ml buckets) or

64667*O-rings (20 ml and 36 ml buckets)

1 51362 Overspeed Decal, 29 000 rpm (extra)

1 52240 Bucket Rack

1 51942 Rotor Storage Stand

1 65937 Vacuum Grease

1 61556 Lubricant

1 52384 Ultraspeed Centrifuge/Rotor Log Book

1 54276 AH-629 Rotor Instruction Manual

*The catalog number supplied depends on the size of the buckets ordered with the rotor. No tubes are supplied with catalog numbers

54285, 54308, and 54284.

Table 1-5.Accessories Supplied

Quantity Catalog Number Description

Thermo Scientific AH-629 2-1

2

SPECIAL OPERATING CONSIDERATIONS

This chapter provides special operating considerations for Basic Operation.

Contents

•“Derating Rotor Speed” on page 2-2

•“Compartment Loads in Excess of Design Mass” on page 2-2

•“Precautions to Prevent Precipitation of Cesium Chloride” on page 2-2

•“Critical Speed” on page 2-4

2 SPECIAL OPERATING CONSIDERATIONS

Derating Rotor Speed

2-2 AH-629 Thermo Scientific

Derating Rotor Speed

Because of the stresses that the rotor body and buckets must withstand during centrifugation, it is

necessary to eventually derate the maximum operating speed of the rotor. Specifically, the maximum

speed of 29,000 rpm must be derated to 26,000 rpm after the rotor has been used for 1000 runs. To

know when the maximum rotor speed must be lowered, all runs should be recorded in the Ultraspeed

Centrifuge/Rotor Log Book (supplied with the rotor)..

Compartment Loads in Excess of Design Mass

The maximum run speed (29,000 rpm) is based on the recommended design mass that has been

established for the rotor, representing the maximum mass that can be carried in each rotor bucket at

topspeed operation. Total contents of each bucket, including the bucket, specimen and tube, should

not exceed the recommended figure unless the rotor speed is reduced proportionately.

The design mass (that is, the weight of the bucket, tube and contents) for the AH-629 rotor at

29,000 rpm is: 156.6 g (36 ml buckets), 115.0 g (20 ml buckets), or 130.1 g (17 ml buckets). These

figures are based on the use of a Thermo Fisher Scientific thinwall polyallomer tube filled with a

liquid at 1.2 specific gravit.y If the specific gravity of the solution is greater than 1.2 g, use the

following formula to determine the reduced speed:

* 26 000 rpm if rotor has been derated.

This formula does not apply when using gradients that can precipitate.

Precautions to Prevent Precipitation of Cesium Chloride

Reducing Speed to Prevent Precipitation

Maximum speed must be reduced for an average fluid density greater than 1.2 g/ml (square-root

reduction) to prevent excessive hydraulic pressure in the bucket. Although the standard formula (see

previous paragraph) pertains to sucrose and similar gradient materials, it will not prevent

precipitation of crystals when materials such as cesium chloride (CsCl) are used in an ultracentrifuge.

Note After the maximum speed has been derated, the new, lowered maximum speed should be used

in all calculations and during all operations for the ensuing 1000 runs. (These runs should also be

recorded in the Ultraspeed Centrifuge/Rotor Log Book.) In addition, the overspeed decal on the

bottom of the rotor must be replaced with a new decal (Catalog No. 51365) for the lowered speed.

WARNING After the rotor has been used for 1000 runs at the derated speed, the rotor

body and any set of buckets used with the rotor body should no longer be used.

Reduced Speed 29000 1.2 g/ml

Average Fluid Density

---------------------------------------------------=

WARNING Rotor speed must be reduced according to the following instructions for

applications where precipitation is possible. Failure to do so can cause rotor failure with

subsequent sample loss and damage to your centrifuge.

2 SPECIAL OPERATING CONSIDERATIONS

Precautions to Prevent Precipitation of Cesium Chloride

Thermo Scientific AH-629 2-3

When solid crystalline CsCl forms, it places a density of 4 g/ml at the bottom of each bucket. This

density is dangerously high and can cause the rotor to fail, with subsequent sample loss. Therefore,

CsCl solutions must be run at reduced speeds to avoid this precipitation. The allowable speed is

determined by the average density of the CsCl solution and the run temperature. Saturation limits of

CsCl in aqueous solutions are temperature dependent. The solubility limit of 1.86 g/ml at 25°C

becomes 1.81 g/ml at 5°C. CsCl precipitation curves are given for each size bucket for specific average

densities at 5°C and 25°C that will prevent both precipitation and hydraulic pressure. These curves

should be used to determine the maximum operating speed of the rotor. Curves are also given that

show the standard speed (square root) reductions to avoid only excessive hydraulic pressure at 5°C and

at 25°C. For example, standard speed reduction would allow you to run a full tube of solution having a

homogenous density of 1.65 g/ml at 24,700 rpm in any bucket at both the 5°C and 25°C

temperatures. However, you can see that a CsCl solution will precipitate at this speed in the 17 ml and

36 ml bucket at both 5°C and 25°C and in the 20 ml buckets at 5°C.

The graphs show that the highest speed you can run a full tube of a CsCl solution having a density of

1.65 g/ml is:

Similarly, if you are to run the same CsCl solution (1.65 g/ml) in a 3/4-filled tube, the highest speed

you can run this solution at is:

The Gradient Shape

The curves in Figures 2-2, 2-6, 2-8, 2-10 and 2-12 show the gradient shape at equilibrium for tubes

filled with a CsCl solution using the homogenous density and speeds selected from the CsCl

precipitation curves. The shape of a gradient produced in a partially filled tube can be determined by

using lines for 3/4, 1/2 and 1/4 fills. For example, Figure 2-1 indicates that a tube 3/4-filled with a 1.65

g/ml homogenous CsCl solution using 17 ml buckets cannot be run any faster than 16,000 rpm at

5°C. Figure 2-2 shows the shape of the gradient curve which can be interpolated between the 15,000

rpm curve and 17,500 rpm curve. The gradient profile of this tube, 1.53 g/ ml at the meniscus to 1.81

g/ml at the bottom of the tube. To locate a band of particles having a specific density of 1.58 g/ml in a

3/4-filled tube using 17 ml buckets run at 16,000 rpm at 5°C, the particles will band approximately 3

ml below the 3/4 fill line of the tube (Figure 2-2). Similarly, Figure 2-7 shows that the same solution at

25°C using the 17 ml buckets cannot be run any faster than 21,000 rpm. Figure 2-8 shows the shape of

the gradient curve which can be interpolated between the 20,000 rpm curve and 22,500 rpm curve.

14,400 rpm at 5°C using 17 ml buckets (Figure 2-1)

22,300 rpm at 5°C using 20 ml buckets (Figure 2-3)

15,600 rpm at 5°C using 36 ml buckets (Figure 2-5)

18,700 rpm at 25°C using 17 ml buckets (Figure 2-7)

24,700 rpm at 25°C using 20 ml buckets (Figure 2-9)

20,300 rpm at 25°C using 36 ml buckets (Figure 2-11)

16,000 rpm at 5°C using 17 ml buckets (Figure 2-1)

25,400 rpm at 5°C using 20 ml buckets (Figure 2-3)

17,500 rpm at 5°C using 36 ml buckets (Figure 2-5)

21,000 rpm at 25°C using 17 ml buckets (Figure 2-7)

26,000 rpm at 25°C using 20 ml buckets (Figure 2-9)

22,600 rpm at 25°C using 36 ml buckets (Figure 2-11)

2 SPECIAL OPERATING CONSIDERATIONS

Critical Speed

2-4 AH-629 Thermo Scientific

The gradient profile of this tube, 1.48 g/ml at the meniscus to 1.86 g/ml at the bottom of the tube.

To locate a band of particles having a specific density of 1.58 g/ml in a 3/4-filled tube using 17 ml

buckets run at 21,000 rpm at 25°C, the particles will band approximately 4 ml below the 3/4 fill line

of the tube (Figure 2-8). Similarly, the example illustrated in Figures 2-4, 2-6, 2-10 and 2-12 shows

the gradient profile using the same homogenous CsCl solution (1.65 g/ml) in a 3/4-filled tube run at

the 5°C and 25°C temperatures for 20 ml buckets and 36 ml buckets.

Critical Speed

The critical speed is that speed at which any rotor imbalance will produce a driving frequency equal

to the resonant frequency of the rotating system (that is, rotor and the centrifuge drive). At this

speed, the rotor may produce large amplitude vibrations which can be felt in the instrument frame.

Mass imbalance will contribute to increased vibration intensity at the critical speed. Avoid operating

the rotor at the critical speed range which is 1100 rpm for the AH-629 rotor.

Note The gradient shape curves can be used for any fill volume up to the maximum volume of the

tube, unless noted otherwise, in which case, examples are given for partial fills. In these cases,

gradient shapes for other fill volumes at those speeds must be derived by interpolation..

CAUTION Continued operation at the critical speed will have a detrimental effect on

centrifuge component life.

2 SPECIAL OPERATING CONSIDERATIONS

Critical Speed

Thermo Scientific AH-629 2-5

Figure 2-1. CsCl Precipitation Curve, 17 ml Buckets, 5°C

2 SPECIAL OPERATING CONSIDERATIONS

Critical Speed

2-6 AH-629 Thermo Scientific

Figure 2-2. CsCl Gradients at Equilibrium, 17 ml Buckets, 5°C

2 SPECIAL OPERATING CONSIDERATIONS

Critical Speed

Thermo Scientific AH-629 2-7

Figure 2-3. CsCl Precipitation Curve, 20 ml Buckets, 5°C

2 SPECIAL OPERATING CONSIDERATIONS

Critical Speed

2-8 AH-629 Thermo Scientific

Figure 2-4. CsCl Gradients at Equilibrium, 20 ml Buckets, 5°C

2 SPECIAL OPERATING CONSIDERATIONS

Critical Speed

Thermo Scientific AH-629 2-9

Figure 2-5. CsCl Precipitation Curve, 36 ml Buckets, 5°C

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Thermo Fisher Scientific AH-629 Swinging Bucket Rotor User manual

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