Basic Centrifugation

Training
Tubes & Bottles
How do we separate Particles?
• There are many impressive
sounding terms and procedures, but
trust me, it’s simple. All matter has
certain properties that help us to
describe it. Lets visualize an object.
Describe the objects properties to
me in general terms.
Isopycnic & Differential
Centrifugation Methods
 Centrifugal Calculations
• G Force or Relative Centrifugal Field-RCF
2
RCF=1.12 r(RPM/1000)

• k factors are an estimate of the efficiency of a

rotor and can be used to estimate run times
in different rotors

− 5
2 . 53 x10 x ln( r min )
k= r max
( RPM )2
1000
Centrifuge Applications
• Subcellular Fractions
• Nucleic Acids--DNA, RNA
• Cells
• Bacteria
• Virus/Phage/Vectors
• Proteins
• Phase separations
• Binding studies
• Membranes
• Lipoproteins
• Filtering
 Top 10 Rotors
(and the tubes used in them)
• Numbers taken from 1994 Sales Records
Ultra’s High Speeds
1. SW 41 JA-20
2. SW 28 JA-14
3. Ty 70.1 Ti JA-10
4. Ty 70 Ti JA-17
5. Ty 45 Ti JS-13.1
6. SW 55 Ti JLA10.500
7. NVT 90 JA-21
8. NVT 65 JA-20.1
9. SW 40 Ti JA-18.1
10. NVT 65.2 JA-12
 Tube & Bottle Materials
• Clear Materials
Ø Ultra-Clear--PETG, Polycarbonate, Cellulose Propianate
Ø Glass tubes--Corex, Pyrex, Kimax

• Translucent Materials
Ø Polyallomer--Copolymer of propylene & ethylene, Eastman trade name so
similar materials often are sold as Polypropylene
Ø Polypropylene
Ø Polyethylene
Ø Teflon--PTFE

• Tinted Materials
Ø Polysulfone
Ø Polycarbonate has a slight tint
Ø PES, PEI used only in aerosolve canisters, and various covers
 The Ideal Tube Material
• Optically Clear
• Complete chemical resistance to even the harshest
solvents (Phenol, Chloroform, toluene, etc.)

• Ridgid enough to withstand any level of centrifugal

force at any fill level
and any temperature!

• Easily sliced, punctured, wettable

if need be
 Polyallomer
Polyallomer is a trademark of Eastman Kodak and is a random block
copolymer comprised of 97% polypropylene and 3% polyethylene. Similar
products from other producers are called simply polypropylene. As a
polyolefin it is waxlike and translucent, strong and flexible with a high
crystalline structure. This high crystallinity of polyolefins provide good
chemical resistance to density gradient materials and is satisfactory for use
with most acids, bases, alcohols and solvents. It can be autoclaved but
does not perform well with organic solvents. There is very little charge on
the surface of the tube so it does not typically bind biological material.
Recent development of material has yielded a polymer which is almost
transparent when in contact with fluid. This clearer material is used in
OptiSeal tubes!

• Advantages u Disadvantages
1. Good Strength
2. Good chemical resistance 1. Translucent
3. Flexible, easily punctured/sliced 2. Does not tolerate organic
4. Very inert solvents
5. Autoclavable
 Polypropylene
Very similar to polyallomer, it is a member of the polyolefin family
so again it is waxy and translucent. Recent innovations have
resulted in good contact clarity (tube is virtually clear with fluid in
it) as found in the OptiSeal tubes. Good for temperature range
from 2 degrees C to 40 degrees. It has chemical properties
essentially equal to Polyallomer. Can be used with solutions of pH
11 or less.

• Advantage u Disadvantage
1. Versatile
1. Translucent
2. Dimensionally stable over
wide temperature range 2. Does not resist
3. Inert organic solvents
4. Autoclavable
 Ultra-Clear --PETG
Polyesterptherathalate glycol modified. Ulta-Clear is Transparent to aid in
visualizing specific locations of bands or pellets in the tube. It is made of a
tough and somewhat flexible thermoplastic. It has poor chemical resistance
because of it’s amorphous structure (this characteristic makes all clear tubes
have limited chemical resistance). Should not be used with solutions of pH
greater than 8 and can’t be autoclaved. Chemical (often called cold
sterilization) is used such as immersion in 10% hydrogen peroxide for 30
minutes or irradiation. Usable temperature range is 2 to 25 degrees C. Not
recommended for freezing or boiling samples.

• Advantages u Disadvantages

1. Optically Clear 1. Poor Chemical

2. Strong, Ridgid resistance
3. Inert 2. Not autoclavable
3. Slightly difficult to
puncture or slice
 Polycarbonate
Polycarbonate is an extremely ridgid, clear, almost glass like
material. Being clear it is amorphous in structure and has poor
chemical resistance to alkaline solutions, alcohols, solvents, and
any solution of pH 9.0 or higher. Chemical sterilization is
recommended as autoclaving reduces the useful life of the tubes
and bottles. Polycarbonate is often reused and is susceptible to
scratches from removal of pellets from the bottom of the tube. The
scratches weaken the plastic by acting as a stress concentrator.

• Advantages u Disadvantages

1. Strong, clear & durable 1. Weakened by scratches

2. Inert 2. Poor chemical resistance
3. Reusable 3. Weakened by
autoclaving
 Cellulose Propionate
Cellulose propionate is clear, amorphous, ridgid and dimensionally
stable. It has good resistance to gradient media including
Potassium Bromide and Sodium Chloride. It was used extensively
in the airfuge and Optima TL & TLX but has been discontinued.
Like all the amorphous materials it has poor chemical resistance
and is not suitable with alcohols, solvents, most acids and bases.
Sterilization method of choice is chemical. Recently it’s use has
been discontinued by Beckman.

• Advantages u Disadvantages
1. Clear 1. Poor chemical resistance
2. Strong 2. Not autoclavable
3. Inert
 Polyethylene
Polyethylene is translucent, stable and crystalline in structure. It
comes in many grades but those ridgid enough for tubes are
usually UHMWPE (Ultra-high molecular weight polyethylene). It
has excellent chemical resistance to most density media, acids,
bases, and alcohols. The use of Organic solvents are inappropriate.
Polyethylene can not be autoclaved.

• Advantages u Disadvantages
1. Great chemical 1. Not autoclavable
resistance 2. Not clear
2. Inert 3. Limited heat range
 Stainless Steel
Stainless steel tubes are very strong, chemically resistant, &
opaque. It is used in situations where plastic tubes can’t
withstand the centrifugation media. It is susceptible to corrosion,
if scratched or damaged and left in contact with salts. All
sterilization methods are suitable with stainless steel but they
should be thoroughly dried after autoclaving. Because of their
strength they can be run at any fill level. Because of their density
they require special speed reductions (information on special
considerations for stainless steel tubes is found in L5-TB-072C).

• Advantages u Disadvantages

1. Run at any fill level 1. Opaque

2. Chemically resistant 2. Not recommended with
3. Durable salt solutions
 Glass Tubes & Bottles
Glass tubes for centrifugation are produced primarily by Corning (Pyrex &
Corex) and Kimble (Kimax). Many blood drawing tubes such as Becton
Dickinsons Vacutainers are also glass. Because of it’s brittle nature glass
tubes are limited to low speeds (see PL 367 for speed limits for glass tubes in
Beckman rotors). It has excellent chemical resistance and can be sterilized
by all of the common means. Scratches or chips weaken glass and freezing
of samples is not recommended. These are wettable, which means fluids will
not form drops on the surface which is important to some researchers who
hand layer gradients. Beckman no longer sells the Corex & Pyrex tubes but
does support their use in our rotors with adapters.

• Advantages u Disadvantages

1. Clear 1. Brittle
2. Wettable 2. Low speeds only
3. Chemically resistant
4. Wide temperature and
pH range
 Tube Life Expectancy
• It is important to note that all materials discussed so far have many
common factors that influence the life expectancy. Many are interrelated
and work together to shorten the life of the tube or bottles. Some of the
factors are:
Frequency of use affects product life
Maximum g-force exposure
Method of storing tubes, scratches, scuffs, chipping or exposure to
extremes of UV can shorten tube life expectancy
Method of cleaning or sterilization-- ex is autoclaving PC reduces
life up to 90%
Tube contents-solvents, ETOH etc can reduce life as can length of cycle.
Ex: four forty hour runs can be harder on the tube than 40 one hour runs
Combinations of these factors work together to shorten the tube life.
Many tubes are not reusable such as Quick Seals and OptSeals.
 Tube and Bottle Designs
• There are many different tube and bottle designs. The design is
dictated by the application. Design considerations include speed
or force, rotor design (SW, FA, VTi, NVT, ultra, high speed etc),
need for clarity and ease of use.
• Tremendous forces of ultracentrifugation require extremely tight tolerances.
The tube requires complete support which is provided by the rotor cell and the
fluid pressure of the sample. This is the reason we have tube spacers and rotor
plugs for the VTi and NVT rotors. Lets detail the differences between them
• SW--easiest style, open top tubes are supported by fluid
pressure, and with thick wall tubes can often run at
less than full fill levels
• FA--open top tubes not suitable except in very small tubes
tubes with cap assemblies, QS & OS use spacers to
provide closed system.
• VTi & NVT--very similar to FA but must have rotor
plug to counter hydrostatic pressure
 Open Top Tubes
• As name suggests they have straight side walls and either round (hemispherical)
or conical bottoms.
• Available in either thin wall or thick wall, in PC, PA or UC materials.
• Thin wall tubes are approximately .5mm thick. In SW are run W/O cap and fill
level is 2-3 mm from top. In FA rotors must use a cap assembly and be
completely full so that fluid pressure inside tubes supports the tube sidewall. Are
very easy to puncture or slice (this is how bands are removed typically).
Typically single use but with careful attention can be used a few times.
• Thick wall tubes are approximately 1-1.5mm thick. Walls can
support themselves, PC in particular. With exception of
TL 100 tubes speeds must be reduced when running at less
than full fill levels. It is always best to caution that we
can’t test at every speed and fill level so advise customer
to try run with dummy sample. Tubes are not puncturable
but PP and PA can easily be sliced. PC ruins blades on
slicer quickly. Thick wall tubes are durable and reusable.
 Bottles
• We distinguish bottles by the fact that they have threaded tops which accept a
cap. Bottles are thick wall vessels and are sold in polyallomer, polypropylene
and polycarbonate. They can have flat bottoms (large bottles such as 1L .5L,
.25L), round bottoms or conical bottoms . A common bottle style is called
Oakridge named after the Oakridge National Laboratories in Tennessee. The
Oakridge style bottles cap diameter is equal to the bottle diameter and usually
have rounded shoulders. Bottles must be at least half full, but again, unless
specifically stated in the rotor or tube manuals do not guarantee that a
particular fill level will work. Use a dummy sample to test the bottle
at the researchers conditions.
• Care should be taken when cleaning bottles. Scratches act as stress
concentrators and reduce the bottles life significantly (particularly with PC
tubes). Mild soap solutions are recommend and PA and PP can be
autoclaved. Never autoclave a bottle with the cap on. The pressure difference
between the autoclave and bottles collapses the bottle. PC should not be
autoclaved in order to maximize life expectancy.
 Caps
• There are several different kinds of caps available. Caps for ultracentrifuges
are made of harder materials to withstand the forces of centrifugation.
Materials such as aluminum, titanium, Delrin and Noryl are used. In lower
speed applications the caps are usually polypropylene or polyethylene as
they do not have to provide the support or withstand the tremendous forces
of the ultracentrifuge.

• Tube caps often outlast the bottle and so the bottles can be purchased
separately as can the cap assemblies and the individual parts of the
assembly.

• Just as with rotors, care should be taken when cleaning aluminum caps.
Damage to the anodized cap surfaces can allow corrosion which when
coupled with the stress of centrifugation can cause failure. A cap
centrifuged into a rotor cell can be very difficult to remove. In a titanium
rotor it can be sent to Palo Alto for chemical removal. In aluminum rotors
they often must be machined out.
 Beckman Bottles
• Advantages
Wide variety of sizes and materials

Cap assemblies
Choice of styles (conical etc)

Assurance of proper fit in Beckman

rotors

Warranty maintained
Designed to meet customer
requirements
 Trouble Shooting Guide
• Leaking Quick-Seal
Ø Most common is leak from the seal. Check for fluid path through the stem.
Ø Pinch-off areas can also be a problem so check bottom and shoulder.
Ø Check unused tubes for brittleness
Ø Customer feedback forms are very important for correcting problems.

• Capped Tubes
Ø Caps not properly assembled. Any moisture on
o-ring will cause leak.
Ø Worn or brittle gasket or o-ring.
Ø Wrong cap for tube (blue cap on thinwall tube)
Mismatched parts of cap
 Troubleshooting Continued
• Uncapped Tubes
Ø Maximum fill volume either exceeded or minimum fill level not met

• Tubes/Bottles (leaking)
Ø Check quality of bottle and look for excessive crazing, scratches or other signs of
excessive use. Check o-ring for brittleness, make sure sealing area is clean& dry. Check
area on bottle where the o-ring seats for nicks and scratches which can cause a fluid path.
Was cap tightened properly? Tube fill level correct?
• Tubes/Bottles (splitting)
Ø Look at bottle to see if it shows excessive wear (remember, they have a limited life
expectancy). Is the bottle being run with the proper rotor or adapter ie is the tube/rotor fit
proper? Run temperature above or below the recommended levels, tubes become brittle
below 2 degrees C and stretch when run
over 25 degrees. Are parts undersized? Check unused bottles for fit.
• Capped Tubes (crazing)
Ø Is normal and acceptable as tube stretches to fill rotor cavity If excessive check the media
to make sure it is compatible with tube material, check tube fit, age of tubes and usage
 Troubleshooting continued
• Tube lip pulled away from cap
Ø Cap not properly torqued, seated. Moisture on o-ring or sealing surface. O-ring greased
before starting run (rotor & instrument o-rings get greased not tube rings)
• Tube/Bottles swell
Ø Inappropriate solution used which is absorbed by tube material. Autoclaving with cap on.
Parts spun above recommended temperature.
Failure Modes
 Gradient Media
• Ultracentrifuges exist to separate particles. They do this by variations on two
themes. Isopycnic (same density) is used for banding and differential floatation
such as with lipoprotein work. Rate zonal work is done extensively and includes
the simplest separation-pelleting. Rate zonal is typically done in SW rotors so you
can see it is used commonly.
• Particles move in solution according to their buoyant density and size. Different
media is used according to the density of the particles of interest. Lets look at a
common separation, nucleic acids. Plasmid DNA and cDNA have a density of
about 1.55g/mL. CsCl is used as a media because it is a self forming gradient, ie it
forms a gradient from 1.0 to about 1.9 g/mL in the tube when centrifuged. The
nucleic acid then moves to the spot in the tube where it’s density equals the
density of the gradient (isopycnic) Care is taken not to overspeed CsCl gradients
because a saturated solution can precipitate out in a pellet (rotors are rated to
handle 1.2 to 1.7 g/mL and thus the CsCl precipitation curves in the manuals). The
pellet is pure CsCl and has a density of 4.0 g/mL which can damage or destroy a
rotor. Gradients are chosen for their gradient ranges and any effects they may
have on biological materials.
 Sucrose/Cesium Sulfate/Misc Media
• Sucrose is used commonly for rate zonal separations. It is not a self forming
gradient and is generally hand layered. The gradient is created by varying the
concentration of the sugar. Sucrose is quite common in separations where salt ions
in solution will interact with or bind the biological sample. Also useful for recycling
as coloring and carbonation is added and sold as soft drink.
• Cesium Sulfate, CsSO4 , is a salt and like CsCl is a self-forming gradient. It is often
used in RNA work as it has a “steeper” gradient than CsCl.
• NaCl is less dense than CsCl and primarily used for lipoprotein work and can be
used to season meat and poultry!
• Sodium Dodecyl Sulfate (SDS) is an ionic detergent often used in cell lysis (blowing
up a cell, destroying the cell wall to get to the contents) and to solubilize proteins
(dissolve in solution).
• Triton x-100, a non-ionic detergent used in cell lysis and solubilizing protein Also
used to aid in releasing RNA pellets from tube side wall in NVT rotors
• Ethidium Bromide (EtBr) is a fluorescent intercalating dye use in DNA preps to
separate cDNA and plasmid DNA and to aid in visualization of the band.
 Sterilization
• Beckman does not provide sterile centrifuge tubes so your customer will use many
techniques to sterilize them. Here is a sampling.
• Autoclaving--Done under pressure usually, uses heat to kill or denature biological
material harmful to sample such as RNAse, Protienases, etc. Make sure that
capped tubes are never autoclaved with the caps on. Typical conditions are 121
degrees Centigrade at 15 PSI of pressure. Often is used to extract stuck tubes
from rotors after tube failures by using a difference in the coefficient of thermal
expansion. UC can not be autoclaved and PC is damaged by autoclaving
• Ethylene Oxide (EtO) is a poisonous gas . It’s not common as it requires expensive
containment equipment & is regulated because of the danger.
• Chemical or cold sterilization is used by immersing tubes in chemical solutions to
sterilize. Common solutions are alcohols, hydrogen peroxide, and specially
formulated solutions. Consult manuals for information on compatibility of the
solution with tubes.
• Ultra Violet light is used to sterilize tubes, but it is a lengthy and expensive means
of sterilizing and not widely used. UV Harms many polymers !
• Irradiation is often used by manufacturers but because of expense seldom used in
labs. Can damage or discolor many polymers !
 Cap Types
• You will run in to many different colored cap assemblies in the field. There is a
method to the madness & here is some of the scoop:

Ø Red anodized caps are used with thin wall tubes ( PA or UC)

Ø Blue anodized caps go with thick wall tubes (PA, PP or PC)

Ø Black anodized caps are for thick wall tubes used in the Ty 30 rotor (either PA, PP or
PC)

Ø Clear anodized caps are used with many thick and thin wall tubes in a variety of rotors.
These are thrown in to confuse you!

» In addition there are some special cap assemblies for special rotors.
Following is a sampling
 Special Caps
• A special crimp lock cap assembly is required for maximum rotor speed
in the Type 70Ti rotor. The 1 X 31/4 inch thinwall polyallomer tube is
crimped between the titanium crown and the aluminum stem. This cap
provides the reliable seal necessary for centrifugation at maximum
speed in the 70Ti. Instructions for assembling the tube and cap are
provided in the rotor manual. A special tool kit part number 338841 is
required.

• Titanium caps are use for cap assemblies for thinwall UC and PA for
max rotor speed in the Types 90Ti, 80Ti, 75Ti, and 70.1Ti. The cap
crown is specially machined to lock onto the cap stem. To ensure
proper compression of the o-ring these caps must be tightened with a
torque wrench while the capped tube is held in the tube-cap vise.
 Cap assemblies
• Most of the open top tubes Beckman sells are
used in ultracentrifuges . The caps are specially
made to withstand the incredible forces of
centrifugation. Several components make
up a cap assembly. These include a stem, a nylon
insert, o-ring or flat gasket, nut, crown, Delrin
crown washer, and a stainless steel set screw.
When the nut is tightened or torqued the o-ring is
compressed against the wall of the tube creating
the seal. The stem wall serves to support the tube
and therefore the stem is longer for thinwall tubes.
Some stems are grit blasted on the surface that
contacts the tube to increase the friction and keep
the stem from pulling away from the tube. The cap
rests in the counterbore of the tube cavity and is
slightly larger in diameter than the tube This keeps
the tube from bearing the entire weight of the cap
and tube contents at speed. Thin wall tubes and
capped tubes in general must be run completely
full
 Filling Capped Tubes
• How do you fill a tube completely then
put on a cap assembly without spilling
or getting moisture on the o-ring or o-
ring seating area?

• Most of the caps have a hole in the

middle of the cap for this purpose
which is sealed with a stainless steel
set screw after filling. We will practice
filling tubes and the technique for
sealing these tubes will be shown. It
is tedious and time consuming and for
inexperienced personnel leaks often
occurred. It will be a good reminder of
why Quick-Seal and OptiSeal tubes
were developed.
 Maintenance of Caps
• The ultracentrifuge cap assemblies are
reusable. They are generally quite
expensive and require careful
maintenance in order to maximize their
useful life.
• They should be kept clean and dry.
Beckman solution 555 is recommended
as it is of neutral pH and will not damage
anodized parts. Cotton swabs or
brushes that will not scratch the surface
should be used when cleaning the
assemblies
• All parts should be rinsed in distilled
water and dried thoroughly before storing
• Stem threads should be lubricated
periodically using the Spinkote lubricant
provided with the rotor.
• Remember to keep the o-ring free of
grease or moisture when sealing tubes,
inspect components often, and never,
ever mix cap components
 Replacing Nylon Inserts
• The Nylon inserts fit into the cap stem and
can become worn or loose with continued
use. If the insert is worn the setscrew will
not seal the cap and it should be replaced
• To replace:
1. Remove the set screw & firmly fit
the Nylon insert tool PN 302460
into the stem

2. Unscrew the insert

3. Fit a new insert PN 302312 on the

end of the tool and screw it into the
stem until it bottoms firmly against
the stem threads
 Tightening Caps
• Tighten the caps by
Ø Use a Beckman torque wrench with
scale in inch pounds.
Ø For titanium caps 341968, 90 to 100
inch pounds or 10 to 11 N-M
Ø For red, blue and black anodized
aluminum caps 100 to 120 in
pounds or 10-1.6 N-M should be
used for the first four runs and then
100 inch pounds 11N-M thereafter
Ø For cap 338903 used in the Type 21
rotor use 100 inch pounds (11 N-M)
 Filling Level Chart
• Open Top tubes without caps
This chart is used to determine the
maximum fill level in thick wall open
top tubes when run without a cap. It
is not applicable when the tubes are
run in a rotor with a larger diameter
than the tube (using an adapter). The
same information is usually found in
the fine print of the product selection
guide. When using small tubes with
adapters the fill level must be
determined empirically.
 Quick-Seal Tubes
• Features u Benefits

Ø Thin wall design – Desired particles can be obtained

by puncturing or slicing
Ø Unique Quick-Seal Design – Worry free secure closure

Ø Unique Heat Sealing system – Secure closure without a cap

Ø Choice of PA or UC – Material to meet customers

specific needs
Ø Wide line of spacers – Permits use of many sized tubes in
Beckman rotors. Can use in all
vertical rotors, fixed angle or
swinging bucket
 Filling Quick-Seal Tubes
• Base of seal former at
topline on the neck of
tube and liquid meniscus
between the top and
bottom lines on neck
• Assure that fluid level is
correct and not in stem.
Moisture in stem will be
vaporized and can form
pockets in the stem which
can provide a fluid path
and cause a leak.
• Make sure seal former is
not depressed too far
(down onto the body of
the tube
 G-Max Tubes
• By using patented Quick-Seal tubes and floating spacers Beckman is able to allow
the use of small volume tubes while maintaining the maximum force on the sample.
The term floating spacer refers to the spacers ability to move in the centrifuge bore.
Spacers for most full size tubes sit on the counterbore of the rotor while floating
spacers can sit anywhere within the cell hole. By shortening the path length the
particles must travel (and not reducing force) run times can be significantly, often 1/3
to 1/4 the time required with full size tubes. Also useful where the amount of
sample does not match the full volume of the rotor.
 Konical Tubes
• Konical tubes come either
as open top or Quick-Seal
in Polyallomer only.

• Primary usage is for

concentrating pellets in
tube bottom of tube.

• Used in SW rotors

• Optimizes pelleting time

 OptiSeal tubes
• Designed for Fixed angle, Vertical and Near Vertical rotors.

• Example of the “clear” polyolefin materials. It is a

copolymer of propylene and ethylene which is roughly
equivalent to polyallomer (remember, polyallomer is a trade
name).

• No need for sealing devices or tube caps

• Tube seal is made by the fluid pressure in the tube and the
seal is made stronger as the g-force increases

• Spacers are required for all OptiSeal tubes

• Tube rack is essential for sealing the OptiSeal tubes for

use in fixed angle rotors. Holding tube to fill can lead to
leaks.
 OptiSeal Tubes
• Temperature range from 2-25 degrees
Celsius
• Autoclavable (121 degrees C for
approximately 30 minutes
• Tube plugs and tubes are single
usage only. Box of 56 tubes comes
with 56 plugs. There is no part
number set up for plugs so if they are
lost they can re-use the plugs they do
have. While not recommended, I have
tested the plugs in multiple runs
• Plugs , spacers and racks are
autoclavable
• Regular and Bell Top versions are
available. Bell tops are used in the
fixed angle rotors
 Filling OptiSeal Tubes
• Consult instructions with the tubes for
detailed instructions.
• Opening is large enough for volumetric
pipette.
• Be absolutely sure that there is no fluid in
the stem or on the plug or plug o-ring.
Fluid here provides a fluid path and a
small leak can occur which will usually
clear itself as hydrostatic pressure builds
up in tube
• The 3.3 mL OptiSeal tube PN 361627
and the 9.8 mL tube PN 361629 have
spacers that snap on. To avoid
disturbing sample place spacers on tube
before filling
 Temperature & Chemical
Resistance's
• Beckman tube materials are tested between 2 and 25 degrees Celsius. The tubes
go through what is called a “glass transition” between 0 and 2 degrees and become
brittle. While they are suitable in most cases for storing material at lower
temperatures the brittleness causes the tubes to fail if centrifuged. Be sure the tube
has thawed above 2 degrees if the tube was used for freezing a sample before
processing. I always suggest the customer pretest the procedure with a water
sample if they need to run at temperatures between 0-2 degrees.
• Chemical resistance's can be found in IN-175 (which is in the Black spiral bound
handout Rotors and Tubes for preparative ultracentrifuges. Most of this
information is generated by the manufacturers of the polymers and is given for
materials in static conditions (no loads). The highly dynamic loads of centrifugation
add another factor to the chemical resistance data. Again, it is usually best to
pretest the tubes before trying the run with a valuable sample. Of note is that phenol
and chloroform show as not marginal to unsatisfactory in the chart but are used
extensively with polyallomer and polypropylene.
•
Adapters
Spacers sit on top of the tube
and Spacers
and are designed to counter
the fluid pressure inside the
tube. Floating spacers sit in
cell bore while spacers typically
sit on counterbore of rotor
• Adapters are used to run tubes
with diameters smaller than the
rotor cell bore diameter, or
when using a conical shape
tube in a rotor with a
hemispherical bottom. Often
used when customers need to
use a smaller volume than the
rotor used normally
accommodates
• Adapters reduce the rmax of the
rotor, thus the g-force & k
factor. Beckman g-max tubes
do not!
 Adapter Materials
• Ultracentrifuge adapters are almost always made of
Delrin. Delrin has a density of 1.4 g/mL so rotor
speeds often must be derated. Delrin is typically
white and feels slick to the touch. It is not
mechanically strong so must be supported by tube
and tube contents.
• Noryl is sometimes used, and is more ridgid. Noryl is
black usually
• Delrin and Noryl can be autoclaved
• Lower speed adapters are often made of a filled
polypropylene and can be autoclaved
 Spacer Materials
• Spacers for ultracentrifuge tubes are usually anodized
aluminum, Noryl, Delrin, or Ultem (polyetherimide) an
amber, very strong but expensive material
• Spacers are all autoclavable
• Counter hydrostatic pressure. Pressure will burst tube if
spacer is not used.
• Floating spacers are patented and thus provide a lock out
spec for Beckman
• Spacers must be carefully maintained to avoid premature
failure. They are very long lived when maintained properly
Spacers
 Removing Stuck Tubes
• Stuck tubes, bottles and tube caps are a common problem in customer labs.
Many customers use a screw driver or other sharp instrument to remove the stuck
parts, often damaging the rotor. To prevent unnecessary rotor damage, try the
following to remover stuck plastic tubes & bottles
Ø Remove the sample if possible
Ø Place the rotor upside down in the autoclave and run at 121 degrees C for 15-30
minutes
Ø Do not autoclave samples that are incompatible with the heating cycle

This procedure also works for stuck spacers and caps. The different coefficients of
thermal expansion of the materials allows removal in most cases. When a tube cap fails
and centrifuges into the cavity it can be removed chemically if aluminum. If it is stainless
steel or titanium it must be returned to the factor and we will attempt to machine it out.
Often the rotor will have been damaged by the cap centrifuging into the cavity and will
not be repairable
 Quick Seal Tubes as
Freezing Vials
• An ideal marketing opportunity for any product is to find a new market or
application for an existing product. Investment in development is minimal and it
creates an opportunity for sales growth. Such is the case with using Quick Seal
tubes as freezing vials.
• Both Ultra-Clear and Polyallomer have been tested and found to perform
satisfactorily for freezing samples at temperatures down to -60 degrees. The
samples were tested in sealed tubes filled approximately 2/3 full for five days.
The tubes survived with no problems
• Remember the tubes must not be filled completely full to allow for expansion. 2/3
full seems to be an ideal fill
level. Try suggesting this new use in a few
labs and see what happens. The tubes are
convenient as they are already in the lab
most of the time and they will not leak
 Accessories
• Beckman offers
numerous
accessories for it’s
tubes and bottles.
The best way to
learn them is to view
them and ideally to
get hands on use.
Here are a few of
the items we offer.
 Accessories

• Tube
removal
tools
 Accessories
• Tube
Toppers
Method A
Method B
 Accessories

• Fraction
Recovery
System
 Chi Contattare
• Per informazioni tecniche-scientifiche
Marco Chiereghin
[email protected]
02.95392350

• Per richiesta di Intervento

199.184.184