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Centrifugation

The document discusses different types of centrifuges including low speed, high speed, and ultracentrifuges. It describes their components and uses, such as separating substances by size, shape, density or viscosity. Conversion between RPM and RCF is also covered. Types of rotors and centrifugation techniques like differential pelleting and density gradient centrifugation are outlined.

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Muhammad Asif Shaheen
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340 views

Centrifugation

The document discusses different types of centrifuges including low speed, high speed, and ultracentrifuges. It describes their components and uses, such as separating substances by size, shape, density or viscosity. Conversion between RPM and RCF is also covered. Types of rotors and centrifugation techniques like differential pelleting and density gradient centrifugation are outlined.

Uploaded by

Muhammad Asif Shaheen
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as PPTX, PDF, TXT or read online on Scribd
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Centrifugation

Muhammad Asif Shaheen


Lecturer Pathology
Department of Pathology
KEMU, Lahore.
• Centrifugation is a technique of separating substances which involves
the application of centrifugal force.
• The particles are separated from a solution according to their
• Size
• Shape
• Density
• viscosity of the medium and
• rotor diamter.
Types
• LOW SPEED CENTRIFUGE
• The low speed centrifuge has a maximum speed of 4000-5000rpm
• These instruments usually operate at room temperatures with no
means of temperature control
• HIGH SPEED CENTRIFUGES
• High speed centrifuges are used in more sophisticated biochemical
applications, higher speeds and temperature control of the rotor
chamber are essential.
• High speed centrifuge has a speed of 6000-30000 RPM
• The operator of this instrument can carefully control speed and
temperature which is required for sensitive biological samples.
• ULTRACENTRIFUGES
• It is the most sophisticated instrument.
• Ultracentrifuge has a maximum speed of 150000 RPM
• Intense heat is generated due to high speed thus the spinning
chambers must be refrigerated and kept at high vacuum.
• It is used for both preparative work and analytical work
• Airfuge™ Air-Driven Ultracentrifuges from Beckman Coulter Life
Sciences
• With the ability to provide a simple, rapid method for removing
chylomicrons (fat particles in lipemic samples), the Airfuge Air-Driven
Ultracentrifuge delivers accurate results for lipemic clarification.
Lipemic samples can be cleaned in 10 minutes, eliminating the need
for sendouts.
• Speed of 80000 rpm
RPM and RCF
• Revolutions Per Minute (RPM)
• Revolutions Per Minute (RPM) in regards to centrifugation is simply a
measurement of how fast the centrifuge rotor does a full rotation in
one minute.
• Relative Centrifugal Force (RCF)
• Relative Centrifugal Force (RCF) or g-force (both are the same, RCF is
expressed as units of gravity) is a measurement of the gravitational force
that a sample is subject to.
• The force is generated from the spinning of the rotor which, in turn, exerts
this force outward on the centrifuge tube.
• Not only does RCF take into account the speed of rotation, it also measures
the distance from the centre of rotation to give us a g-force measurement.
• RCF is the preferred method of measurement as it will remain the same
even if you are using a different centrifuge with a different rotor size.
http://insilico.ehu.es/mini_tools/rcf_rpm.php
Online convertors for RPM to RCF and vice
versa
• http://insilico.ehu.es/mini_tools/rcf_rpm.php
• https://www.fishersci.co.uk/gb/en/scientific-products/centrifuge-
guide/centrifuge-applications-tools/rpm-rcf-calculator.html
Types of rotors in centrifuges
Swinging bucket Fixed bucket
Types of Centrifugation

• Differential Pelleting (differential centrifugation)


• Particles of different densities or sizes in a suspension will sediment at
different rates, with the larger and denser particles sedimenting
faster. These sedimentation rates can be increased by using
centrifugal force. A suspension of cells subjected to a series of
increasing centrifugal force cycles will yield a series of pellets
containing cells of decreasing sedimentation rate.
• Density Gradient Centrifugation
• This type of centrifugation is mainly used to purify viruses, ribosomes,
membranes etc.
• A sucrose density gradient is created by gently overlaying lower
concentrations of sucrose on higher concentrations in centrifuge tubes
• The particles of interest are placed on top of the gradient and centrifuge in
ultra centrifuges.
• The particles travel through the gradient until they reach a point at which
their density matches with the density of surrounding sucrose.
• The the fraction is removed and analyzed.
Safety consideration
• Lid off
• Balance
• Manual manipulation
• Breakage
• Solid surface
•Thanks

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