Centrifuge and Its Types

Centrifuge and their types
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, the viscosity of the medium and rotor speed.
Principle
The centrifuge works using the sedimentation principle, where the
centripetal acceleration causes denser substances and particles to
move outward in the radial direction.
In general, applications for centrifugation
specify the degree of acceleration to be applied to
the sample rather than specifying a specific g Force (RCF) = (rpm)2 × 1.118 × 10-
rotational speed such as revolutions per minute. 5
× r
The acceleration is typically given in gravity [× RPM = √[RCF/(r × 1.118)] × 1 × 105
g] (or multiples of x g or g-force), the standard g = Relative Centrifuge Force
acceleration value due to gravity at the Earth’s r = rotational radius (cm)
surface (9.81 m/s2). The distinction between rpm N = Revolutions Per Minute (RPM)
and rcf is important, as two rotors with different
diameters running at the same rotational speed
(rpm) will result in different accelerations (rcf)
Types of Centrifugation
Preparative centrifugation: It is a technique concentrating

one component from the other.
Analytical centrifugation: It is a method that measures the
physical, chemical and hydrodynamic properties of the
suspended molecules.
1.Density Gradient Centrifugation

2.Differential centrifugation
1. Rate-Zonal Density-Gradient Centrifugation
2. Isopynic Centrifugation
Differential Centrifugation
• It is the most common type of centrifugation employed.
• Tissue such as the liver is homogenized at 32 degrees in a sucrose
solution that contains buffer.
• The homogenate is then placed in a centrifuge and spun at
constant centrifugal force at a constant temperature.
• After some time a sediment forms at the bottom of a centrifuge
called pellet and an overlying solution called supernatant.
• The overlying solution is then placed in another centrifuge tube
which is then rotated at higher speeds in progressing steps.
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 ultracentrifuges.
• The particles travel through the gradient until they reach a point at
which their density matches the density of surrounding sucrose.
• The fraction is removed and analyzed.
Rate-Zonal Density-Gradient Centrifugation
• Zonal centrifugation is also known as band or
gradient centrifugation
• It relies on the concept of sedimentation
coefficient (i.e. movement of sediment through
the liquid medium)
• In this technique, a density gradient is created in a
test tube with sucrose and high density at the
bottom.
• The sample of protein is placed on the top of the
gradient and then centrifuged.
• With centrifugation, faster-sedimenting particles in
sample move ahead of slower ones i.e. sample
separated as zones in the gradient.
• The protein sediment according to their
sedimentation coefficient and the fractions are
collected by creating a hole at the bottom of the
Isopynic Centrifugation
• The sample is loaded into the tube with the gradient-forming

solution (on top of or below pre-formed gradient, or mixed in
with self-forming gradient)
• The solution of the biological sample and cesium salt is uniformly
distributed in a centrifuge tube and rotated in an ultracentrifuge.
• Under the influence of centrifugal force, the cesium salts
redistribute to form a density gradient from top to bottom.
• Particles move to point where their buoyant density equals that
part of gradient and form bands. This is to say the sample
molecules move to the region where their density equals the
density of gradient.
• It is a “true” equilibrium procedure since depends on bouyant
densities, not velocities
Applications of Centrifugation
•To separate two miscible substances
•To analyze the hydrodynamic properties of macromolecules
•Purification of mammalian cells
•Fractionation of subcellular organelles (including membranes/membrane
fractions) Fractionation of membrane vesicles
•Separating chalk powder from water
•Removing fat from milk to produce skimmed milk
•Separating particles from an air-flow using cyclonic separation
•The clarification and stabilization of wine
•Separation of urine components and blood components in forensic and
research laboratories
•Aids in the separation of proteins using purification techniques such as
salting out, e.g. ammonium sulfate precipitation.

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Centrifuge and their types

Centrifugation is a technique of separating substances which involves the application of

Preparative centrifugation: It is a technique concentrating

1.Density Gradient Centrifugation

• The sample is loaded into the tube with the gradient-forming

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