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Presented by,
Maria Babu
S4 botany
CENTRIFUGATI
ON
Basic principle of Centrifugation

◦ When an object is moved in a circle with a steady angular velocity it will experience an inwardly
directed force F
◦ Angular velocity is the rate of velocity at which an object or a particle is rotating around a centre or a
specific point in a given time period
◦ The magnitude of F is determined by the angular velocity (w) and the radius of rotation (r) and is
represented as
During centrifugation, the suspended particles settle down on the bottom
of the centrifuge tube due to gravity .
◦ the suspension left above the sediments after centrifugation is called
supernatant.
◦ it may be subjected to repeated centrifugation, with progressively
increasing speed.
◦ As a result different particles settle down differently at different times ,at
different rates and at different sites .
◦ this may be called differential centrifugation .
◦ the supernatant left after the final centrifugation is called soluble fraction.
• Centrifuge roters are designed to generate
rotation speed that can bring about the
separation of the component in a sample.

• these are the three main types of roters

used in centrifuge.
1.Fixed angle rotors
◦ These roters hold the sample tubes at an angle of 45°
in relation to the axis of rotor

◦ In this type of rotor,the particles strike the opposite

side of the tube where the particles finally side down
and are collected at the bottom.

◦ These are faster than other types of rotors.however,as

the direction of the force is different from the position
of the tube,some particles might remain at the sides of
the tubes.
 2.Swinging bucket rotors/ Horizontal
rotors
◦ Swinging bucket rotors hold the tubes at an angle
of 90° or in a horizontal position as the rotor
swings .
◦ In this type of rotors,the particles are present along
the direction or the path of the force that allows the
particles to be moved away from the rotor towards
the bottom of the tubes.
◦ Because the tubes remains as a flat surface
allowing the deposited particles to be separated
from the supernatant.
 3.Vertical rotors
◦ Vertical rotors provide the shortest path
length,fastest run time, and the highest resolution of
all the rotors.
◦ In vertical rotors, the tubes are vertical during
operation of centrifuge.
◦ The yeild of the rotor is not as ideal as the position
of the tube doesn’t align with the direction of the
centrifugal force.
◦ As a result,instead of settling down, particles tend
to spread towards the outer wall of the tubes .
◦ These are isopycnic and density gradient
centrifugation.