APPLICATIONS

OF CENTRIFUGE
There are two techniques on
which the application of
centrifuge is based on:
PREPARATIVE
TECHNIQUE 1
• Preparative centrifugation is used for separating particles
according to their densities, isolating and/or harvesting
denser particles for collection in the pellet, and clarifying
suspensions containing particles. Sometimes researchers
also use preparative ultracentrifuges if they need the
flexibility to change the type of rotor in the instrument.
Preparative ultracentrifuges can be equipped with a wide
range of different rotor types, which can spin samples of
different numbers, at different angles, and at different
speeds
 “
• VELOCITY SEDIMENTATION
CENTRIFUGATION
• PREPARATIVE DIFFERENTIAL
CENTRIFUGATION
ANALYTICAL
TECHNIQUE 2
• Analytical centrifugation incorporate a scanning
visible/ultraviolet light-based optical detection system for
real-time monitoring of the sample’s progress during a spin.
This lets you watch the sedimentation process (or rather, a
detector collects light absorption data, which a computer
digitizes and records for you), allowing you to see the sample
concentrate as the centrifugal force increases.
• Analytical centrifugation can yield important information
about molecules in your sample, such as their overall shape,
conformational changes, and even the number and
stoichiometry of subunits that comprise protein complexes.
• All analytical techniques require use of an ultra centrifuge
and can be classified as
1. DIFFERENTIAL CENTRIFUGATION
2. DENSITY GRADIENT CENTRIFUGATION
A)ZONAL CENTRIFUGATION
B)ISOPYCNIC CENTRIFUGATION
 DIFFERENTIAL CENTRIFUGATION

Differential centrifugation is a technique commonly used by

biochemists. It is a common procedure in
microbiology and cytology used to separate
certain organelles from whole cells for further analysis of
specific parts of cells. In the process, a tissue sample is
first lysed to break the cell membranes and mix up the cell
contents.
 DIFFERENTIAL CENTRIFUGATION
• tissue such as liver is
homogenized at 32 degree
in a sucrose solution that
contains buffer
• the homogenate is then
placed in a centrifuge and
spun at constant
centrifugal force at
constant temperature.
• after sometime a sediment
forms at the bottom of
centrifuge called pellet
and overlying solution
called supernatant.
• the overlying solution is
then placed in another
centrifuge tube which is
then rotated at higher
speeds.
 DENSITY GRADIENT CENTRIFUGATION
Density- gradient centrifugation is a widely-used centrifugation
technique used to create relatively pure fractions of cellular
components. In fact, density-gradient centrifugation is capable of
creating purer fractions than is differential centrifugation.

Density-gradient centrifugation involves the centrifugation of

material through a centrifugation medium of higher or graded
density. When exposed to a elevated centrifugal force, cellular
components migrate through the medium and separate based on
their density.
 ZONAL CENTRIFUGATION
Zonal centrifugation relies on the concept of sedimentation
coefficient (ie movement of sediment through 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.
The proteins sediment according to their sedimentation coefficient
and the fractions are collected by creating a hole at the bottom of
tube.
 ISOPYCNIC CENTRIFUGATION

Isopycnic centrifugation is a method where the components of a

sample (e.g. DNA) are separated on the basis of their density in a
centrifuge according to the centrifugal force they experience. A
density gradient is established via equilibrium sedimentation (e.g.
using caesium ions), and then analyte molecules concentrate as
bands where the molecule density matches that of the surrounding
solution.