SlideShare a Scribd company logo
CENTRIFUGATION
Technique in Biochemistry
 A centrifuge is a device for separating particles from a
solution according to their size, shape, density, viscosity
of the medium and rotor speed.
 Action of centrifugal force is used to promote accelerated
settling of particles in a solid-liquid mixture.
 Biological centrifugation is a process that uses centrifugal
force to separate and purify mixtures of biological
particles in a liquid medium. It is a key technique for
isolating and analysing cells, subcellular fractions,
supramolecular complexes and isolated macromolecules
such as proteins or nucleicacid.
 In a solution, particles whose
density is higher than that of the
solvent sink (sediment), and
particles that are lighter than it
float to the top. The greater the
difference in density, the faster
they move. If there is no
difference in density (isopyknic
conditions), the particles stay
steady. To take advantage of
even tiny differences in density
to separate various particles in a
solution, gravity can be replaced
with the much more powerful
“centrifugal force” provided by
a centrifuge
PRINCIPLE OF CENTRIFUGATION
Basis of separation:
 -Size
 -Shape
 -Density
Methodology:
 -Utilizes density difference between the
particles/macromolecules and the medium in
which these are dispersed
 -Dispersed systems are subjected to artificially
induced gravitational fields
RELATIVE CENTRIFUGAL FORCE
 the ratio of the centrifugal acceleration at a
specified radius and the speed to the standard
acceleration of gravity
 RCF =42 (rev min -1 ) 2 r/ 3600 x981= G/g
 RCF units are therefore dimensionless
(denoting multiples of g) and revolutions per
minute are usually abbreviated as r.p.m.:
RCF=1.12 x 10-5 r.p.m2 r.
CENTRIFUGAL FIELD
GENERAL STEPS IN BIOCHEMICAL SEPARATION
SEPARATION OF MACROMOLECULES
DENSITIES OF BIOLOGICAL MATERIAL
CELL FRACTIONATION
CLINICAL LABORATORY USE OF CENTRIFUGATION
 Remove cellular components from blood to provide
cell free plasma or serum for analysis.
 Concentrate cellular elements and other components
of biological fluid for microscopic or chemical
analysis.
 Remove chemically precipitated protein from an
analytical specimen.
 Separate protein bound or antibody bound ligand
from free ligand in immunochemical assay.
 Extract solute in biological fluids from aqueous to
organic solvents.
 Separate lipids compounds such as chylomicrons
from other compounds in plasma or serum or
lipoprotein from one another.
Types of
Centrifuge
 Fixed Angle Rotor
 Swinging Buckcet Rotor
 Ultra Centrifugation
 Axial Centrifugation
FIXED ANGLE ROTOR SWINGING BUCKET ROTOR
 Sediment particles have only
short distance to travel before
pelleting at the angle of 25 or 40
degree.
 Shorter run time.
 The most widely used rotor type.
 This is used to separate nuclei,
mitochondria, nucleosome and
isopycnic separations. Isopycnic
separation is when biological
particle and surrounding medium
attain the equal level at the zero
sedimentation rate.
 Longer distance of travel may
allow better separation, such as
in density gradient centrifugation.
 The particles sediment uniformly
along the horizontal tube.
 Easier to withdraw supernatant
without disturbing pellet.
ULTRACENTRIFUGATION AXIAL CENTRIFUGATION
 A very high speed
centrifuge with fixed head
rotors.
 Most important application
is in separating
lipoproteins.
 This is available both in
analytical and preparative
models.
 Allows tubes to spun in
vertical orientation which is
used in traditional
centrifugation.
THANK YOU

More Related Content

What's hot

Biological databases
Biological databasesBiological databases
Biological databases
Malla Reddy College of Pharmacy
 
Clustal W - Multiple Sequence alignment
Clustal W - Multiple Sequence alignment   Clustal W - Multiple Sequence alignment
Clustal W - Multiple Sequence alignment
The Oxford College Engineering
 
Major biological nucleotide databases
Major biological nucleotide databasesMajor biological nucleotide databases
Major biological nucleotide databases
Vidya Kalaivani Rajkumar
 
protein sequence analysis
protein sequence analysisprotein sequence analysis
protein sequence analysis
RamikaSingla
 
String.pptx
String.pptxString.pptx
String.pptx
RitikaChoudhary57
 
DNA & RNA isolation
DNA & RNA isolationDNA & RNA isolation
DNA & RNA isolation
gptgeetika
 
Protein fold recognition and ab_initio modeling
Protein fold recognition and ab_initio modelingProtein fold recognition and ab_initio modeling
Protein fold recognition and ab_initio modeling
Bioinformatics and Computational Biosciences Branch
 
Ultra centrifugation
Ultra centrifugationUltra centrifugation
Ultra centrifugation
Rajpal Choudhary
 
ICAT Technique in Proteomics
ICAT Technique in ProteomicsICAT Technique in Proteomics
ICAT Technique in Proteomics
Bahauddin zakariya university,Multan
 
Distance measure between two biological sequences
Distance  measure between  two biological  sequences Distance  measure between  two biological  sequences
Distance measure between two biological sequences
ShwetA Kumari
 
Hydrophobic interaction chromatography (1)
Hydrophobic interaction chromatography (1)Hydrophobic interaction chromatography (1)
Hydrophobic interaction chromatography (1)Rakesh Kumar
 
Reverse phase chromatography
Reverse phase chromatographyReverse phase chromatography
Reverse phase chromatography
faixan_live
 
Cloning vectors
Cloning vectorsCloning vectors
Cloning vectors
NOMI KhanS
 
Immunoprecipitation
ImmunoprecipitationImmunoprecipitation
Immunoprecipitation
Masuma Sani
 
Peptide mapping
Peptide mappingPeptide mapping
Peptide mapping
Saklecha Prachi
 
Affinity chromatography
Affinity chromatography Affinity chromatography
Affinity chromatography
Mr. MOHD FAHAD
 
Two-dimensional polyacrylamide gel electrophoresis (2D-PAGE)
Two-dimensional polyacrylamide gel electrophoresis (2D-PAGE)Two-dimensional polyacrylamide gel electrophoresis (2D-PAGE)
Two-dimensional polyacrylamide gel electrophoresis (2D-PAGE)
Santosh Kumar Sahoo
 

What's hot (20)

2d Page
2d Page2d Page
2d Page
 
Biological databases
Biological databasesBiological databases
Biological databases
 
Clustal W - Multiple Sequence alignment
Clustal W - Multiple Sequence alignment   Clustal W - Multiple Sequence alignment
Clustal W - Multiple Sequence alignment
 
Major biological nucleotide databases
Major biological nucleotide databasesMajor biological nucleotide databases
Major biological nucleotide databases
 
protein sequence analysis
protein sequence analysisprotein sequence analysis
protein sequence analysis
 
String.pptx
String.pptxString.pptx
String.pptx
 
DNA & RNA isolation
DNA & RNA isolationDNA & RNA isolation
DNA & RNA isolation
 
Protein fold recognition and ab_initio modeling
Protein fold recognition and ab_initio modelingProtein fold recognition and ab_initio modeling
Protein fold recognition and ab_initio modeling
 
Ultra centrifugation
Ultra centrifugationUltra centrifugation
Ultra centrifugation
 
ICAT Technique in Proteomics
ICAT Technique in ProteomicsICAT Technique in Proteomics
ICAT Technique in Proteomics
 
Distance measure between two biological sequences
Distance  measure between  two biological  sequences Distance  measure between  two biological  sequences
Distance measure between two biological sequences
 
Hydrophobic interaction chromatography (1)
Hydrophobic interaction chromatography (1)Hydrophobic interaction chromatography (1)
Hydrophobic interaction chromatography (1)
 
Reverse phase chromatography
Reverse phase chromatographyReverse phase chromatography
Reverse phase chromatography
 
Cloning vectors
Cloning vectorsCloning vectors
Cloning vectors
 
Immunoprecipitation
ImmunoprecipitationImmunoprecipitation
Immunoprecipitation
 
Peptide mapping
Peptide mappingPeptide mapping
Peptide mapping
 
Affinity chromatography
Affinity chromatography Affinity chromatography
Affinity chromatography
 
Gel exclusion chromatography
Gel exclusion chromatographyGel exclusion chromatography
Gel exclusion chromatography
 
Two-dimensional polyacrylamide gel electrophoresis (2D-PAGE)
Two-dimensional polyacrylamide gel electrophoresis (2D-PAGE)Two-dimensional polyacrylamide gel electrophoresis (2D-PAGE)
Two-dimensional polyacrylamide gel electrophoresis (2D-PAGE)
 
centrifugation
centrifugationcentrifugation
centrifugation
 

Similar to Centrifugation- Techniques in Biochemistry

Centrifugation.pptx
Centrifugation.pptxCentrifugation.pptx
Centrifugation.pptx
Praveen Sahu
 
Centrifugation Introduction.
Centrifugation Introduction.Centrifugation Introduction.
Centrifugation Introduction.
TejasBhatia2
 
Centrifugation
CentrifugationCentrifugation
Centrifugation
Amit mishra
 
1957992 635169139917595000
1957992 6351691399175950001957992 635169139917595000
1957992 635169139917595000MANJU461988
 
Centrifugation principle and types by Dr. Anurag Yadav
Centrifugation principle and types by Dr. Anurag YadavCentrifugation principle and types by Dr. Anurag Yadav
Centrifugation principle and types by Dr. Anurag Yadav
Dr Anurag Yadav
 
Centrifugation by Dr. Anurag Yadav
Centrifugation by Dr. Anurag YadavCentrifugation by Dr. Anurag Yadav
Centrifugation by Dr. Anurag Yadav
Dr Anurag Yadav
 
Centrifugation amirtham
Centrifugation   amirthamCentrifugation   amirtham
Centrifugation amirtham
christanantony
 
62 bioanalatyical.pptx
62 bioanalatyical.pptx62 bioanalatyical.pptx
62 bioanalatyical.pptx
DevaprasadPanda
 
Common Approaches for Centrifugation in the Laboratory.pdf
Common Approaches for Centrifugation in the Laboratory.pdfCommon Approaches for Centrifugation in the Laboratory.pdf
Common Approaches for Centrifugation in the Laboratory.pdf
RWDLifeScience
 
Centrifugation lab practical 8.pptx
Centrifugation  lab practical 8.pptxCentrifugation  lab practical 8.pptx
Centrifugation lab practical 8.pptx
Dyarzrar
 
Principles and methods of.pdf
Principles and methods of.pdfPrinciples and methods of.pdf
Principles and methods of.pdf
NeoRedWolves
 
Centrifuge.pptx
Centrifuge.pptxCentrifuge.pptx
Centrifuge.pptx
saraso888
 
Centrifugation
CentrifugationCentrifugation
Centrifugation
karanthakor1
 
How Density can be a Property in Bioseparation....
How Density can be a Property in Bioseparation....How Density can be a Property in Bioseparation....
How Density can be a Property in Bioseparation....
Razib Datta Shubhra
 
Downstream processing
Downstream processingDownstream processing
Downstream processing
SunandaArya
 
Downstream processing - industrial microbiology
Downstream processing - industrial microbiology Downstream processing - industrial microbiology
Downstream processing - industrial microbiology
Kiran Kumar
 
lab centrifuge ppt
 lab centrifuge ppt lab centrifuge ppt
lab centrifuge ppt
sumitkumar579644
 
Prvn ppt
Prvn pptPrvn ppt
Prvn ppt
praveenpkd22
 
prvnppt-141230053156-conversion-gate02.pdf
prvnppt-141230053156-conversion-gate02.pdfprvnppt-141230053156-conversion-gate02.pdf
prvnppt-141230053156-conversion-gate02.pdf
DevPanda5
 

Similar to Centrifugation- Techniques in Biochemistry (20)

Centrifugation.pptx
Centrifugation.pptxCentrifugation.pptx
Centrifugation.pptx
 
Centrifugation Introduction.
Centrifugation Introduction.Centrifugation Introduction.
Centrifugation Introduction.
 
Centrifugation
CentrifugationCentrifugation
Centrifugation
 
1957992 635169139917595000
1957992 6351691399175950001957992 635169139917595000
1957992 635169139917595000
 
Centrifugation principle and types by Dr. Anurag Yadav
Centrifugation principle and types by Dr. Anurag YadavCentrifugation principle and types by Dr. Anurag Yadav
Centrifugation principle and types by Dr. Anurag Yadav
 
Centrifugation by Dr. Anurag Yadav
Centrifugation by Dr. Anurag YadavCentrifugation by Dr. Anurag Yadav
Centrifugation by Dr. Anurag Yadav
 
Centrifugation amirtham
Centrifugation   amirthamCentrifugation   amirtham
Centrifugation amirtham
 
62 bioanalatyical.pptx
62 bioanalatyical.pptx62 bioanalatyical.pptx
62 bioanalatyical.pptx
 
Common Approaches for Centrifugation in the Laboratory.pdf
Common Approaches for Centrifugation in the Laboratory.pdfCommon Approaches for Centrifugation in the Laboratory.pdf
Common Approaches for Centrifugation in the Laboratory.pdf
 
Centrifugation lab practical 8.pptx
Centrifugation  lab practical 8.pptxCentrifugation  lab practical 8.pptx
Centrifugation lab practical 8.pptx
 
Principles and methods of.pdf
Principles and methods of.pdfPrinciples and methods of.pdf
Principles and methods of.pdf
 
Centrifuge.pptx
Centrifuge.pptxCentrifuge.pptx
Centrifuge.pptx
 
Centrifugation
CentrifugationCentrifugation
Centrifugation
 
How Density can be a Property in Bioseparation....
How Density can be a Property in Bioseparation....How Density can be a Property in Bioseparation....
How Density can be a Property in Bioseparation....
 
Downstream processing
Downstream processingDownstream processing
Downstream processing
 
Downstream processing - industrial microbiology
Downstream processing - industrial microbiology Downstream processing - industrial microbiology
Downstream processing - industrial microbiology
 
Centrifuge_Report
Centrifuge_ReportCentrifuge_Report
Centrifuge_Report
 
lab centrifuge ppt
 lab centrifuge ppt lab centrifuge ppt
lab centrifuge ppt
 
Prvn ppt
Prvn pptPrvn ppt
Prvn ppt
 
prvnppt-141230053156-conversion-gate02.pdf
prvnppt-141230053156-conversion-gate02.pdfprvnppt-141230053156-conversion-gate02.pdf
prvnppt-141230053156-conversion-gate02.pdf
 

Recently uploaded

GBSN- Microbiology (Lab 3) Gram Staining
GBSN- Microbiology (Lab 3) Gram StainingGBSN- Microbiology (Lab 3) Gram Staining
GBSN- Microbiology (Lab 3) Gram Staining
Areesha Ahmad
 
Circulatory system_ Laplace law. Ohms law.reynaults law,baro-chemo-receptors-...
Circulatory system_ Laplace law. Ohms law.reynaults law,baro-chemo-receptors-...Circulatory system_ Laplace law. Ohms law.reynaults law,baro-chemo-receptors-...
Circulatory system_ Laplace law. Ohms law.reynaults law,baro-chemo-receptors-...
muralinath2
 
erythropoiesis-I_mechanism& clinical significance.pptx
erythropoiesis-I_mechanism& clinical significance.pptxerythropoiesis-I_mechanism& clinical significance.pptx
erythropoiesis-I_mechanism& clinical significance.pptx
muralinath2
 
Nutraceutical market, scope and growth: Herbal drug technology
Nutraceutical market, scope and growth: Herbal drug technologyNutraceutical market, scope and growth: Herbal drug technology
Nutraceutical market, scope and growth: Herbal drug technology
Lokesh Patil
 
Comparative structure of adrenal gland in vertebrates
Comparative structure of adrenal gland in vertebratesComparative structure of adrenal gland in vertebrates
Comparative structure of adrenal gland in vertebrates
sachin783648
 
Seminar of U.V. Spectroscopy by SAMIR PANDA
 Seminar of U.V. Spectroscopy by SAMIR PANDA Seminar of U.V. Spectroscopy by SAMIR PANDA
Seminar of U.V. Spectroscopy by SAMIR PANDA
SAMIR PANDA
 
THE IMPORTANCE OF MARTIAN ATMOSPHERE SAMPLE RETURN.
THE IMPORTANCE OF MARTIAN ATMOSPHERE SAMPLE RETURN.THE IMPORTANCE OF MARTIAN ATMOSPHERE SAMPLE RETURN.
THE IMPORTANCE OF MARTIAN ATMOSPHERE SAMPLE RETURN.
Sérgio Sacani
 
Structural Classification Of Protein (SCOP)
Structural Classification Of Protein  (SCOP)Structural Classification Of Protein  (SCOP)
Structural Classification Of Protein (SCOP)
aishnasrivastava
 
Earliest Galaxies in the JADES Origins Field: Luminosity Function and Cosmic ...
Earliest Galaxies in the JADES Origins Field: Luminosity Function and Cosmic ...Earliest Galaxies in the JADES Origins Field: Luminosity Function and Cosmic ...
Earliest Galaxies in the JADES Origins Field: Luminosity Function and Cosmic ...
Sérgio Sacani
 
Leaf Initiation, Growth and Differentiation.pdf
Leaf Initiation, Growth and Differentiation.pdfLeaf Initiation, Growth and Differentiation.pdf
Leaf Initiation, Growth and Differentiation.pdf
RenuJangid3
 
In silico drugs analogue design: novobiocin analogues.pptx
In silico drugs analogue design: novobiocin analogues.pptxIn silico drugs analogue design: novobiocin analogues.pptx
In silico drugs analogue design: novobiocin analogues.pptx
AlaminAfendy1
 
GBSN - Biochemistry (Unit 5) Chemistry of Lipids
GBSN - Biochemistry (Unit 5) Chemistry of LipidsGBSN - Biochemistry (Unit 5) Chemistry of Lipids
GBSN - Biochemistry (Unit 5) Chemistry of Lipids
Areesha Ahmad
 
in vitro propagation of plants lecture note.pptx
in vitro propagation of plants lecture note.pptxin vitro propagation of plants lecture note.pptx
in vitro propagation of plants lecture note.pptx
yusufzako14
 
What is greenhouse gasses and how many gasses are there to affect the Earth.
What is greenhouse gasses and how many gasses are there to affect the Earth.What is greenhouse gasses and how many gasses are there to affect the Earth.
What is greenhouse gasses and how many gasses are there to affect the Earth.
moosaasad1975
 
4. An Overview of Sugarcane White Leaf Disease in Vietnam.pdf
4. An Overview of Sugarcane White Leaf Disease in Vietnam.pdf4. An Overview of Sugarcane White Leaf Disease in Vietnam.pdf
4. An Overview of Sugarcane White Leaf Disease in Vietnam.pdf
ssuserbfdca9
 
Deep Behavioral Phenotyping in Systems Neuroscience for Functional Atlasing a...
Deep Behavioral Phenotyping in Systems Neuroscience for Functional Atlasing a...Deep Behavioral Phenotyping in Systems Neuroscience for Functional Atlasing a...
Deep Behavioral Phenotyping in Systems Neuroscience for Functional Atlasing a...
Ana Luísa Pinho
 
insect taxonomy importance systematics and classification
insect taxonomy importance systematics and classificationinsect taxonomy importance systematics and classification
insect taxonomy importance systematics and classification
anitaento25
 
Multi-source connectivity as the driver of solar wind variability in the heli...
Multi-source connectivity as the driver of solar wind variability in the heli...Multi-source connectivity as the driver of solar wind variability in the heli...
Multi-source connectivity as the driver of solar wind variability in the heli...
Sérgio Sacani
 
RNA INTERFERENCE: UNRAVELING GENETIC SILENCING
RNA INTERFERENCE: UNRAVELING GENETIC SILENCINGRNA INTERFERENCE: UNRAVELING GENETIC SILENCING
RNA INTERFERENCE: UNRAVELING GENETIC SILENCING
AADYARAJPANDEY1
 
Astronomy Update- Curiosity’s exploration of Mars _ Local Briefs _ leadertele...
Astronomy Update- Curiosity’s exploration of Mars _ Local Briefs _ leadertele...Astronomy Update- Curiosity’s exploration of Mars _ Local Briefs _ leadertele...
Astronomy Update- Curiosity’s exploration of Mars _ Local Briefs _ leadertele...
NathanBaughman3
 

Recently uploaded (20)

GBSN- Microbiology (Lab 3) Gram Staining
GBSN- Microbiology (Lab 3) Gram StainingGBSN- Microbiology (Lab 3) Gram Staining
GBSN- Microbiology (Lab 3) Gram Staining
 
Circulatory system_ Laplace law. Ohms law.reynaults law,baro-chemo-receptors-...
Circulatory system_ Laplace law. Ohms law.reynaults law,baro-chemo-receptors-...Circulatory system_ Laplace law. Ohms law.reynaults law,baro-chemo-receptors-...
Circulatory system_ Laplace law. Ohms law.reynaults law,baro-chemo-receptors-...
 
erythropoiesis-I_mechanism& clinical significance.pptx
erythropoiesis-I_mechanism& clinical significance.pptxerythropoiesis-I_mechanism& clinical significance.pptx
erythropoiesis-I_mechanism& clinical significance.pptx
 
Nutraceutical market, scope and growth: Herbal drug technology
Nutraceutical market, scope and growth: Herbal drug technologyNutraceutical market, scope and growth: Herbal drug technology
Nutraceutical market, scope and growth: Herbal drug technology
 
Comparative structure of adrenal gland in vertebrates
Comparative structure of adrenal gland in vertebratesComparative structure of adrenal gland in vertebrates
Comparative structure of adrenal gland in vertebrates
 
Seminar of U.V. Spectroscopy by SAMIR PANDA
 Seminar of U.V. Spectroscopy by SAMIR PANDA Seminar of U.V. Spectroscopy by SAMIR PANDA
Seminar of U.V. Spectroscopy by SAMIR PANDA
 
THE IMPORTANCE OF MARTIAN ATMOSPHERE SAMPLE RETURN.
THE IMPORTANCE OF MARTIAN ATMOSPHERE SAMPLE RETURN.THE IMPORTANCE OF MARTIAN ATMOSPHERE SAMPLE RETURN.
THE IMPORTANCE OF MARTIAN ATMOSPHERE SAMPLE RETURN.
 
Structural Classification Of Protein (SCOP)
Structural Classification Of Protein  (SCOP)Structural Classification Of Protein  (SCOP)
Structural Classification Of Protein (SCOP)
 
Earliest Galaxies in the JADES Origins Field: Luminosity Function and Cosmic ...
Earliest Galaxies in the JADES Origins Field: Luminosity Function and Cosmic ...Earliest Galaxies in the JADES Origins Field: Luminosity Function and Cosmic ...
Earliest Galaxies in the JADES Origins Field: Luminosity Function and Cosmic ...
 
Leaf Initiation, Growth and Differentiation.pdf
Leaf Initiation, Growth and Differentiation.pdfLeaf Initiation, Growth and Differentiation.pdf
Leaf Initiation, Growth and Differentiation.pdf
 
In silico drugs analogue design: novobiocin analogues.pptx
In silico drugs analogue design: novobiocin analogues.pptxIn silico drugs analogue design: novobiocin analogues.pptx
In silico drugs analogue design: novobiocin analogues.pptx
 
GBSN - Biochemistry (Unit 5) Chemistry of Lipids
GBSN - Biochemistry (Unit 5) Chemistry of LipidsGBSN - Biochemistry (Unit 5) Chemistry of Lipids
GBSN - Biochemistry (Unit 5) Chemistry of Lipids
 
in vitro propagation of plants lecture note.pptx
in vitro propagation of plants lecture note.pptxin vitro propagation of plants lecture note.pptx
in vitro propagation of plants lecture note.pptx
 
What is greenhouse gasses and how many gasses are there to affect the Earth.
What is greenhouse gasses and how many gasses are there to affect the Earth.What is greenhouse gasses and how many gasses are there to affect the Earth.
What is greenhouse gasses and how many gasses are there to affect the Earth.
 
4. An Overview of Sugarcane White Leaf Disease in Vietnam.pdf
4. An Overview of Sugarcane White Leaf Disease in Vietnam.pdf4. An Overview of Sugarcane White Leaf Disease in Vietnam.pdf
4. An Overview of Sugarcane White Leaf Disease in Vietnam.pdf
 
Deep Behavioral Phenotyping in Systems Neuroscience for Functional Atlasing a...
Deep Behavioral Phenotyping in Systems Neuroscience for Functional Atlasing a...Deep Behavioral Phenotyping in Systems Neuroscience for Functional Atlasing a...
Deep Behavioral Phenotyping in Systems Neuroscience for Functional Atlasing a...
 
insect taxonomy importance systematics and classification
insect taxonomy importance systematics and classificationinsect taxonomy importance systematics and classification
insect taxonomy importance systematics and classification
 
Multi-source connectivity as the driver of solar wind variability in the heli...
Multi-source connectivity as the driver of solar wind variability in the heli...Multi-source connectivity as the driver of solar wind variability in the heli...
Multi-source connectivity as the driver of solar wind variability in the heli...
 
RNA INTERFERENCE: UNRAVELING GENETIC SILENCING
RNA INTERFERENCE: UNRAVELING GENETIC SILENCINGRNA INTERFERENCE: UNRAVELING GENETIC SILENCING
RNA INTERFERENCE: UNRAVELING GENETIC SILENCING
 
Astronomy Update- Curiosity’s exploration of Mars _ Local Briefs _ leadertele...
Astronomy Update- Curiosity’s exploration of Mars _ Local Briefs _ leadertele...Astronomy Update- Curiosity’s exploration of Mars _ Local Briefs _ leadertele...
Astronomy Update- Curiosity’s exploration of Mars _ Local Briefs _ leadertele...
 

Centrifugation- Techniques in Biochemistry

  • 2.  A centrifuge is a device for separating particles from a solution according to their size, shape, density, viscosity of the medium and rotor speed.  Action of centrifugal force is used to promote accelerated settling of particles in a solid-liquid mixture.  Biological centrifugation is a process that uses centrifugal force to separate and purify mixtures of biological particles in a liquid medium. It is a key technique for isolating and analysing cells, subcellular fractions, supramolecular complexes and isolated macromolecules such as proteins or nucleicacid.
  • 3.  In a solution, particles whose density is higher than that of the solvent sink (sediment), and particles that are lighter than it float to the top. The greater the difference in density, the faster they move. If there is no difference in density (isopyknic conditions), the particles stay steady. To take advantage of even tiny differences in density to separate various particles in a solution, gravity can be replaced with the much more powerful “centrifugal force” provided by a centrifuge
  • 4. PRINCIPLE OF CENTRIFUGATION Basis of separation:  -Size  -Shape  -Density Methodology:  -Utilizes density difference between the particles/macromolecules and the medium in which these are dispersed  -Dispersed systems are subjected to artificially induced gravitational fields
  • 5.
  • 6. RELATIVE CENTRIFUGAL FORCE  the ratio of the centrifugal acceleration at a specified radius and the speed to the standard acceleration of gravity  RCF =42 (rev min -1 ) 2 r/ 3600 x981= G/g  RCF units are therefore dimensionless (denoting multiples of g) and revolutions per minute are usually abbreviated as r.p.m.: RCF=1.12 x 10-5 r.p.m2 r.
  • 8. GENERAL STEPS IN BIOCHEMICAL SEPARATION
  • 12. CLINICAL LABORATORY USE OF CENTRIFUGATION  Remove cellular components from blood to provide cell free plasma or serum for analysis.  Concentrate cellular elements and other components of biological fluid for microscopic or chemical analysis.  Remove chemically precipitated protein from an analytical specimen.  Separate protein bound or antibody bound ligand from free ligand in immunochemical assay.  Extract solute in biological fluids from aqueous to organic solvents.  Separate lipids compounds such as chylomicrons from other compounds in plasma or serum or lipoprotein from one another.
  • 13. Types of Centrifuge  Fixed Angle Rotor  Swinging Buckcet Rotor  Ultra Centrifugation  Axial Centrifugation
  • 14. FIXED ANGLE ROTOR SWINGING BUCKET ROTOR  Sediment particles have only short distance to travel before pelleting at the angle of 25 or 40 degree.  Shorter run time.  The most widely used rotor type.  This is used to separate nuclei, mitochondria, nucleosome and isopycnic separations. Isopycnic separation is when biological particle and surrounding medium attain the equal level at the zero sedimentation rate.  Longer distance of travel may allow better separation, such as in density gradient centrifugation.  The particles sediment uniformly along the horizontal tube.  Easier to withdraw supernatant without disturbing pellet.
  • 15. ULTRACENTRIFUGATION AXIAL CENTRIFUGATION  A very high speed centrifuge with fixed head rotors.  Most important application is in separating lipoproteins.  This is available both in analytical and preparative models.  Allows tubes to spun in vertical orientation which is used in traditional centrifugation.