Biochemical axis. Centrifuge Machine Types of centrifugation

Biochemical
techniques

v Spectrophotometer

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v Chromatography

v ELISA

v Centrifugation

v Radioactivity

v Electrophoresis

v Microscopy

 

Spectrophotometer

Spectrophotometer is a method to
measure a chemical absorbs light by measuring the intensity of light as a beam
of light passes through solution.

          OR

Spectrophotometer is the quantitative
measurement of the reflection or transmission properties of a material as a
function of wave length .

Principle of  Spectrophotometer

 The principle of spectrophotometer is based on
lambert’s, beer’s law.

Spectrophotometer

Centrifuge

Is a device for separating particles from a
solution according to there size, shape, viscosity, density of the medium.

Principle of centrifugation

An object moving in a circle at a steady
angular velocity will experience a force F .this is the basis of centrifugation
.contain velocity and the radius of rotation collectively determine the
magnitude of the force.

Centrifugation is performed in a centrifuge
.a centrifuge is a mechanical device produces a centrifugal force by spinning
around a central axis.

                           

Centrifuge Machine

 

Types of centrifugation

  Bench
centrifuge .

  Continous
flow centrifuge.

  High
speed refrigerated centrifuge.

  Ultra
centrifuge.

 

Some types of centrifugation are
defined here,

·      
Bench
centrifugation

Commonest type of centrifugation .

·      
Ultra
centrifugation

High speed spinning that separate
out colloidal and small particles in sample to separate components of sample
based on shape and mass

ELISA

Is
based on affinity.

An ELISA test can be  detect both current and past infections.as
well as antibodies ,an ELISA test can also be used to detect antigens or
enzymes.

Principle of
ELISA

 the technique involves three
principles:

·        
An
immune reaction i.e.
antigen-antibody reaction.

·        
Enzymatic
chemical reaction i.e.
formation of colored (chromogenic) product from colorless substrate.

·        
Signal
detection and Quantification i.e.
detection of color intensity of the colored products by the enzyme and add
substrate.

 

ELISA –An enzyme –linked immune assay (ELISA)
is a test that detects antibodies in the blood.

ELISA Machine

 

Types of ELISA

  Indirect
ELISA.

  Direct  ELISA.

  Competitive
ELISA

  Sandwich
ELISA.

Applications

o  ELISA  test is used in food industries to detect food
allergens.

o  ELISA
is used to detect antigen and antibody in blood sample to diagnose for past
exposure of disease.

o  ELISA
Is used in home pregnancy test.

o  ELISA
is also used in monoclonal antibody screening.

o  This
technique is also used in virus testing.

 

Electrophoresis

The movement of charged particle an electric
field resulting their migration towards the oppositely charged electrode .

Or

Apply
electric current to separate molecule on the basis of their charge.
Electrophoresis perform using different types of support media .

 

Electrophoresis  Types

  Paper
Electrophoresis .

  Gel
Electrophoresis .

  Immune
Electrophoresis .

  Isoelectric
Focusing .

  Capillary
Electrophoresis.

 

Some types of electrophoresis is
defined here,

Gel
electrophoresis

Is used to separate larger
molecules of DNA .

Capillary
electrophoresis

It is combined the high
resolution of electrophoresis with the speed of high performance liquid
chromatography .and it is used to separate amino acids ,protein, and DNA.

 Iso
electric focusing

It is used to separate molecules
on the basis of differences in their electric point.

 

Applications

o  Electrophoretic
techniques have many applications in biomedical sciences such as investigation
of diseases and multiple sclerosis.etc

 

 

Chromatography

Is used to separate closely  related compound from a mixture. It is no
longer limited to the separation of plant pigment s such as chlorophyll.  

         Or

Chromatography is a physical
method of separation of the component s of a mixture.

Chromatography is useful for a
wide range of separation processes and chemical analysis tasks especially in
biochemistry.

Chromatography Principle

Is based on the principle of the
partition of the solute between two phases.

 

 

these two phases are :


Mobile Phase

Mixture of the substances to be
separated dissolved in a liquid or a gas.


Stationary Phase

Is a porous solid matrix.

The interaction between the
mobile and the stationary phases  result
in the separation of the compound from the mixture.

Chromatograph

Chromatography  Types

  Paper Chromatography
.

  Thin Layer
Chromatography.

  Column Chromatography.

  Exchange Chromatography.

  Affinity Chromatography.

  HPLC.

 

Paper
chromatography

Simplest  type of chromatography .it does not always
give accurate results.

Thin layer
chromatography

In this technique we used thin
layer of cellulose or silica coated on to a plastic or glass sheet.it gives
much better and more reliable results.

Column
chromatography

In this technique we used glass
column filled with a cellulose slurry .

High
performance liquid chromatography

HPLC is a improved and defined
form of column chromatography.

Applications


Chromatographic technique is useful in
identifying components of mixture and also provide quantitative data.


Chromatography is used to investigate
criminal cases and help to catch criminals.


Recently,scientist is used Chromatography
to  determining various antibodies to
fight against Ebola virus.


This technique is also used in food testing,
beverage testing and also in drug testing.In drug testing this technique
quantify a drug and their metabolites in blood and urine.


Gas 
and liquid chromatography together trace analysis of organic and
inorganic compounds in a sample.

 

Radioactivity

Spontaneous emission of particles
,rays,or waves of energy from the unstable nuclei of some atoms.these atoms are
known as Radioisotopes.and when they decay they form more stable atoms. The
rays, waves and particles are called radiation.

 Radiation is the energy released when an
unstaible atom change to a stable form.

      Or

Radiation is the emission of
energy as electromagnetic wave these waves are called photons.

There are three different types
of radiations.

·       Alpha
.

·       Beta.

·       Gamma
.

These radiations released by unstable nuclei untile they reach a
stable state.all of these radiations are detect and measure easily.

 

 

 

 

 

 Detection Of Radioactivity

Radioactivity
is detect and measure by excitation of electrons. alpha particle raditions are
used in biomedical science. to detect beta and gamma rays three methods are
available which is

o  Gas Detector
.

o  Liquid
Scintillation Counting.

o  Autoradiography.

 

 

Applications


Radiation is harmful .precautions must be
observed when using radioisotopes.


It is used in biological and medical research
.Biochemical
techniques

v Spectrophotometer

v Chromatography

v ELISA

v Centrifugation

v Radioactivity

v Electrophoresis

v Microscopy

 

Spectrophotometer

Spectrophotometer is a method to
measure a chemical absorbs light by measuring the intensity of light as a beam
of light passes through solution.

          OR

Spectrophotometer is the quantitative
measurement of the reflection or transmission properties of a material as a
function of wave length .

Principle of  Spectrophotometer

 The principle of spectrophotometer is based on
lambert’s, beer’s law.

Spectrophotometer

Centrifuge

Is a device for separating particles from a
solution according to there size, shape, viscosity, density of the medium.

Principle of centrifugation

An object moving in a circle at a steady
angular velocity will experience a force F .this is the basis of centrifugation
.contain velocity and the radius of rotation collectively determine the
magnitude of the force.

Centrifugation is performed in a centrifuge
.a centrifuge is a mechanical device produces a centrifugal force by spinning
around a central axis.

                           

Centrifuge Machine

 

Types of centrifugation

  Bench
centrifuge .

  Continous
flow centrifuge.

  High
speed refrigerated centrifuge.

  Ultra
centrifuge.

 

Some types of centrifugation are
defined here,

·      
Bench
centrifugation

Commonest type of centrifugation .

·      
Ultra
centrifugation

High speed spinning that separate
out colloidal and small particles in sample to separate components of sample
based on shape and mass

ELISA

Is
based on affinity.

An ELISA test can be  detect both current and past infections.as
well as antibodies ,an ELISA test can also be used to detect antigens or
enzymes.

Principle of
ELISA

 the technique involves three
principles:

·        
An
immune reaction i.e.
antigen-antibody reaction.

·        
Enzymatic
chemical reaction i.e.
formation of colored (chromogenic) product from colorless substrate.

·        
Signal
detection and Quantification i.e.
detection of color intensity of the colored products by the enzyme and add
substrate.

 

ELISA –An enzyme –linked immune assay (ELISA)
is a test that detects antibodies in the blood.

ELISA Machine

 

Types of ELISA

  Indirect
ELISA.

  Direct  ELISA.

  Competitive
ELISA

  Sandwich
ELISA.

Applications

o  ELISA  test is used in food industries to detect food
allergens.

o  ELISA
is used to detect antigen and antibody in blood sample to diagnose for past
exposure of disease.

o  ELISA
Is used in home pregnancy test.

o  ELISA
is also used in monoclonal antibody screening.

o  This
technique is also used in virus testing.

 

Electrophoresis

The movement of charged particle an electric
field resulting their migration towards the oppositely charged electrode .

Or

Apply
electric current to separate molecule on the basis of their charge.
Electrophoresis perform using different types of support media .

 

Electrophoresis  Types

  Paper
Electrophoresis .

  Gel
Electrophoresis .

  Immune
Electrophoresis .

  Isoelectric
Focusing .

  Capillary
Electrophoresis.

 

Some types of electrophoresis is
defined here,

Gel
electrophoresis

Is used to separate larger
molecules of DNA .

Capillary
electrophoresis

It is combined the high
resolution of electrophoresis with the speed of high performance liquid
chromatography .and it is used to separate amino acids ,protein, and DNA.

 Iso
electric focusing

It is used to separate molecules
on the basis of differences in their electric point.

 

Applications

o  Electrophoretic
techniques have many applications in biomedical sciences such as investigation
of diseases and multiple sclerosis.etc

 

 

Chromatography

Is used to separate closely  related compound from a mixture. It is no
longer limited to the separation of plant pigment s such as chlorophyll.  

         Or

Chromatography is a physical
method of separation of the component s of a mixture.

Chromatography is useful for a
wide range of separation processes and chemical analysis tasks especially in
biochemistry.

Chromatography Principle

Is based on the principle of the
partition of the solute between two phases.

 

 

these two phases are :


Mobile Phase

Mixture of the substances to be
separated dissolved in a liquid or a gas.


Stationary Phase

Is a porous solid matrix.

The interaction between the
mobile and the stationary phases  result
in the separation of the compound from the mixture.

Chromatograph

Chromatography  Types

  Paper Chromatography
.

  Thin Layer
Chromatography.

  Column Chromatography.

  Exchange Chromatography.

  Affinity Chromatography.

  HPLC.

 

Paper
chromatography

Simplest  type of chromatography .it does not always
give accurate results.

Thin layer
chromatography

In this technique we used thin
layer of cellulose or silica coated on to a plastic or glass sheet.it gives
much better and more reliable results.

Column
chromatography

In this technique we used glass
column filled with a cellulose slurry .

High
performance liquid chromatography

HPLC is a improved and defined
form of column chromatography.

Applications


Chromatographic technique is useful in
identifying components of mixture and also provide quantitative data.


Chromatography is used to investigate
criminal cases and help to catch criminals.


Recently,scientist is used Chromatography
to  determining various antibodies to
fight against Ebola virus.


This technique is also used in food testing,
beverage testing and also in drug testing.In drug testing this technique
quantify a drug and their metabolites in blood and urine.


Gas 
and liquid chromatography together trace analysis of organic and
inorganic compounds in a sample.

 

Radioactivity

Spontaneous emission of particles
,rays,or waves of energy from the unstable nuclei of some atoms.these atoms are
known as Radioisotopes.and when they decay they form more stable atoms. The
rays, waves and particles are called radiation.

 Radiation is the energy released when an
unstaible atom change to a stable form.

      Or

Radiation is the emission of
energy as electromagnetic wave these waves are called photons.

There are three different types
of radiations.

·       Alpha
.

·       Beta.

·       Gamma
.

These radiations released by unstable nuclei untile they reach a
stable state.all of these radiations are detect and measure easily.

 

 

 

 

 

 Detection Of Radioactivity

Radioactivity
is detect and measure by excitation of electrons. alpha particle raditions are
used in biomedical science. to detect beta and gamma rays three methods are
available which is

o  Gas Detector
.

o  Liquid
Scintillation Counting.

o  Autoradiography.

 

 

Applications


Radiation is harmful .precautions must be
observed when using radioisotopes.


It is used in biological and medical research
.


It is also used in diagnostic purpose .and
treat some types of clinical conditions.

 

 


It is also used in diagnostic purpose .and
treat some types of clinical conditions.

 

 

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