Cell Biology Part II

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 Contents

 General idea of cancer cells, carcinomas, Sarcomas, Lumphomas,

Leukemia,

 Characteristics of Cancer cells,

 Carcinogenesis - Mutation and Viral theories of Carcinogenesis,

Environmental causes of cancer.

 Freezing and Thawing of cells, Radiations in Cell environment (UV

radiations, photodynamic sensitization).

 Principle and application of Electron microscopy

 Principle and application of Centrifugation techniques, TLC and Gel

electrophoresis

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 General idea of cancer cells

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 Types of Cancers

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 Characteristics of cancer cells

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 Carcinogenesis – The Development of Cancer

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 Theories of carcinogenesis

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 Theories of carcinogenesis….
Viral theory of carcinogenesis

VIRUS

Viruses
Viral DNA may be inserted into a host cell’s DNA
May switch on a proto-oncogene
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May
 Theories of carcinogenesis….

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 Theories of carcinogenesis….

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 Environmental causes of Cancer

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 Radiations in Cell environment (UV radiations, photodynamic sensitization).
 The potential biological effects and damages caused by radiation depend on the
conditions of the radiation exposure. It is determined by:

1. quality of radiation
2. quantity of radiation
3. received dose of radiation
4. exposure conditions (spatial distribution

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 Radiations in Cell environment (UV radiations, photodynamic sensitization).

 Photosensitization is a reaction to light that is mediated by a light-absorbing

molecule, which is not the ultimate target.

 Photosensitization can involve reactions within living cells or tissues, or they can
occur in pure chemical systems.

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 Cell biology techniques: Cryopreservation, centrifugation, Electron microscopy, TLC, Electrophoresis

 Since water is the major component of all living cells , cellular

metabolism stops when all water in the system is converted
to ice.

 Ice forms at different rates during the cooling process.

 During slow cooling, freezing occurs external to the cell

before intracellular ice begins to form.
 As ice forms, water is removed from the extracellular environment and an osmotic imbalance occurs
across the cell membrane leading to water migration out of the cell.

 The increase in solute concentration outside the cell, as well as intracellularly, can be detrimental to
cell survival.

 If too much water remains inside the cell, damage due to ice crystal formation and re-crystallization
during warming can occur.

 Warming from the frozen state should occur as rapidly as possible until complete thawing is
achieved.

 To achieve rapid warming: frozen tubes containing cells are placed in water bath at 37°C.

 Cells frozen in plastic tubes will take longer to thaw than that in glass ampoules, and sometimes
gentle agitation of the tube during warming will accelerate the thawing process.

 As soon as the contents of the tube have been thawed, remove the tube from the water bath. To
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 Cell biology techniques: Cryopreservation, centrifugation, Electron microscopy, TLC, Electrophoresis

What is a Centrifuge?
A centrifuge is a piece of laboratory equipment, driven by a motor, which spins
liquid samples at high speed.

Working principle:
Centrifuges work by the sedimentation principle, where the centripetal
acceleration is used to separate substances of greater and lesser density. The
rate of sedimentation is dependent upon the applied centrifugal field (G) being
directed radially outwards, this is dependent on angular velocities of rotor and
radial distance of the particles from the axis of rotation.

APPLICATION:

1. The separation of sediments in urine

2. Separation of protein free filtrate
3. Washing of RBCs by normal saline etc.
4. Separation of enzyme extracts.
5. Separation of cells, cell organelles etc.

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Regents Biology
 Cell biology techniques: Cryopreservation, centrifugation, Electron microscopy, TLC, Electrophoresis

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 Cell biology techniques:Electron microscopy
What are electron microscopes?
Scientific instruments/microscopes that use a focused beam of electrons to examine objects
on a very fine scale.

What is electron microscopy?

Electron microscopy is the science and technology of using an electron beam to form a
magnified image.

Principle of microscopy:

Electron guns generates a stream of electrons under high vacuum which is accelerated towards
the specimen in the form of a thin, focused, monochromatic beam using metal apertures and
magnetic lenses leading to irradiation of the sample by the beam. Interactions of electron beam
with the sample are detected and transformed into an image.

Application of electron microscopy

• Topography – Surface features of an object/cell/

cell organelle.
• Morphology – Size and shape of particles
making up object/cell /cell organelle.
• Composition – Relative amount of elements and
compounds making up the object/cell/cell
organelle, pigments etc..
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g arranged in the object/biomolecules etc..
 Cell biology techniques: Cryopreservation, centrifugation, Electron microscopy, TLC, Electrophoresis

Application of electron microscopy

• Topography – Surface features of an object/cell/ cell organelle.

• Morphology – Size and shape of particles making up object/cell /cell organelle.
• Composition – Relative amount of elements and compounds making up the object/cell/cell
organelle, pigments etc..
• Structure – Crystallography. How atoms are arranged in the object/biomolecules etc..

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 Cell biology techniques: Cryopreservation, centrifugation, Electron microscopy, TLC, Electrophoresis

What is a Thin Layer chromatography/TLC?

TLC is a type of chromatographic technique that is used to separate & identify
the components of mixture under study.

Working principle:
Thin layer chromatography (TLC) is a widely-employed laboratory technique
that involves a stationary phase of a thin layer of adsorbent like silica gel,
alumina, or cellulose on a flat, inert substrate. Different compounds in the
sample mixture travel different distances according to how strongly they
interact with the adsorbent.

APPLICATION:

1. Separation of macromolecules
2. Separation of pigments .
3. Separation of fatty acids, triglycerides etc.

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 Cell biology techniques: Cryopreservation, centrifugation, Electron microscopy, TLC, Electrophoresis

What is a electrophoresis?

Electrophoresis is an analytical method frequently used in cell biology to

separate samples containing proteins , nucleic acids etc.

Working principle:
Charged molecules migrate in electric field to the electrodes of opposite
charge.

APPLICATION:

1. Separation and purification of macromolecules

2. Separation of proteins from serum, blood, urine, CSF .
3. Separation of nucleic acids.

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 Summary

1. Cancer cell properties, Cancer types depending on cells from which

tumors develop, Behavior of cancer cells over normal cells, Step-wise
Mechanism of cancer development, Mutation and Viral theories of
carcinogenesis, Environmental carcinogens

2. Effects of UV and photodynamic sensitization in the cell environment.

3. Techniques used in cell biology to explore and to study its components

such as Cryopreservation, Centrifugation, electron microscopy, thin layer
chromatography and electrophoresis were studied for its principle &
applications.

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 References
S.C. Rastogi (2003) Cell and Molecular Biology –New Age International Publishers, New Delhi.

M. P. Arora (2000) Molecular Biology’Himalaya Publishing House Pvt.Ltd. Mumbai.

A. C. Giese,(2003) Cell Physiology, 5th Edition, W.B. Saunders Company, Philadelphia PA, 1979.

C. B. Pawar (2004), cell Biology, Himalaya Publishing House, Mumbai

De Robertis and De Robertis, (2000) Cell and Molecular Biology, Lippincott Williams and Wilkins publishers

Verma and Agarwal (2004), Cell Biology, Genetic s, Molecular Biology, Evolution and Ecology, S. Chand publication,

New Delhi.

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http://www.ncbi.nlm.nih.gov/books/NBK26902/
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http://www.careflash.com/video/cancer-overview?lc=en
 http://www.ncbi.nlm.nih.gov/books/NBK9552/
http://www.ncbi.nlm.nih.gov/books/NBK9963/
http://www.ncbi.nlm.nih.gov/books/NBK9878/
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http://www.ncbi.nlm.nih.gov/books/NBK26902/
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video_5390a86c3.html#axzz3B6C9jyFj
http://www.learner.org/courses/biology/textbook/cancer/
http://www.learner.org/courses/biology/textbook/cancer/cancer_8.html
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http://www.cancerquest.org/cancer-biology-animations.html#
 THANK YOU

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