University Mohamed El Bachir El Ibrahimi of Bordj Bou Arreridj
Institute of Life & Natural Sciences & of Earth & Universe Sciences
Module of :
CELL BIOLOGY
« Lectures for First-Year Students »
Chapter II: Basic tools of cell biology
Prepared by:
Dr. CHENOUF N. S.
Academic Year: 2023/2024
Introduction
• Histology ( histo= tissues, logy = science) depends on the
laboratory methods that can be used to study cell structure
and function.
• In animals there are four tissue types:
• Epithelium tissue: is essentially composed of cells
covering all the surfaces of the body exposed to
the outside world and lining the outside of organs.
The largest example of epithelial tissue is the skin.
• Muscle tissue.
• Nerve tissue is made up of nerve cells (neurons).
• Connective tissue supports other tissues and binds
them together (bone, blood, and lymph tissues).
Tools of histology
• Some of the important basic tools of cell
biology are described in this chapter,
including: Microscopy (light & electron
microscopes), subcellular Fractionation
through centrifugation, cytochemical
methods and cell culture.
1- Microscopy
• The very discovery of cells arose from the development of
the microscope
• The English scientist Robert Hooke (1635 -1703) first coined
the term “cell” following his observations of a piece of cork
with a simple light microscope in 1665
1- Microscopy
• Using a microscope that magnified objects up to
about 300 times their actual size, the Dutch
scientist Antony van Leeuwenhoek, in the 1670s,
was able to observe a variety of different types of
cells, including sperm, red blood cells, and
bacteria.
1-1 Light Microscopy (LM)
• LM remains a basic tool of cell biology
LM uses light as illuminating
source
Magnification
system (up to 1500X)
We use colored stains to
selectively highlight particular
structures and organelles. The
image is colored
1-1 Light Microscopy (LM)
• Magnification power = the degree of enlargement
• Resolution power: the ability of a microscope to
distinguish objects separated by small distances
(Low resolution: 0.2 μm)
• With LM, the cell and some of the larger
subcellular organelles, such as nuclei,
chloroplasts, and mitochondria can be
observed.
Summary of Visible Cell Features under Light
Microscopy
Some interesting stains
• H&E staining is the staining of tissue sections (will be
explained in the TD session) with Haematoxylin, which
stains cell nuclei blue, and Eosin, which stains the
extracellular matrix, cytoplasm and other structures varying
shades of pink. This allows visualisation of the structure,
distribution of cells and morphological changes within a
tissue sample.
Some interesting stains
- Janus Green B is used to stain mitochondria
- Silver stain is used to visualize the Golgi
system and nervous cells
1-2 Electron Microscopy (EM)
• The electron microscope can achieve a much
greater resolution than that obtained with
the light microscope (High 0,2 nm)
• 1 mm= 1000 μm
• 1 μm = 1000 nm
• 1 nm = 10 angstroms (Å)
• Magnification power : up to 1000 000 X
1-2 Electron Microscopy (EM)
• EM uses electrons as illuminating source
• The use of osmium tetroxide fixation to
preserve lipids is fundamental to EM
Electron dense
Electron lucent
LM VS EM
Cell structure as seen through light and transmission electron microscopes
1-2 Electron Microscopy (EM)
• There are two main types of EM:
Transmission EM (TEM) is used to view thin specimens (tissue
sections, molecules, etc) through which electrons can pass
generating a projection image.
Scanning EM (SEM) provides detailed images of the surfaces of
cells and whole organisms that are not possible by TEM. It can
also be used for particle counting and size determination, and for
process control.
2- Subcellular Fractionation
To address many of the questions concerning the
function of subcellular organelles, it has proven
necessary to isolate the organelles of eukaryotic cells
in a form that can be used for biochemical studies.
This is usually accomplished by differential
centrifugation, to separate the components of cells
on the basis of their size and density.
2- Subcellular Fractionation
3- Cytochemical methods
• Cytochemistry is the science of localizing
chemical components of cells and organelles
(sugars, fatty acids, amino acids, and
nucleotides) on histological sections (a
histological section refers to a thin slice of
tissue that is viewed under a microscope), by
using various techniques.
3- Cytochemical methods
3-1 Chemical methods
• Staining of DNA with Feulgen reaction (will
be explained in DW session),
• Staining of proteins with Millon reactions
• Staining of glucides with the periodic acid-
Schiff (PAS) reaction, … etc.
3- Cytochemical methods
4-2 Physical methods
- The changes of temperature of specimens
such as microincineration at high
temperatures and cryo-techniques at low
temperatures,
- Effects of wavelength on absorption such as
microspectrophotometry,
- Utilization of radiation such as
radioautography and X-ray microanalysis.
4- Cell culture
• Cell culture refers to laboratory methods that
enable the growth of eukaryotic or prokaryotic
cells in physiological conditions.
• It is most commonly linked to creating model
systems that study basic cell biology, replicate
disease mechanisms, or investigate the toxicity of
novel drug compounds.
4- Cell culture
• Animal cell cultures are widely used to
prevent animal testing for toxicity testing of
new drugs, chemicals, and cosmetics. Major
animal cell cultures used in this domain are
Kidney- and liver-derived cell cultures.
4- Cell culture
• Microbial cell culture
Nota bene: There are many other techniques used in cell biology.