Cell structure

(Nucleus + Cytoplasm)

Dr.Mohammad Omran, MD
Arab American University
Faculty of medicine
Polarizing Microscopy
● Polarizing microscopy allows the recognition of stained or unstained
structures made of highly organized subunits.

● When normal light passes through a polarizing filter, it exits vibrating in only
one direction. If a second filter is placed in the microscope above the first
one, with its main axis perpendicular to the first filter, no light passes through.

● If, however, tissue structures containing oriented macromolecules are located

between the two polarizing filters, their repetitive structure rotates the axis of
the light emerging from the polarizer and they appear as bright structures
against a dark background
birefringence

● Is the ability to rotate the direction of vibration of polarized light

● A feature of crystalline substances or substances containing highly
oriented molecules, such as cellulose, collagen, microtubules, and
actin filaments
AUTORADIOGRAPHY
● A method of localizing newly synthesized macromolecules in cells or tissue
sections
Steps:
•Label metabolites (nucleotides, amino acids, sugars) with radioactive substances
•When specific macromolecules (DNA, RNA, protein, glycoproteins, and
polysaccharides) are synthesized in the cell, the labeled metabolite will be
incorporated.
•Tissue fixation, processing, and sectioning
•only the new, labeled macromolecules continue to emit weak and localized
radiation
•Slides or TEM grids with radiolabeled cells or tissue sections are coated in a
darkroom with photographic emulsion in which silver bromide crystals act as micro
detectors of the radiation.
•After an adequate exposure time in light proof boxes, the slides are developed
photographically.
• Silver bromide crystals reduced by the radiation produce small black grains of
metallic silver, which under either the light microscope or TEM indicate the
locations of radiolabeled macromolecules in the tissue.
Importance:
● Know which cells are replicating
● where in the cell protein is produced, if it is secreted, and its path in the
cell before being secreted?
Autoradiographs from the salivary gland of a mouse injected with 3H-fucose 8
hours before tissue fixation.
Fucose was incorporated into oligosaccharides, and the free 3H-fucose was
removed during fixation and sectioning of the gland.
reveal locations of newly synthesized glycoproteins containing that sugar.
(a) Black grains of silver from the light-sensitive material coating the specimen are
visible over cell regions with secretory granules and the duct indicating
glycoprotein locations. (X1500)
(b) The same tissue prepared for TEM autoradiography shows silver grains with a
coiled or amorphous appearance again localized mainly over the granules (G) and
in the gland lumen (L). (X7500)
Cell and tissue culture
•In vivo: in the living
organism
•In vitro: outside the body of
a living organism
• tissue or organ and placed with sterile
Cell culture:
Live cells and tissues can be
maintained and studied
outside the body in culture (in
vitro by bathing the cells in
fluid derived from blood
plasma and containing many
different molecules required
for survival and growth.
Primary cell culture:
Cells to be cultured are dispersed
mechanically or enzymatically from a
procedures in a clear dish to which they
adhere, usually as a single layer.
Cell lines:
Some cells can be maintained in vitro for
long periods because they become
immortalized and constitute a permanent
cell line
Medical application of cell and tissue cultures
Main principle of IHC

https://www.youtube.com/watch?v=bfeNUmqHxyc
Medical applications of enzyme histochemistry
Visualizing Specific Molecules
Visualizing Specific Molecules
The most commonly used labels:
•Fluorescent compounds
•Radioactive atoms that can be detected with autoradiography
•Molecules of peroxidase or other enzymes that can be detected with
histochemistry
•Metal (usually gold) particles that can be seen with light and electron
microscopy.
 Cell structure
(Nucleus + Cytoplasm)
 Cell structure
•Outlines:
1.Cell membrane structure and function
2.Cell signaling and transduction
3.Cell organelles
4.Karyotyping and Barr bodies
5.Cell cycle checkpoints
6.Meiosis and mitosis
7.Cell death
8.Stem cell and renewal
Cell membrane
•Structure?
•Function of each macromolecule ?
•Whole function of cell membrane as unit
membrane?
 Cell Structure and function

https://www.youtube.com/watch?v=URUJD5NEXC8
Structure and Function of Cell (Plasma) Membrane
⚫ Structure : 7.5 to 10 nm in thickness
◦ Membrane lipids
◦ Membrane proteins
◦ Membrane carbohydrates
- attached to proteins or lipids
- glycoproteins or glycolipids
🡺 important components of RECEPTORS
⚫ Function
◦ Physical isolation
◦ Regulation of exchange with environment
◦ Sensitivity
◦ Structural support
◦ Communication
◦ Electrochemical gradient : determines the direction that ions will flow
through an open ion channel
Types of proteins in the cell membrane
● Integral Proteins
- incorporated or embedded in the lipid layers
- when completely span the lipid bilayer 🡺 they
are called transmembrane proteins
● Peripheral Proteins
- loosely bound to the surface of membrane
Defective receptors in cell membrane
Cell signaling and transduction
Cytoplasmic organelles
•10 hours old male patient
presented with yellowish
discoloration of skin and sclera.
•Maternal blood group: O+ve.
•Baby’s blood group: A+ve
•What is the organelle that is
responsible for metabolism of
bilirubin?
ER
 Golgi Apparatus
Functions
1. Modifying, sorting, and packaging cell
macromolecules for cell secretion (exocytosis) or for
use within the cell.
2. It modifies (compresses) proteins delivered from the
rough ER
3. involved in the transport of lipids around cell.
4. It forms lysosomes.
5. Synthesis of proteoglycans molecules present in the
extracellular matrix and carbohydrates.
Lysosomes
⚫ Are organelles formed by Golgi that contain digestive
enzymes (acid hydrolases).
⚫ The membrane surrounding a lysosome prevents the digestive
enzymes inside from destroying the cell. Lysosomes fuse with
vacuoles and dispense their enzymes into the vacuoles, digesting
their contents.
Lysosome
Defective lysosomal function
Proteasomes
•Function:
1.Degradation of denatured or
nonfunctional polypeptides.
2.Removal of proteins no
longer needed by the cell.
3. providing an important
mechanism for restricting the
activity of a specific protein to a
certain window of time .
Mitochondria
Peroxisome
⚫ Similar to lysosomes
⚫ Large vesicles containing oxidative enzymes which
transfer H from substrates to O
⚫ Contains catalase that changes H2O2 to H2O
 The signs and symptoms of
Zellweger syndrome typically
appear during the newborn
period and may include :
1. poor muscle tone
(hypotonia)
2. poor feeding
3. Seizures
4. hearing loss, vision loss
Abnormal clinical features in neonatal
5. distinctive facial features
adrenoleukodystrophy are generally
skeletal abnormalities.
present at birth, and include:
1. muscle hypotonia
2. severe psychomotor retardation
3. failure to thrive
Cytoskeleton
Cell Inclusions
cytoplasmic structures or deposits filled with stored
macromolecules and are not present in all cells.
Nucleus
Nucleus
 Barr bodies
• 35 year old male presented with his wife to fertility clinic with history
of primary infertility.
• On examination: he is tall , bilateral gynecomastia , hypogonadism
• What do you think is the diagnosis?
• How do we confirm it?
hermaphroditism, the condition of having both male and female
reproductive organs.

A pseudohermaphrodite is a person whose gonads are consistent

with the chromosomal sex but who has external genitalia of the
opposite sex.
 Karyotyping
38 year old female , Gestational age
14+2 wks
There is first degree consanguinity
between the 2 parents.
patient with history of giving birth to a
dysmorphic 2 neonates both died at age
of 1 month .
• What do you think the cause of this?
• What do you think best next step of
management?
 Karyotyping

• Karyotypes provide light microscopic information regarding the number and morphology
of chromosomes in an organism.
• Such preparations are made by staining and photographing the chromosomes of a
cultured cell arrested during mitosis, when chromosomes are maximally condensed.

• From the image individual chromosomes are typically placed together in pairs. With
certain stains, each chromosome has a particular pattern of banding that facilitates its
identification and shows the relationship of the banding pattern to genetic anomalies.
Hybridization with fluorescent probes specific for each chromosome (FISH) followed by
karyotyping yields an image like that shown above.
• Note that the 22 pairs of autosomes, as well as the X and Y chromosomes, differ in size,
morphology, and location of the centromere.
 Check points
• 4 months old male patient
presented to clinic for
evaluation.
• O/E he has upwards slanting of
eyes , sandal sign , single simian
crease , systolic murmur
• What do you think his genetic
abnormality is?
• Diagnosis?
 Check points

1. Start G1/S checkpoint.

2. Unreplicated DNA checkpoint.
(G2/M)
3. Metaphase/anaphase
checkpoint
Mitosis
The Cell Cycle

https://www.youtube.com/watch?v=e6N9_RhD10Q
M Phase of the Cell Cycle

https://www.youtube.com/watch?v=5bq1To_RKEo
 Cell death

• Apoptosis :
• It is the process of cell suicide .
• is a rapid, highly regulated cellular
activity that shrinks and eliminates
defective and unneeded cells.
• results in small membrane-enclosed
apoptotic bodies, which quickly
undergo phagocytosis
Stem cells and cell renewal