• Recognise the staining characteristic of some of the more commonly used histochemical stains

Acidophilic - attracts acid dyes such as Haematoxylin (stains blue)

Basophilic - attracts basic dyes such as Eosin (stains pink)

Haematoxylin and Eosin (H+E) - Nuclear chromatin - blue to purple, Cytoplasm, collagen, keratin,
erythrocytes – pink, Cartilage - bluish

Aldehyde-fuchsin - Elastic fibres, mast cell granules, mucus – deep purple

Van Geison’s picro fuchsin technique - Collagen – pink to red, Muscle, erythrocytes – yellow,
Nuclei – blackish

Masson’s Trichrome - Nuclei – blue/back, Collagen, mucin – blue or green, Cytoplasm - pink,
muscle, keratin – red

Silver stain for neural tissue and bone - ground section

Von kossa’s silver method - Calcium, basement membranes –brown to black, Nuclei – red

Periodic acid Schiff (PAS) - Cartilage, glycogen, mucin, basement membranes – purple

Sudan Black
Lipids – black

Cason’s Trichrome
Nuclei, cytoplasm – red, Erythrocytes – orange, Collagen – blue

• Perform simple linear measurements and calculations on cells and structures

Microscopy measurements are expressed in micrometres (μ) or nanometres (nm).

1mm = 1000μm
1μm = 1000nm

How to calculate the magnification of a micrograph

Use a ruler to measure the length of the scale bar in millimetres and convert this to μ. Then using
the equation for calculating magnification...
!"#$ &' "()*$
Magnification = )+,-). !"#$ &' &/0$+,
How to calculate to size of a histological structure
Use a ruler to measure the diameter, in millimetres, of an erythrocyte (R) in the micrograph.
Convert this length to micrometres.
From the previous calculation we already know the magnification of the micrograph. Rearrange
the magnification equation to find the actual diameter of the erythrocyte.

How to draw a scale bar on your image

The above image was taken at a magnification of 1,500x

The known diameter of an erythrocyte is between 6-8μ (7μ average)
You want to draw a scale bar representing an actual length of 10μ. How long should the drawing
of your scale bar be?
• Mentally reconstruct a 3D structure from 2D images

• Describe in detail the structure of the membranous and non-membranous organelles and relate
their structure to function

Organelle Structure Function Memb./Non-

memb.
Ribosomes • Round shaped • Synthesizing proteins in Non-
organelles the cell membranous
Microtubules • Tubulin protein • Intracellular support Non-
• Hollow cylinders • Cell shape membranous
• 22nm diameter • Cilia movement
• 5nm thick walls • Chromosome
• dynamic arrangement
Microfilaments • Actin • Microvilli structure Non-
• Helical array • Extension of cell membranous
• 6nm diameter processes
• Flexible
Intermediate • 8nm diameter • Provide mechanical Non-
Filaments • Heterogeneous strength & resistance to membranous
extracellular forces
Centrosome • Consists of 2 tiny • Play an important role in Non-
particles called the formation of flagella membranous
centrioles & cilia & cell division
(centrosome)
• Each centriole
consists of 9 groups
of microtubules
Rough ER • Presence of large no. • Protein synthesis via Membranous
of ribosomes on its transcription &
surface translation
• Cisternae, • Making changes on
ribosomes, nucleus, proteins produced by
nissl bodies ribosomes
 • Making new membranes
in the cell
Smooth ER • Presence of a fewer • Lipid & glycogen Membranous
no. of ribosomes metabolism
• Cisternae + glycogen • Synthesize & secrete
steroids
• Isolates Ca++
• Detoxification
• Membrane formation
Golgi Apparatus • Crescent shape of • Post-translational Membranous
stacked, flattened modification, sorting &
membrane bound packaging of proteins
cisternae with small • Distributes these
vesicles budding off substances within the
the cisternae cell to organelles called
lysosomes, to the
plasma membrane of
the cell and to outside
of the cell as secretory
products
Mitochondria • Sac-like • Generate ATP via Membranous
• Outer membrane --> oxidative
• Intermembrane phosphorylation
space --> critric acid cycle
• Inner membrane --> beta-oxidation of
forms cristae and fatty acids
elementary particles • Initiate apoptosis via
• Matrix - enzymes + release of cytochrome c
Ca++ into the cell cytoplasm
Endosomes • Membrane-bound • Involved in Membranous
• Formed during internalization of
endocytosis and via extracellular material &
the golgi apparatus recycling of membranes
Lysosomes • Membrane-bound • Contains hydrolytic Membranous
• Formed from golgi & enzymes and are
endsomes involved in intracellular
digestion (proteases,
nucleases, lipases…)
• Common in
macrophages &
neutrophils

*The number of Golgi bodies in the cell differ according to the cell's secretion activity.
*Endosomes & Lysosomes are involved in the internalization of extracellular material, recycling of
membranes & breakdown of macromolecule.

• Draw and label schematic diagrams of these organelles

• Describe the components of the eukaryotic cell nucleus

Structure of the NUCLEUS

• Nuclear Chromatin stains differently because it is packed more or less tightly.
• Each chromosome in the interphase nucleus is considered to consist of a single chromatin
filament which is tightly coiled and of considerable length.
• Chromatin consists of about 40% DNA, 40% protein called histone and 20% globular and acid
proteins with a little RNA.
• A filament of chromatin consists of a very long DNA molecule.
• The filaments have a diameter of 11nm and consist of subunits called Nucleosomes.
• Nucleosomes have a central region consisting of 2 molecules from each of the 4 histone
proteins. The DNA helix (140 base pairs) then wraps around this central histone bundle.
• Adjacent nucleosomes are held together by links of DNA (50 base pairs long) plus one
histone protein.
• The DNA in the links is continuous with the DNA of the nucleosomes.
• The function of the nucleosome is to reduce the length of DNA in a chromosome.
• The nucleosome represents a reduction of about 10-fold over a fully extended DAN helix.
• Nucleosomes are then folded into fibres of 30nm in diameter, called the solenoid.
• The solenoid represents a 40-fold reduction in length.
• In the metaphase nucleus, the DNA is packed approximately 10,000x more tightly than in the
extended DNA helix.

• Differentiate between euchromatin and heterochromatin

Euchromatin Heterochromatin
More dispersed chromatin Highly condensed chromatin
Active inactive
Pale areas Dark areas
Not evident in the light Stains readily with haematoxylin
microscope and other basic dyes
Present within the 'clear' areas
between the heterochromatin
• Describe the ultrastructure of the plasma membrane and relate structure to function
• Lipid-bilayer structure
• 9nm wide
• Surrounds the cell

• Selectively permeable
• Dynamic, fluid boundary
• Lipids & proteins
• Pumps, channels, receptors…

• Classify epithelia based on the number of cell layers and the shape of the surface cells and relate
to function

Types of Epi. Cell Shape & Layers Function Location

Tissue
Simple • A single layer of • Absorption, secretion
epithelium cells & filtration facilitated
by the single layer of
cells
Simple squamous • Flat in shape • Single layer thin • Found in the walls
• Arranged in single enough to form a of capillaries,
layer membrane for passive linings of the
diffusion pericardium &
linings of the alveoli
of the lungs
 Simple cuboidal • Single layer cells • Secretion & • Small collecting
• Tall as they are absorption ducts of the
wide kidneys, pancreas &
salivary glands
Simple Columnar • Single row of tall, • High secretory • Wall of the
closely packed function stomach
cells, aligned in a • Absorptive areas • Small intestine
row
• Cellular extensions
Pseudostratified • nuclei appear at • Ciliated epi. found
different heights in the airways
• fine hair-like (nose, bronchi)
extensions of their • also found in the
apical membrane - uterus and fallopian
cilia tubes of females -
• Columnar epi. Cell cilia propel ovum to
+ cuboidal epi. Cell the uterus
+ basal epi. Stem
cell
Stratified • Multilayered • Durable • Found where body
Epithelium • Can be columnar, • Protection linings have to
cuboidal or withstand
squamous mechanical of
chemical insults
Transitional • Stratified • Found in tissues
Epithelia • Cuboidal, that stretch
columnar & dome- • Exclusively found in
shaped cells the bladder, ureters
and urethra

• Understand the concept of cell polarity

• Cell polarity refers to the intrinsic asymmetry observed in cells, either in their shape, structure, or
organization of cellular components
• Epithelial cells establish an apical-basal polarity

Apical
• Directed towards lumen
• Surface modifications

Lateral
• Communication
• Attachment cell to cell
• Barrier

Basal
• Communication
• Attachment to connective tissue
• Identify and describe the structural surface modifications on the apical plasma membrane and
relate their structure to function

• Identify and describe the modifications on the lateral plasma membrane for cell to cell adhesion
and communication and relate their structure to function

• Identify and describe the modifications on the basal plasma membrane for cell to extracellular
matrix adhesion and relate their structure to function

• Identify and classify glandular tissue