…….is medicine an art or a science?

I think the answer lies somewhere in

the middle. It is both an art and a science. We use science to give us
some objective answers, drugs, tools, and to prove facts. The science of
medicine can be learned by anyone and requires only hard work and
perseverance.

It is the art of medicine, however, which has to be felt, loved, nurtured,

and finally accepted if we are to become
good clinicians.
…….science is much more than memorizing
pieces of information and regurgitating
them during an exam.

It’s about curiosity, it’s about exploration,

it’s about taking risks.
 …….remember that an expert is someone who knows
one more fact than you do. However, that one fact
may not be relevant to your case so you may be the
true expert!

3
 CELLS AND TISSUES:
Introduction to Histology and Cell
Biology

A.N Malik,
Department of Human Anatomy,
JKUAT
5 A.N Malik, Department of Human Anatomy, JKUAT. 02/25/2022
…..With the cell, biology discovered its atom . . . To characterize life, it was henceforth essential
to study the cell and analyze its structure: to single out the common denominators, necessary
for the life of every cell; alternatively, to identify differences associated with the performance
of special functions.

—François Jacob, La logique du vivant: une histoire de l’hérédité

(The Logic of Life: A History of Heredity), 1970

6 A.N Malik, Department of Human Anatomy, JKUAT. 02/25/2022

 What is histology

• Microscopic anatomy
• The study of the microanatomy
of:
• Cells
• Tissues
• Organs as seen through a microscope.

• It examines the correlation

between structure and function
(physiology and biochemistry).
7 A.N Malik, Department of Human Anatomy, JKUAT. 02/25/2022
R’ship btwn gross anatomy, histology and physiology

8 A.N Malik, Department of Human Anatomy, JKUAT. 02/25/2022

 Why study histology

Histology offers the meeting point for

the mainstream preclinical sciences:
biochemistry, gross anatomy and
physiology.

Form follow function, function follows

form?

Form follows function (Louis Sullivan,

All diseases are 1856-1924)
disturbances at the cellular
level (Rudolph Virchow,
1858).
9 A.N Malik, Department of Human Anatomy, JKUAT. 02/25/2022
Levels of organization (cell to organism)

02/25/2022

10 A.N Malik, Department of Human Anatomy, JKUAT.

 The tools of trade

Dimensions and resolution

11 A.N Malik, Department of Human Anatomy, JKUAT. 02/25/2022

Light and electron microscope photomicrographs comparison

12 A.N Malik, Department of Human Anatomy, JKUAT. 02/25/2022

 Tissue processing

A.N Malik, Department of Human Anatomy.

JKUAT
 Tissue processing

14 A.N Malik, Department of Human Anatomy. JKUAT 02/25/2022

 Internet resources for further
reading

• https://www.leicabiosystems.com/knowledge-
pathway/an-introduction-to-specimen-
processing/

• https://www.youtube.com/watch?
v=P0cZKCfyUwE

15 A.N Malik, Department of Human Anatomy. JKUAT 02/25/2022

Assignment
Read and make notes on….

The effect of tissue processing on specimens.

16 A.N Malik, Department of Human Anatomy, JKUAT. 02/25/2022

 Tissue staining
Creating contrast in tissues and cells to allow visualization
under a microscope

17
A.N Malik, Department of Human Anatomy, JKUAT. 02/25/2022
Why stain tissues? Unstained (A) and stained (B) sections
of the prostate
 Hematoxylin and Eosin stain overview
Hematoxylin and Eosin
• A routine stain made up of
stains to stain different parts
of the cell and ECM.

• Routine stain used in frozen

sections, FNA and paraffin wax
processed tissues.

• Demonstrates a wide range of

cytoplasmic, nuclear and ECM
features.
19 A.N Malik, Department of Human Anatomy, JKUAT. 02/25/2022
 Hematoxylin

• Sourced from Hematoylon

campechianum. Oxidation produces
Hematein which is the actual dye in
hematoxylin.
• Mordants improve anionic
attachments
• Illustrates nuclear detail in cells.
• Depth of coloration dependent on
amount of DNA and length of time
of exposure to stain
20 A.N Malik, Department of Human Anatomy, JKUAT. 02/25/2022
 Eosin
• A cytoplasmic stain
• Typically pink with different
shades of pink for the different CT
fibers.
• Addition of acetic acid sharpens
the eosin stain.

21 A.N Malik, Department of Human Anatomy, JKUAT. 02/25/2022

 Stain differentiation and blueing
• Differentiation: Selective removal of stain from a tissue.
• Hematoxylin is differentiated using HCl (rapid differentiation) or acetic
acid (slow/controlled differentiation).
• Blueing: changes hematoxylin from red to the traditional blue color we
see. Slightly basic solutions like Scott’s tap water are used. They
chemically alter the dye to produce the blue coloration of nuclear
material.

22 A.N Malik, Department of Human Anatomy, JKUAT. 02/25/2022

 H&E staining protocol

• There are 3 types of H&E staining

protocols
• Progressive
• Modified progressive
• Regressive

• Progressive
• Hematoxylin staining without
differentiation
• May end up staining charged sections
of tissue including non –cellular
material like mucin which can be an
indicator of well differentiated tumors.

23 A.N Malik, Department of Human Anatomy, JKUAT. 02/25/2022

H&E staining protocol
• Modified progressive and regressive

24 A.N Malik, Department of Human Anatomy, JKUAT. 02/25/2022

 H&E Staining protocol

25

A.N Malik, Department of Human Anatomy, JKUAT. 02/25/2022

 Colon H&E. Note the balanced
coloration in this section of colon.
The cilia on the columnar epithelium
are very crisp.
26
A.N Malik, Department of Human Anatomy, JKUAT. 02/25/2022
Next class….

The cell….

Thank you!
27 A.N Malik, Department of Human Anatomy, JKUAT. 02/25/2022
 THE CELL
CYTOLOGY

02/25/2022
28
 Overview
• The human body is organized into 4 basic
tissues(nervous, muscle, epithelial and connective) that
consists of cells and ECM.
• THE CELL is the basic structural and functional unit
of all living organisms.
• Human beings have nearly 60*1012 cells , some 200
cells of different sizes, shapes, location as well as
function.

29 A.N Malik, Department of Human Anatomy, JKUAT 02/25/2022

 Overview
• Protoplasm: cell nucleus(holds the genome)and
cytoplasm(aqueous gel made of 70% water, proteins,
lipids, carbohydrates inorganic and organic molecules).
• Organelles and inclusions are suspended in the
cytoplasm.
• Apart from a mature RBC all other cells poses a
nucleus that conforms to the shape of the cell. Most
cells are mononucleated, others are multinucleated e.g.
osteoclasts, skeletal muscle cells.

30 A.N Malik, Department of Human Anatomy, JKUAT 02/25/2022

 Overview
• A cell membrane encloses the cell whereas a nuclear
envelope encloses the nucleus.
• The internal structure of cells is usually modified to
reflect the function of that cell e.g. muscle cells
contract thus contain lots of contractile
proteins(actin and myosin) and lots of mitochondria
for energy production.

31 A.N Malik, Department of Human Anatomy, JKUAT 02/25/2022

General organization of the cell

32 A.N Malik, Department of Human Anatomy, JKUAT 02/25/2022

 General organization of the cell

33 A.N Malik, Department of Human Anatomy, JKUAT 02/25/2022

 Overview
• Organelles within the cell are categorized into
membranous or non-membranous.
• Membranous organelles include: rough and smooth
ER, golgi apparatus, plasma/cell membrane,
endosomes, lysosomes, transport vesicles,
mitochondria, peroxisomes, microtubules, centrioles,
ribosomes, filaments….

34 A.N Malik, Department of Human Anatomy, JKUAT 02/25/2022

 Light and electron microscopic
features of cell organelles

35 A.N Malik, Department of Human Anatomy, JKUAT 02/25/2022

 Light and electron microscopic
features of cell organelles

36 A.N Malik, Department of Human Anatomy, JKUAT 02/25/2022

 MEMBRANOUS
ORGANELLES

02/25/2022
37
PLASMA MEMBRANE
• A phospholipid bilayered
structure that makes the
outer boundaries of a cell.
• It is embedded with
integral and peripheral
proteins which fxn as:
• CAMs
• Transmembrane transport
• Cell recognition molecules
• Intercellular communication
38 A.N molecules
Malik, Department of Human Anatomy, JKUAT 02/25/2022
PLASMA MEMBRANE
Ultrastructure and fxn of cell membranes

• Membranes are semipermiable barriers

that selectively regulate movement of
substances in and out of the cell.

• Composition:
• 35% lipid
• 60% proteins
• 5% CHO.

• Membranes are beyond the resolution

limit of a light microscope thus not
visible without special techniques.

39 A.N Malik, Department of Human Anatomy, JKUAT 02/25/2022

PLASMA MEMBRANE
• In electron microscopy, membranes
have a trilaminar appearance i.e. 2
dark lines separated by a thin
electron lucent zone. The unit
membrane is 5-8nm thick.
• Membranes are made up of a lipid
bilayer with a highly dynamic fluid
mosaic model because of the
cholesterol component.
• 2 hydrophobic phospholipid leaflets
face inwards and the hydrophilic
polar heads face outwards.
40 A.N Malik, Department of Human Anatomy, JKUAT 02/25/2022
PLASMA MEMBRANE
• Lipid rafts: these are localized
areas within the cell membrane
that contain high amounts of
cholesterol and glycosphyngolipids.

• Lipid rafts are thicker and less

fluidy and offer signaling
platforms i.e. integral and
peripheral proteins used in
signaling are abundant hear.

41 A.N Malik, Department of Human Anatomy, JKUAT 02/25/2022

PLASMA MEMBRANE
Membrane proteins

• There are two types of membrane

proteins based on their spatial
orientation on the cell membrane i.e.

• Integral/intrinsic proteins: traverse the

whole thickness of the cell membrane.
• Peripheral/extrinsic: found on either
side of the cell membrane i.e.
cytoplasmic or extracellular

• Functions of membrane proteins: see

earlier slides.
42 A.N Malik, Department of Human Anatomy, JKUAT 02/25/2022
 PLASMA MEMBRANE

43 A.N Malik, Department of Human Anatomy, JKUAT 02/25/2022

PLASMA MEMBRANE
Membrane proteins
Categorized into 6 based
on their function i.e.
• Channels
• Pumps
• Receptors
• Linkers
• Enzymes
• Structural proteins
The categories are not mutually
44 A.N exclusive.
Malik, Department of Human Anatomy, JKUAT 02/25/2022
 PLASMA MEMBRANE
Membrane proteins
• Pumps: act as transporters of substances across the
cell membrane e.g. ions, a.a, sugar either with
linkage to sodium pump or singly by themselves.
Pumps utilize energy.
• Channels: allow passage of substances across the cell
membrane via passive diffusion. No energy required
but just a concentration gradient.

45 A.N Malik, Department of Human Anatomy, JKUAT 02/25/2022

 PLASMA MEMBRANE
Membrane proteins
• Receptors: act as recogntion molecules and localized
binding ligands in hormonal stimulation, antibody
reactions and coated vesicle endocytosis.
• Linker proteins: they are anchorage molecules. They
link the cytoskeleton to the ECM e.g. integrins link
actin to fibronectin.
• Enzymes: play various roles e.g. ATPases, ATP
synthase, dipeptidases….

46 A.N Malik, Department of Human Anatomy, JKUAT 02/25/2022

 PLASMA MEMBRANE
Membrane transport and vesicular transport
• Transport across the cell membrane can either be thro
simple diffusion or membrane transport proteins.
• 2 major classes of transport proteins
• Carrier proteins: highly selective. They bind a molecule and
undergo conformational changes to release the bound molecule
to the other side of the membrane. May(active) or may not use
energy(passive).
• Channel proteins: transmembrane proteins with hydrophilic
proteins(ligand, mechanically or voltage gated)

47 A.N Malik, Department of Human Anatomy, JKUAT 02/25/2022

 PLASMA MEMBRANE: membrane
transport

48 A.N Malik, Department of Human Anatomy, JKUAT 02/25/2022

PLASMA MEMBRANE: membrane
transport
Vesicular transport
• Localized configurational changes
on the cell membrane occur and
vesicles form or fuse with it.
Offers transport for large
molecules.
Types of vesicular transport
• Endocytosis: substances enter the cell
• Exocytosis: substances leave the cell

49 A.N Malik, Department of Human Anatomy, JKUAT 02/25/2022

PLASMA MEMBRANE: membrane
transport, endocytosis

Phagocytosis
50 A.N Malik, Department of Human Anatomy, JKUAT 02/25/2022
 PLASMA MEMBRANE: membrane
transport, exocytosis

• Exocytosis transports molecules from the cell to the

extracellular space. There are two major pathways
involved:
• Constitutive pathway: substances secreted leave the cell
immediately e.g. antibodies
• Regulated secretory pathway: products to be secreted are
stored in secretory vesicles in the cytoplasm until a stimulus
for their secretion is given e.g. hormones. Calcium is required
for the secretory vesicle to fuse with the cell membrane
Read and make notes on precise targeting and specificity in
vesicular transport.

51 A.N Malik, Department of Human Anatomy, JKUAT 02/25/2022

 Internet resources for the cell membrane

• https://www.ncbi.nlm.nih.gov/books/NBK9898/

52
A.N Malik, Department of Human Anatomy, JKUAT 02/25/2022