Cell and Microscope
Cell and Microscope
Cell and Microscope
Cell
Robert Hooke named “cell” in his book “Micrographia”
after observing cork under a lense
Anton van Leeuwenhoek A master microscope maker identified “Animalcules”
Robert Brown:
• noticed that pollen grains in water jiggled around called “ Brownian motion”
• And discovered the nucleus
Louis Pasteur:
• Disproves the prevailing theory of “Spontaneous generation” with his swan‐neck
flask experiment >> Populations need to be seeded by existing population
• The discovery led to invention of pasteurization and autoclave
Swan‐neck experiment
Cell Theory
• Schielden and Schwan
Cells are the building blocks of structure in living things.
Cells are the smallest unit of life, All organisms are made
of one or more cells.
• Rudolf Virchow
Cells are made from other cells (pre‐existing cells) by cell division.
To this we can now confidently add two concepts.
• Cells contain a blueprint (information) for their growth, development and behavior.
• Cells are the site of all the chemical reactions of life (metabolism).
Endosymbiosis theory
- -
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-70s
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•
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ribosomes
Why cells have to divide and why
they need circulatory system?
Microscopy
A microscope is used to produce a magnified image of an object or specimen. Today, cells
can be observed by two fundamentally different types of microscopy:
Cell fractionation
ocular lens no
/ 1in eyepiece)
Microscope
Examining the structure of living cells
Different depth of field
→ adjust with file adjustment knob
Recording observations
What you see with a compound microscope may be recorded by drawings of various types.
For a clear, simple drawing:
• use a sharp HB pencil and a clean eraser
• use unlined paper and a separate sheet for each specimen you record
• draw clear, sharp outlines and avoiding shading or colouring
• use most of the available space to show all the features observed in the specimen
• label each sheet or drawing with the species, conditions (living or stained), transverse section (TS)
or longitudinal section (LS), and so forth
• label your drawing fully, with labels positioned clear of the structures shown, remembering that
label lines should not cross
• annotate (add notes about function, role or development), if appropriate
• include a statement of the magnification under which the specimen has been observed
Example of drawing
Example of drawing
Magnification
Magnification
• Ratio of image size to actual size
• The number of times larger an image is than the real size of the object
Total magnification
Magnification is product of objective lens and ocular lens
Magnification without resolution
Resolving power (Resolution)
↳• Ability to separate or distinguish between two objects that are very close together
as separate; what allows us to see detail
• The higher the resolution of an image, the greater the detail that can be seen
If two points can be resolved, they will be seen as two points
-
electromagnetic spectrum
How do we see thing?
• We see objects because light from a luminous object reflects off of the object
• Some of the colours of light are absorbed
• Only the colours that are not absorbed are reflected and enters our eyes.
• Limit of resolution is about one half the wavelength of
the radiation
• The best resolution that can be obtained using a
microscope that use visible light is 200 nm
• If the object is any smaller than half the wavelength
of the radiation, it cannot be seen separately from
nearby objects
• The corresponding maximum useful magnification of
about 1500 times
• To increase resolution:
• electrons are used (free electron behave like
electromagnetic radiation)
• very short wavelength: resolution of electron
microscope is 0.5 nm
• negatively charged: be focused easily using
electromagnetic coil
Electron microscopy‐ the discovery of cell ultrastructure
magnetic
[ nucleus
Transmission electron microscope
Mechanism of electron
microscope
Limitations of the electron microscope – and how these
are overcome
1 Electrons cannot penetrate materials as well as light does.
• Specimens must be extremely thin for the electron beam to
penetrate and for some of the electrons to pass through.
• sliced into very thin sections using a special machine called a
microtome
• stained with heavy metal ions (such as lead or osmium) to
make them absorb electrons at all
2 Air inside the microscope would deflect the electrons and destroy
the beam.
• The interior of the microscope must be under a vacuum
• no living specimens can survive inside the electron
microscope when in use.
• Water in cells would boil away in a vacuum. Sections are
completely dehydrated, so given that cells are 80–90 per
cent water
Electron micrographs of Liver cell
Electron micrographs of nucleus
Light microscope VS Electron microscope
Calculating magnification
We might want to know how many times an image has been magnified
The scale bar represents the “real” size of the sample in the image, so we only need to work with the
scale bar
Scale bar is the line added to micrograph or drawing to
help to show the actual size of the structure.
stage micrometer
↑
division
"
"
super -
imposed
-
**Units for size
measurements
• cm
• mm
• µm
• nm
Calculating actual size from magnification
Calculating actual size from scale bar
Questions:
What is the magnification?
How long is one of the rust‐coloured bacteria
What is the size of the neutrophil, at the
widest point?
Ultrastructure
The fine (detailed) structure
of a cell revealed by electron
microscope
Animal cell Plant cell
↓ ↳
containing centrioles .
contains centrosomes
without centrioles
Nucleus and Nucleolus compacted area of condensed
/
chromatinin , produce ribosomes
Largest organelle
Double membrane: Nuclear envelope
Outer membrane connected with ER
Nuclear pore
Hierarchical structure of
chromosome
Ribosome
• 25 nm in size
• Unbound organelle
• Compost of Proteins and rRNA
• Two subunits: large and small subunits
mmmm
• Function: protein synthesis (translation)
Types of Ribosome classified by size
Types of Ribosome
classified by position
*
Ribosome assembly in Nucleolus
Endoplasmic reticulum
Endoplasmic reticulum (ER)
• An interconnected network of flattened,
membrane‐enclosed sacs or tubes (Cisternae)
• Two types of ER
• Rough ER; rER (Ribosomes attached on)
function of rER: fold protein in correct shape
• Smooth ER; sER
Function of sER:
• Lipid synthesis (fatty acids, cholesterol and
cells
steroid hormones / testes→
, ovaries .
liver cells
• Detoxification of toxic substances
→
• Store Calcium ion in muscle for muscle
Muscle cells
contraction
.
• Glycogen synthesis
Protein folding in rER
rER provides separate chemical
environment which allow correct
protein folding
Formation of membrane protein on rER membrane
Role of Sacroplasmic reticulum in muscle contraction
Golgi body/ apparatus/ complex
Cis face‐ “receiving side of
Golgi apparatus”
• Stack of flattened sacs + Golgi vesicle
• Golgi vesicles form at the ends of each sacs
Pink aging
• Function: collecting, modifying, sorting and
transporting of proteins
• Protein processing:
• adding sugar chains into protein to form
Glycoprotein
• Cleave Methionine from newly formed
Trans face‐ “shipping side of protein to form functioning protein
Golgi apparatus” • Convert sugar in to cell wall in plant
lysosome
-
↑
golgi
vehicle →
lysosome
I
mitochondria
• 0.1‐0.5 micrometer in diameter
• Spherical sac contains hydrolytic enzyme
• Function:
• breakdown of unwanted structure, old organelles or cells
• Digest engulfed bacteria
• Replacement of cartilage with bone
• Acrosome, lysosome in head of sperm,: digest a path to ovum
Endomembrane system
The system is a group of membranes and
organelles that works together to modify,
package and transport lipids and proteins
Component
• Nuclear envelope
• Endoplasmic reticulum modify
1) collect
↓ 3)
2)
• Golgi apparatus packaging
↑
sorting
• Vacuoles
4)
• Lysosomes
• Plasma membrane
These components are either continuous or
connected through transfer by vesicles
Vesicular transport
Cytoskeleton
Motor protein with vesicle for
vesicular transport
Mitochondrion/Mitochondria
• One micrometer in diameter and vary in shape
• Double membrane (inner and outer membrane)
• Own protein synthesizing machinery, 70s
Ribosomes and mitochondrial DNA
• Structures:
• Porin (transport protein)
• Cristae
• Matrix
Function of mitochondrion
Main function: aerobic respiration
Transfer energy from energy‐rich molecules
to ATP, energy‐carrying molecule
Other function: synthesis of lipids during
respiration
Breaking down and synthesis of ATP
Chemical reactions in Mitochondrion
Arrangement of microtubules in Flagellum and Cilia
How flagellum and cilia move in motion
Centrosome
(Microtubule organizing center, MTOC)
MTOC is an area where microtubules are produced
Function: organizing the assembly of spindle fibre
during cell division, formation of cytoskeleton, cell
signaling pathway
Present only in animal, not in plant Centriole: Nine triplets of
• In animal, centrosome = pair of centrioles, each microtubules arranged in a ring
made up of a ring of nine groups of microtubules
Formation of microtubule
Microtubules during cell division
Cell membrane or Plasma membrane
• 7 nm across
• It appears as Double line on EM “Trilaminar appearance”
• Function: control exchange between the cell and environment
Microvilli (Finger‐like extension of cell membrane)
On epithelial cell of gut Lining of proximal convoluted tubule
Function of membrane protein
Transportation through membrane
Chloroplast
• 3‐10 micrometer, elongated shape
• Double membrane
• Own protein synthesizing machinery, 70s Ribosomes and
chloroplast DNA
• Contain many pigments and electron carriers on thylakoid
membrane to trap light energy to make ATP
• Thylakoid: fluid‐filled sac
Types of plastid
Leukoplast : store starch
Chloroplast
Chromoplast : pigment
synthesis and storage
Light‐dependent reaction
The Light‐dependent and light‐independent reaction
Cell wall and
Plasmodesmata
Central Vacuole (Sap vacuole)
Large and permanent in plant cell
Function:
• store nutrients and/or waste products
• regulate osmotic properties of cells
Cell sap contains water, sugars, mineral salts,
lipids, proteins, starch, pigments, enzymes
Tonoplast is the membrane that surrounds the
central or large vacuole of the plant
• Provide pressure (turgor pressure) to
maintain cell structure
Contractile vacuole
Food vacuole
Bacteria
Bacteria
• they are unicellular
• typically 1–5 μm in diameter
• cell walls made of peptidoglycan, composed of
polysaccharides and peptides combined together
• lack organelles surrounded by a double
membrane in their cytoplasm
• have a single circular chromosome that is ‘naked’
(of DNA without associated proteins)
• ribosomes are present, but they are the smaller
70S variety.
Bacterial conjugation (Sexual reproduction of bacteria)
Type of bacteria
Difference between Gram positive and negative bacterial cell wall structure
Gram staining
*Stain will be fixed due to formation of a complex of crystal violet and KI
Prokaryotic and eukaryotic cells compared
The fundamental differences in size and complexity of prokaryotic and eukaryotic cells are highlighted in
Prokaryotes, e.g. bacteria, cyanobacteria Eukaryotes, e.g. mammals, green plants, fungi
cells are extremely small, typically about 1–5 µm in diameter cells are larger, typically 50–150 µm
nucleus absent: circular DNA helix in the cytoplasm, DNA not nucleus has distinct nuclear envelope (with pores), with
supported by histone protein chromosomes of linear DNA helix supported by histone protein