MICROSCOPE

Light Microscope Transmission Electron Microscope Scanning Electron Microscopes

(max 1600x) (TEM) (max 1500000x) (SEM)
(max 200000x)

light passes through a electron beams pass through a thin electron beams scan over the surface of
specimen or a thin slide slide of a specimen to form an image a specimen to form an image
of it to form an image

colored black and white 2d internal black and white 2d external

observe living magnifications and resolution of the /

specimens, can be images produced are higher, so more
prepared easily details can be seen

- eyepiece: magnifying lens we look through, with 5x, 10x, 16x times
- objective: magnifying lens pointing to the specimen
- condenser: focuses light onto the specimen; diaphragm: adjust amount of light
- coarse/fine adjustment: we turn it to raise or lower the stage to get a certain focus, small/large
movement
- magnification: size of image/ size of object

- procedure:
1. Sectioning: allow light to pass through
2. Staining: make cellular structures more distinct
3. Mounting: provide a flat surface for observation or to prevent the specimen from drying up or to
protect the objective lens from getting dirty.

CELL STRUCTURE
1. nucleus: spherical structure bounded by nuclear
membrane,
a. pores to allow exchange of materials,
contains DNA that carries genetic
information, and a nucleolus
b. synthesise rna/dna, site of transcription,
control cell division and movement (rbc no
nucleus, no division of cells)

2. cytoplasm: jelly-like fluid consisting of water and

proteins,
a. holds organelles,
b. site for chem reactions
 3. cell membrane: thin and flexible membrane made of phospholipids and proteins, encloses the cell
and separates the cell contents from the outside environment,
a. differentially permeable, controls movement of substances in and out of the cell,
b. dark line (head of phospholipid) and light middle (fatty acid tail)

4. cell wall: thick and rigid outermost layer (cellulose), fully permeable, protects and gives shape

5. endoplasmic reticulum: network of interconnected membrane-bounded sacs, continuous with

the outer nuclear membrane and extends throughout the cytoplasm,
a. rough: has ribosomes attached, a site for the synthesis of proteins
b. smooth: no ribosomes attached, a site for the synthesis of lipids
c. proteins transported in vesicles (protein bound structure) pinched off from rough er +
golgi apparatus (allow membrane to invaginate and carry out endocytosis) to form cell
membrane

6. mitochondrion: double membrane highly folded -

a. increase surface area for enzymes to stick to catalyse the breakdown of food through
respiration to release energy,
b. main site of respiration (convert chemical energy in food into energy), more energy
requirement, more mitochondria (eg. synthesis of enzymes, absorption of nutrients and
minerals, contraction or high level metabolic activities)

7. chloroplast: double membrane, chlorophyll (green pigment) captures light energy and converts
into chemical energy in photosynthesis, starch grains are present inside

8. vacuole: fluid-filled sac bounded by a single membrane, contain water, enzymes and food in
animal cells; contain cell sap (water and dissolved substances), plant cells become turgid when
the large central vacuole is full of water, providing support to the plant.

prokaryotic cells eukaryotic cells

cell wall may be present cell wall present in plant, fungi, algae

no membrane, no true nucleus membrane bound, distinct nucleus

DNA lying free in cytoplasm relatively larger

FLUID MOSAIC MODEL

// Mosaic: transmembrane protein: some proteins interspersed in
phospholipid bilayer, held together with weak hydrophobic intersections,
provide channels/carriers for transporting polar molecules across the
membrane.
- Lateral movement, but not flip-flop
 - membrane is not rigid, can break and fuse readily, change shape size slightly as water
content changes during growth/cell division
- allow new vesicles to form from cell membrane, vesicles fuse w/ cell membrane

1. Endocytosis
- For bulk transport of substances which is too large to move across
- golgi apparatus

2. Exocytosis
- to allow membrane fusion, thus supporting export of materials contained in vesicles by
exocytosis & intracellular vesicular transport of materials

- Transmembrane proteins: act as antigens (glycoprotein) for recognition by other cells (ie allowing
cells to distinguish identity of neighbouring cells
- recognition: signaling molecules act as receptors for binding specific chemical
messenger (eg. hormones, neurotransmitters → turn on cell activity, relax message to
inside of cells)
Carbohydrates: antigens/recognition markers on the membrane surface for recognition by other cells.

FORMS OF TRANSPORT
a) Diffusion
Diffusion down the concentration gradient - net movement of particles down the concentration gradient
with no energy input, will stop when equilibrium is reached, molecules of a substance will continue to
move but stay equal, the steeper the concentration gradient, higher the rate, direction.
- Factors:
- smaller size of particles
- higher temperature: movement of phospholipid molecules in the cell membrane →
less packed
- gas rate > liquid rate
- larger surface area
- lipid soluble > water soluble
1. Simple Diffusion: Phospholipid bilayer: simple diffusion (oxygen and carbon dioxide pass
through)
a. Dissolve and Diffuse: cell membrane is permeable to hydrophobic molecules
b. Bilayer is a barrier to water molecules, ion and water-soluble molecules
2. Facilitated diffusion: assisted by protein channel/carrier proteins (not lipid soluble), depends on
concentration gradient, in phospholipid bilayer
a. Carrier proteins: allowing certain substances (hydrophilic substances) to pass through,
contributes to differential permeability of cell membrane, pass through against
concentration gradient by active transport and change shape.
b. antigen: glycoprotein, identify bad vs good cells, trigger immune response
c. receptor: bind to substance (eg hormones, antigens, enzymes), turn on cell activities
 d. enzymes: active site towards outside substances, in cell membrane, can bind and catalyse
chemical reactions

b) Osmosis
Movement of water molecules only, a kind of diffusion, move from high water potential to low water
potential, only take place through a selectively permeable membrane
- Water potential: tendency of water to move from one area to another, lowered if solute present
- Must answer: The water potential of region B is higher than that of region A, there is a net
movement of water molecules from region B to region A down the water potential gradient across
a selectively permeable membrane by osmosis
- Dialysis tubing: more obvious change in liquid level
- Hydrostatic pressure: develops due to weight of column of sucrose solution if rate of outflow
of water = rate of inflow of water, the column of sucrose solution stop rising, it balances the
tendency of water to enter due to present of solute to reach same water potential → no net
movement. BUT in control, water moves out the dialysis tubing slowly due to hydrostatic
pressure
- Action of cell:
- In hypertonic solution: water potential of the cell sap of cell A is greater than that of
sucrose solution → net movement of water molecules from cell sap of cell A to
sucrose solution down the water potential gradient across the differentially permeable
cell membrane by osmotion → cell (vacuole) shrink, pulls cytoplasm and cell
membrane away from the cell wall → plasmolysed and flaccid
- In hypotonic solution: cell swells and bursts, turgid
c) Active Transport
The movement of a substance against the concentration gradient (region of lower concentration →
higher concentration) Cannot take place in a dead cell (no respiration, no source of energy),
independent of concentration gradient.
- Substance to be transported binds with carrier protein → energy is used to change shape →
substance is released to other side → carrier protein resumes to its original shape
- Pass through by phagocytosis (cell eating), to take in large particles or cells: part of cell
membrane infolds and engulfs the particle, the membrane of the cup-shaped cavity fuses
and pinches off the form vesicle
- Biological importance: active uptake of mineral salts from soil by root epidermal cells which
have lower salt concentration, absorption of digested food in the small intestine.

ENZYMES
// Catabolism (complex to simple) / Anabolism (building up)
- Enzyme (biological catalysts) lower the energy barrier so reactions can take place at body temp
- at body temp, the metabolic rate is too low to support life if enzyme is absent) →
catalyses the reaction and increases rate of metabolic reactions (make “unstable”)
 1. Nature and properties
- Nature: Made up of globular proteins, bonding holds polypeptide chain together
- 3d shape maintained by weak bonding, can be broken → change in shape of active site →
denatured
- Highly specific, can usually catalyse one type of reactions only with a complementary substrate
- Only work efficiently within a particular range of temperature and pH
- At low temperatures, enzymes are inactive because the kinetic energy of enzymes and
substrate molecules is low (1) as temp increase, both enzymes and substrate have more
kinetic energy thus the chance of forming enzyme-substrate complex increase (1), as the
temp keeps increasing, high temperature cause denaturation of enzymes in their active
site so the substrate no longer fits the active site of enzyme (1).

2. Action
- transition state: unstable points with substrates have to pass through before converting into
products, energy is required to break or weaken the bonds in the substrate to reach the transition
state.
- (p5) at active site, substrate and enzyme’s shapes fit together, enzyme-substrate complex,
products can be reused (effective in very small amount) & enzymes are unchanged

3. Lock and key hypothesis - lock = substrate; key = enzyme

Demonstration of the catalytic action of enzymes

dv: catalase activity (high/low)
indicated by: rate of oxygen production (fast/slow)
measured by: volume of gas collected per unit time calculated from the volume of gas collected in a fixed
period of time.

inverted pipette, reacting mixture is drawn into it, measure initial liquid level, drop in liquid level in the
pipette in a fixed period of time

CELL DIVISION
1. Chromosomes
- dna molecules coiling around proteins
● when cell is not dividing: chromatin (invisible/fibre)
○ dispersed = allows DNA to be replicated,
○ genetic information carried in the DNA can be read,
○ protein can be synthesized
● when cell division begins: chromosomes (visible under light microscope = thread like)
○ coil up tightly and become shorter and thicker
○ chromosomes can be moved into the daughter cells more easily without tangling during
cell division
○ chromosome is in duplicated state = two identical chromatids (sister chromatids) joined
at centromere, each chromatid contains one DNA molecule
● during cell division:
○ sister chromatids separate and each goes into one daughter cell
● after cell division:
○ chromosomes uncoil and exist as a mass of chromatin fibres again

- homologous chromosomes
● 23 pairs of homologous chromosomes in a human cell (ie 46 chromosomes)
○ in each pair, one is paternal chromosome, the other is maternal chromosome
○ diploid: contains two sets of chromosomes - both members of each homologous pair of
chromosomes in each cell (eg. somatic cells)
○ haploid: one set of chromosomes - one member from each homologous pair (eg.
gametes/sex cells)

2. Somatic cell division

- somatic cells can divide by mitotic cell division to form new cells
- mitotic cell division (bc same as parent cell)
- growth: new cells for multicellular organisms
- repair: replacing/repairing worn out cells
- asexual reproduction: produce offspring that is genetically identical
● interphase (cell growth, 90% of the time)
○ cell carries out its functions within the organism and prepares for cell division:
○ it synthesizes proteins and produces new organelles
○ grows to its maximum size
○ carries out DNA replication to form two daughter cells that are identical to the parent cell
(same no. and kinds of chromosomes)
● mitosis (nuclear division in body cells)
○ separation of the sister chromatids,
○ equal distribution of the separated chromatids between the 2 daughter nuclei
○ 4 phases
1. prophase
a. chromosomes shorten and thicken, become visible
b. nuclear membrane breaks down
2. metaphase
a. chromosomes line up at the equator
b. spindle fibres attach to chromosomes
3. anaphase
a. spindle fibres contract, sister chromatids separate
and move to opposite poles of the cell
b. cytoplasm starts to divide
4. telophase
a. a new nuclear membrane forms around each group of
chromosomes
b. the chromosomes uncoil and become chromatin again
● cytokenesis (cytoplasmic division)
 ○ cytoplasm of the parent cell divides into 2 equal halves, forming two daughter cells
1. animal cells: cell membrane constricts inwards until the cell splits into two
2. plant cells: cell plate made up of new cell membranes and cell walls is formed
between two daughter nuclei → cell plate grows outwards, dividing the cell
into 2.

3. Gametes cell division

- gamete-producing cells carry out meiotic cell division to produce haploid gametes
- a cell divides to form 4 haploid daughter cells, which are genetically different from
parent cell and from each other, no DNA replication occurs.
- meiotic cell division:
- produce hapoid gametes: sexual reproduction, when male and female gamete fuse,
diploid no. of chromosomes is restored, so it can be restored and preserved
- produce genetic variation in gametes
- independent assortment of chromosomes and crossing over = diff. in genetic
materials
- increase chance for the species to survive when environment changes
1. independent assortment of chromosomes:
a. in metaphase 1, pairs of homologous chromosomes line up randomly
along the equator of the cell (maternal/paternal on which side is
random) → various genetic combinations
2. crossing over
a. crossing over: exchange between non sister chromatids of homologous
chromosomes. matching regions on non-sister chromatids break,
exchange and reconnect to the chromosome.
b. brings about/results in variation in gametes and gives rise to new genetic
combinations, allowing survival of the fittest to occur when
environmental condition changes
c. chiasmata: cross-shaped structure formed when a chromatid from each
homologous chromosome wraps around each other
● first meiotic division
○ pairing and separation of homologous chromosomes (ie divide by half)
○ 4 phases
1. prophase 1
a. chromosomes shorten and thicken, become visible
b. homologous chromosomes pair up
c. crossing over
d. nuclear membrane breaks down
2. metaphase 1
a. homologous pairs line up along the equator
b. spindle fibres attach to the chromosomes
3. anaphase 1
 a. spindle fibres contract, two members of each homologous pair separate
and move to opposite poles of the cell
b. cytoplasm starts to divide
4. telophase 1
a. a new nuclear membrane forms around each group of chromosomes
● cytokenesis (cytoplasmic division)
○ two haploid daughter cells with duplicated chromosomes are formed
● second meiotic division
○ separation of sister chromatids of each chromosomes (ie mitotic but w/ haploid)
○ 4 phases
1. prophase 2
a. nuclear membranes break down again
2. metaphase 2
a. chromosomes line up along the equator
b. new spindle fibres attach to the chromosomes
3. anaphase 2
a. spindle fibres contract, sister chromatids separate and move to opposite
poles of the cells
b. cytoplasm starts to divide
4. telophase 2
a. new nuclear membrane forms around each group of chromosomes
b. the chromosomes uncoil to become chromatin again
● cytokenesis: four haploid daughter cells are formed

experiment:
- subjectivity: different groups used slightly different criteria to categorise the stages and
different groups collected their data from different samples
- creativity: inferences are a product of human creativity
- the categorisations of stages is also tentative, it may change in the future as more knowledge is
shared
- tentativeness: the conclusion may change with additional data, different samples or
different criteria for determining the stages of cell cycle
- observation vs inference: using observation to make inferences, no actual timing of the
different stages has ever been made
- another method to reduce error: choose the area to be observed in a random way instead of
focusing on an area where lots of cells are undergoing mitosis; random sampling can reduce
biased sample
- systematic error: using a stretched measuring tape (solve = calibration)
- random: rounding values up, people take turns for reading (solve = take average)

Test feedback 9.5/30 (avg: 16)

1. Photomicrograph
 a. Mitotic cell division (1). Root tip is not the reproductive part of the plant (1).
b. Cell
i. Label cell at interphase
ii. replication of DNA/duplication of chromosomes (1). Synthesis of organelles (1)
such as mitochondria/ER etc.
c. Arrangement
d. Describe
i. Chromatin condenses to form chromosomes (1). The nuclear membrane breaks
down (1)
ii. The chromosomes line up in the middle of the cell (1). Spindle fibres attach to
the chromosome (1)
e. Explain difference in number of chromosomes
i. The number of chromosomes in stage 2 is half of that in stage 1 (1), it is because
the sister chromatids of each chromosome in stage 2 are separated from one
another thus the number of chromosomes doubled (1).
ii. It ensures that the daughter cells formed are genetically identical to the parent
cell (1) which is important for growth of the organism/asexual reproduction
f. In plant cells, cytoplasmic division involves the formation of a cell plate in the equatorial
region of the cell while animal cells do not (1). Cytoplasmic division is accomplished by
the formation of a cleavage furrow/ inward construction of cell membrane around the
middle of animal cells but not in plant cells (1)
g. Graph: dna content at 1 at interphase and increases to 2 at mitosis, then drops in the
middle of mitosis and cytokinesis, then lands at 1 in cytokinesis
2. Ovum
a. Title, cell outline, draw 3 chromosomes with different length, shown which are single
threads
b. The pairs of homologous chromosomes segregate into the daughter nuclei during the first
mitotic cell division (1). After separation, the daughter cells formed would have only one
member of each pair/haploid (1). The sister chromatids of each chromosome then
separate in the second meiotic cell division (1)
c. Crossing over occurs between non-sister chromatids of homologous chromosomes (1),
resulting in formation of new combinations of genetic materials carried by chromosomes
in the ovum (1). Independent assortment of homologous chromosomes (1) resulting in
different combination of chromosomes in the ovum (1)
TEST FEEDBACK
Test 1 (ch 1, 2, 5): 15/30 (avg: 15.9)
1. Experiment analysis
a. The concentration of glucose of the same sample measured in different trials and by
different students are not the same (1). By repeated measurement and taking average,
random errors can be reduced (1). Therefore, a value closer to the true value can be found
and reliability of results increases (1).
b. Add equal volume of Benedict’s solution to the orange juice (1). Heat the mixture in a
boiling water bath (1). Filter the mixture with filter paper (1). Dry the precipitate on the
filter paper in an oven until a constant mass is obtained (1). Weigh the mass of the brick-
red precipitate. (1)
c. The oxygen in air will oxidize the vitamin C in the orange juice (1) and lose its reducing
property (1). Therefore, the vitamin C concentration measured will be underestimated (1)
d. Glucose is a reducing sugar (1) which will also reduce the DCPIP solution (1). Therefore,
the presence of glucose in the orange juice will lead to overestimation of the vitamin C
concentration measured in this experiment (1).
2. Food substances
a. (e) (ii) Proteins are built from 20 different amino acids (1) joined together in different
sequences to form polypeptide (1) which can coil/ fold up to different shapes (1).
b. (g) We can obtain sufficient vitamin D by exposing skin to sunlight (1). Vitamin D can
only help the absorption of calcium, not vitamin D, is the raw material to strengthen the
bone, preventing rickets.

Test 2 (ch 3, 4, 11): 14/30 (avg: 13.1)

1. Phospholipids function and structure
a. molecules Y allows lipid soluble substances to dissolve and diffuse across it (1).
However, the hydrophobic tail of molecules Y forms a hydrophobic core of cell
membrane (1) that repels polar or charged molecules (1), thus these molecules can only
be transported by molecules Z which acts as channels for them (1). Molecules Y are
tightly packed together, so large molecules cannot pass through the spaces between them
(1).
b. Arranged in a bilayer in cell membrane (1) with the hydrophilic heads of phospholipids
point outwards, hydrophobic tails point inwards (1). It is because the medium inside and
medium outside the cell are aqueous solutions (1). Hence, only the hydrophilic part of the
phospholipids is in contact with the cell sap and extracellular fluid (1).
c. Carbohydrates (1). It acts as antigens/ recognition markers on the membrane surface for
recognition by other cells (1).
2. Plant cells
a. A: Nucleolus B: Chloroplast C: cell membrane/cell wall D: nuclear membrane/nucleus
b. magnification: 2.9 x 10^4 cm
c. Resolution is not high enough (1) to view this fine structure (1).
d. Resemblance of cell *add cell wall, title
 e. The internal structure of chloroplasts cannot be seen under light microscope (1) showing
that the advances of microscopy enables scientists to discover the structure of cell
organelles (1), hence, the study about the functions of organelles is feasible.

exam skills:
- increase sample size = increase reliability, not accuracy (for accuracy, consider using equipment
with smaller division or controlling more variables)
- no “pass through”, must use dissolve and diffuse
- describe first, then explain.
- no vague terms like “stop increasing”, instead “remains unchanged”
- no “hypotonic”, only compare water potential; cells cannot be hypertonic/hypotonic, only the
solution.
- water potential gradient vs concentration gradient

Pop-Up Test
1. Osmosis experiment
a. Graph (liquid level acceptable, no water level)
b. The level of sucrose solution in the capillary tube increases (1) and then remains
unchanged (1). It is because the water potential of the sucrose solution is lower than that
of the distilled water (1) so there is a net movement of water molecules from the distilled
water to the sucrose solution across the selectively permeable cell membrane of the fish
swim bladder tissue down the water potential gradient by osmosis (1). It then stops rising
and remains unchanged as the hydrostatic pressure developed by the weight of the liquid
column balances the force developed by the difference in the concentration of solution
inside and outside the tubing (1).
c. The rate of increase of solution level with 10% glucose solution is lower than that with
10% sucrose solution (1) because glucose could diffuse out of the fish swim bladder
while sucrose cannot (1). The water potential gradient between water and 10% glucose
solution is less steep than that between water and 10% sucrose solution (1). Thus the rate
of osmosis is lower for 10% glucose solution than 10% sucrose solution. (1).
d. (i) The same set-up (1) using distilled water to replace the 10% sucrose solution (1)
(ii) The rate of osmosis can be calculated by calculating the sum of the rise in solution
level in the experimental set-up and the fall of the solution level in the control set-up (1)
per unit time (1).