Guide To STPM Practicals

Oxford Fajar Sdn. Bhd.
(008974-T) 2012 1
A GENERAL GUIDE TO PRACTICAL WORK
A. Specimen Drawings
1. Drawings should be done with a mechanical pencil with a 2B lead.
2. Drawings should be as large as possible between half to three-quarters of a page on blank sheets
of paper.
Large drawing() Drawing too small ( X )
3. Outline of drawings should be clear, clean and continuous to show that the specimen drawn is
functional. It should not be sketchy.
Drawing continuous line () Drawing line broken ( X ) Sketchy drawing (X)
4. The overall drawing should be accurate, proportional and two-dimensional. Shading of any portion
of the drawing to show depth is not allowed but dots and slashes could be used if necessary.
Drawing dots or slashes to indicate depth ()
5. Drawings should be labelled as far as possible and done outside the drawings. Different parts of
the specimen are indicated by label lines which should not be seen crossing each other as shown
below. All labelling should be done in pencil.
T.S. of monocotyledon root T.S. of dicotyledon root
6. Magnification of drawings should also be estimated and stated.
Size of drawing
Magnification of drawing = (= for example, 3x)
Actual size of specimen
Guide to STPM Practicals
2 Oxford Fajar Sdn. Bhd. (008974-T) 2012
or
Magnification of eyepiece Magnification of objective Size of drawing
Magnification of drawing =
Apparent size of specimen
7. In the case of microscopic specimens, two types of drawing can be done :
(i) plan drawing which is done when observing the specimen under the low or medium power
objective of the microscope to see the overview of the specimen. Outline of the specimen is
drawn which may contain layers to indicate the distribution of various tissues without
showing any cells
(iii) detailed drawing which is usually done under medium or high power objective of the
microscope. Every single type of cell is drawn accurately in structure, position and
proportion to each other. A sector of cells can be drawn to represent the whole structure of
the specimen if students have time constraint. In the case of plant cells, double lines can be
used to show the thickness of the cell wall. Cells should not be drawn overlapping.
Plan drawing of dicot root (t.s.) Detailed drawing of a sector of dicot root
Cells drawn overlapping (X)
8. Drawings may be done on specimens sectioned transversely, vertically or obliquely, in which
case the shape of a particular cell should be done accordingly.
Aspecimen is sectioned transversely or horizontally Aspecimen is sectioned vertically
9. Drawings of specimen should be drawn as seen with the naked eyes or as observed under a hand lens
or examined under the microscope. No extra details should be drawn out of students own
imagination.
10. Orientation of the drawing should be done according to the position of the specimen. The
specimen may be viewed from the anterior, posterior, dorsal, ventral or lateral position. When a
specimen is seen from the dorsal view, the left and right position of the student corresponds to
that of the specimen. When a specimen is placed on its dorsal side and the ventral view of the
specimen is observed; then the left and right position of the student is opposite to that of the
specimen (see following diagram).
(v.s) (t.s)
Dissection of a rat
Abasic dissecting set
B. Graph
1. Graphs should be drawn to occupy almost the full page of the graph paper.
2. A title must be given to the graph drawn. Usually, the title is written above the graph.
3. Appropriate scales should be used and the units stated.
4. Label the axes.
5. All points should be accurately plotted according to the tabulated results.
6. Draw the best line graph to pass through as many points as possible.
7. All experimental results should be tabulated.
For example, the results of a photosynthetic experiment can be tabulated as follows:
Light intensity
No. of oxygen bubbles produced
(arbitrary units)
First attempt Second attempt
A graph showing the correlation between the
oxygen produced and light intensity in Hydrilla sp.
scalpels scissors forceps magnifying
glass
DETERMINATION OF THE OSMOTIC POTENTIAL OF THE POTATO CELL SAP
This experiment enables the students to :
(a) prepare sucrose solutions of various molarities from a stock solution
(b) tabulate experimental results and draw relevant graphs
(c) analyse and interpret experimental results for osmotic potential determination
Five solutions of different concentrations were prepared from a given 1.0 M (1.0 mol dm
3
) sucrose
solution as follows :
Molarity 0.1 M 0.2 M 0.3 M 0.4 M 0.5 M
Volume of 1.0 M sucrose solution (cm
3
) 2 4 6 8 10
Volume of distilled water (cm
3
) 18 16 14 12 10
15 strips of potato tissues measuring about 46 cm in length and with a cross-section of 0.5 cm 0.5 cm
were cut. The average length of the potato strips were recorded. 3 potato strips each were placed into 5
different boiling tubes containing different molarities of sucrose solution. The initial level of the sucrose
solution was recorded for each tube. After 30 minutes, the final level of the sucrose solution and the final
length of the strips were recorded. The physical condition of the potato strips was also recorded.
Molarity
Length of potato strip (cm) Level of sucrose solution (cm)
(mol/dm
3
)
Before After Change in length Initial Final
0.1 4.0 4.2 4.2 4.1 0.20 5.5 5.6
0.2 4.0 4.1 4.1 4.1 0.10 5.5 5.8
0.3 4.0 3.9 3.8 3.8 0.15 5.5 5.7
0.4 4.0 3.8 3.8 3.9 0.20 5.5 6.0
0.5 4.0 3.7 3.7 3.6 0.30 5.5 6.0
Two graphs were drawn,
(i) a standard graph showing the relationship between the osmotic potential and the molarity of the
sucrose solution to determine the osmotic potential of the potato cell sap
(ii) a graph of the average change in the length against the molarity of the sucrose solution
Molarity 0.05 0.10 0.15 0.20 0.25 0.30 0.35 0.40 0.45 0.50 0.55
Osmotic
1.3 2.6 4.0 5.3 6.7 8.1 9.6 11.1 12.6 14.3 16.0
potential (atm)
From the two graphs, the following can be concluded.
(i) The osmotic concentration of the potato tissues in mol dm
3
of sucrose solution is 0.23 M.
At 0.23 M, the average change in length of the potato strips is zero which means that there is no
net movement of water in and out of the potato cells. The osmotic concentration of the potato
tissues is said to be isotonic to the surrounding sucrose solution.
(ii) The osmotic potential in atm. is 6.0 atm.
Using the value of osmotic concentration of the potato tissue (from (i)), the osmotic potential
can also be determined using the standard graph of osmotic potential against molarity of sucrose
solution.
Graph of the average change in length against the molarity of sucrose solution
* (This is only part of the graph to represent the shape of the actual graph obtained)
A standard graph of osmotic potential against molarity of sucrose solution
USE OF MICROSCOPE TO DETERMINE THE MAGNIFICATION AND THE ACTUAL
SIZE MEASUREMENT OF A CELL
This experiment enables students to :
(a) estimate the magnification of a drawing made under a microscope
(b) estimate the actual size of some microorganisms
(c) determine the size of a plant (onion scale) cell
A. Slides of various types of microorganisms are examined under a microscope at high power. The
image size (apparent object size) of the specimen can be estimated by placing the thumb and the
index finger on a ruler put beside the microscope (see picture below). A drawing is made and the
magnification of the drawing can be determined by using the formula:
The actual size of each microorganism can be determined using the formula :
Estimating the apparent size of a specimen
One eye looking into the
eyepiece and another
looking at the ruler placed
beside the microscope
Estimating the apparent
size of the observed
specimen as seen under
an objective power
Placing your two
fingers against the
ruler to estimate
the apparent size
Apparent size of object
Actual size of object =
Magnification of eyepiece Magnification of the objective lens used
Magnification of eyepiece Magnification of the
objective lens used Size of drawing
Magnification of the drawing =
Apparent size of object
The dimension on each specimen indicates the apparent size of the specimen as seen under high power
Eye piece magnification = 10 x
Objective lens magnification = 40 x
Microorganism Euglena Amoeba Hydra
Apparent object size 3 cm 2 cm 4 cm
Size of drawing 6 cm 4 cm 8 cm
Magnification of the drawing 10 40 6/3 = 800 x 10 x 40 4/2 = 800 x 10 40 8/4 = 800 x
Actual size 3/(10 40 ) = 75 m 2/(10 40 ) = 50 m 4/(10 40 ) = 100 m
B. To determine the size of a plant (onion scale) cell
To determine the diameter of a microscopes field of view using low power
Euglena Amoeba Hydra
3 cm
(apparent
size)
Reading Diameter of low power field of view
Orientation mm m
Horizontally 3.2 3200
Vertically 3.2 3200
Diameter of low power field of view = 3200 m
Magnification of high power objective lens = 40 x
1
The diameter of a high power field of view is of the diameter of a low power field of view =
4
3200
= 800 m
4
Number of cells length-wise Number of cells width-wise
First count 3 7
Second count 4 8
Third count 3 7
Average 3 7
* The figures given in the above table are meant as examples and should not be taken as true information.
Diameter of microscopes
field of view (high power) 800
Average length of one onion scale cell = = = 267 m
Number of cells length-wise 3
Diameter of microscopes field of view 800
Average width of one onion scale cell = = = 114 m
Number of cells width-wise 7
OBSERVATION OF PLANT AND ANIMAL CELLS
This experiment enables students to:
(a) prepare slides of animal (cheek) cells and plant (leaf epidermal) cells using the correct staining
technique
(b) realise that cell is a basic unit of life
A. Observation of animal cells
A toothpick is used to gently scrape off a thin layer of cells from the inside of your cheek.
The scraping is mounted in a drop of methylene blue solution on a slide and examined under low
power objective lens followed by high power objective lens.
The cells that line the inner cheek are squamous cells and their nucleus are stained purplish blue using methylene blue solution
B. Observation of plant cells
The epidermal layer of the onion scale leaf is peeled off using a pair of forceps and mounted in a
drop of iodine on a slide. The specimen is then examined under a microscope.
The onion scale leaf cells do not contain chloroplasts because the cells are storage cells for carbohydrates
The differences between animal and plant cells are :
(a) animal cells possess only cell membrane whereas plant cells possess cell membrane and cell
wall
(b) the nucleus of the animal cell is centrally positioned whereas the nucleus of the plant cell is
pushed to the periphery
Magnification = 700 x
Magnification = 900 x
ENZYME ACTIVITY
(a) investigate the effect of temperature on enzyme-catalysed reactions
(b) determine the optimal temperature of enzymatic reactions
(c) determine the temperature coefficient, Q
10
, of an enzyme-controlled reaction
A saliva solution is prepared by spitting saliva into a clean beaker and diluting it with an equal amount
of distilled water. Five beakers of water baths at the following temperatures: 0 C, 20 C, 37 C, 50C,
and 65 C, are prepared and labelled A to E. A test tube containing a mixture from the test tube is taken
out every minute and tested with iodine on a white tile. The time taken for each complete
1
hydrolysis is recorded and a graph of the reaction rate () against the temperature is plotted.
t
1
Test tube Temperature (C) Time taken for complete hydrolysis, t (minute) Rate of reaction ()
t
A 0 105 0.01
B 20 35 0.03
C 37 10 0.10
D 50 100 0.01
E 65 00 0.00
1
Graph of reaction rate () against temperature
t
From the graph, the following can be inferred.
(a) The temperature coefficient, Q
10
, between 30 C and 40 C
rate of reaction at 40 C 0.094
= = = 1.709
rate of reaction at 30 C 0.055
(b) The value of Q
10
is about 2, that is, the rate of reaction which is controlled by enzyme, doubles for
every 10 C increase in temperature provided that the temperature does not exceed optimum
temperature.
(c) At low temperatures, the rate of enzyme-catalysed biochemical reaction increases with temperature in
such a way that the rate doubles for every 10 C increase in temperature until the rate reaches the
maximum at an optimum temperature of about 37 C. Beyond this temperature, the rate decreases
with an increase in temperature as some of the enzymes become denaturated and the reaction stops at
65 C.
SEPARATION OF PHOTOSYNTHETIC PIGMENTS USING PAPER
CHROMATOGRAPHY
(a) prepare a concentrated chlorophyll extract from pandan leaves
(b) separate the pigments found in the chlorophyll extract
(c) calculate the R
f
value of each separated pigment
The pigment extract is prepared by blending some pandan leaves with acetone and filtering it
through a muslin cloth. A chromatography strip is cut and a pencil baseline is drawn on the strip. The
pigment extract is then transferred onto the chromatography strip using a dropper to make a
concentrated spot in the middle of the baseline. The strip is then placed in a boiling tube containing
petroleum-ether solvent. After 30 minutes, the solvent and pigment fronts are marked and their
distances from the base are measured.
Chromatography paper
Main pigment colour Chlorophyll b Chlorophyll a Xanthophyll Carotene
(green) (blue green) (yellow) (yellow orange)
Distance moved by pigment (mm) 32 64 73 95
Distance moved by solvent (mm) 97 97 97 97
Distance moved by pigment
R
f
= 0.33 0.66 0.75 0.98
Distance moved by solvent
In paper chromatography, different pigments are carried up the chromatography strip at different
speeds due to the following factors.
(i) Adsorption ability of the paper to the solutes to be separated caused by the porosity of the paper
(ii) Solubility of the solutes in a particular solvent
(iii) The molecular mass of the solute; the densest will be the last to move up the paper
Pandan leaves contain four distinct pigments with their R
f
values as shown in the table above.
solvent front
xanthophyll (yellow)
chlorophyll a (blue-green)
chlorophyll b (green)
spot of concentrated
chlorophyll extract
baseline
carotene (yellow-orange)
phaeophytin (grey)
EXAMINATION OF SLIDES - TRANSVERSE SECTIONS OF THE C
3
AND C
4
PLANT
LEAVES
This experiment enables students to :
(a) relate the structure of the leaf cell to its functions
(b) differentiate the anatomical leaf structure of C
3
and C
4
plants in relation to the Hatch-Slack
pathway and Calvin cycle
Slides of transverse sections of the C
3
and C
4
leaves are examined under the microscope at low power to
observe the plan view of the leaf sections. High power labelled drawing for the cells observed are then
made.
High power drawing of cross section of C
3
leaf
In the C
3
plant leaf, the palisade mesophyll cells exist as one basic layer. Its bundle sheath cells are
small and the intercellular spaces are bigger than in the leaf of a C
4
plant. The carbon dioxide
receptor in the Calvin cycle is ribulose bisphosphate.
Magnification of drawing 4 600 = 2400 x
14
Magnification of drawing 600 = 2800 x
3
High power drawing of cross section of C
4
leaf
In the C
4
plant leaf, the mesophyll cells are arranged to form a ring surrounding the bundle sheath
cells. The bundle sheath cells are large and contain chloroplasts. The C
4
pathway first uses
phosphoenolpyruvate to fix carbon dioxide to form oxaloacetate in the mesophyll cells which later
release the gas to the bundle sheath cells where the carbon dioxide receptor is ribulose bisphosphate
(Calvin cycle).
The outer layer of the leaf is a single-celled thick epidermis lined with a thin layer of waxy cuticle
which prevents excessive loss of water through transpiration. This layer protects the leaf from
pathogens and it is transparent to allow sunlight to pass through.
DISSECTION OF THE MAMMALIAN (RAT) DIGESTIVE SYSTEM
This experiment enables students to display:
(a) the digestive system and the related organs
(b) the blood vessels, arteries and veins of the system
The rat is pinned to a dissecting board, with the ventral side facing upwards. Its abdominal cavity is
cut to display the viscera organs to the left side of the animal. Three labelled drawings are made
showing the alimentary canal, related organs, and their veins and arteries.
Magnification of drawing = 2 x
xiphoid cartilage
liver
stomach
intestine
Alimentary canal and related organs on the left side of rat
Alimentary canal and related organs on the right side of rat
The hepatocytes in the liver produce bile which is secreted into the duodenum which helps to
emulsify fats to facilitate the lipase enzyme in digesting it.
The pancreas produces and secretes pancreatic juices which contain digestive enzymes such as
amylase, trypsin and lipase to hydrolyse food substances.
DISSECTION OF THE MAMMALIAN (RAT) RESPIRATORY SYSTEM
(a) examine the structures of the main organs involved in respiration
(b) increase their understanding of the process of gaseous exchange in animals
The rat is pinned to the dissecting board with the ventral side upwards. An incision is made through
the skin and is cut as far as the lower jaw. The ventral and lateral thoracic walls are then cut along the
dotted lines to expose the thoracic cavity (see diagram below). The muscles and tissues of the neck
are also cut to expose the trachea and larynx. The heart, lungs, trachea, oesophagus, and larynx are
removed together. Labelled drawings of the structures taken out are made.
There are 7 pairs of ribs found in the rat. When the ribcage is raised upwards and forwards, the volume of
the ribcage increases, lowering the pressure inside. Air is forced into the lungs through the nostrils,
trachea and other respiratory tubes. This is inhalation. During exhalation, the ribcage falls downwards
and inwards due to gravity, decreasing its volume but increasing its pressure. Air is then forced out.
The diaphragm is a membranous structure with peripheral elastic muscles radiating from the centre
forming a dome covering the base portion of the airtight ribcage. During inspiration, the radial
muscle of the diaphragm contracts, flattening its dome-shaped structure. This causes the volume of
the ribcage to increase. During expiration, the radial muscle relaxes, returning the diaphragm to its
dome-shaped structure and the volume of the ribcage returns to its smaller capacity.
The left lung consists of a single lobe whereas the right lung has 4 lobes. When examined under a
hand lens, a cut lung tissue looks soft and spongy. There are many thin structures with air spaces and
blood capillaries in it.
The length of the trachea when measured from the larynx to the point where it branches into two
bronchi is about 2.5 cm.
DISSECTION OF THE MAMMALIAN (RAT) CIRCULATORY SYSTEM
(a) identify the organs in the thoracic cavity
(b) identify the position of the main veins and arteries and their branches
A rat is pinned to the dissecting board with the ventral side upwards. A mid-ventral incision through
the skin is made and cut towards the mouth and then towards the posterior. The xiphoid cartilage is
pulled downward and the diaphragm is cut. The ventral and lateral thoracic walls are then cut (see
diagram below- cut along the line) to expose the thoracic cavity. The muscles and tissues of the neck
are also cut to expose the trachea and larynx. Labelled drawings of the veins and arteries in the
thoracic region of the rat are made.
Dorsal view of the heart (photo)
xiphoid cartilage
stemum
ribcage
lungs
liver
The veins in the thoracic region of rat
Artery on both sides of the thorax
Dorsal view of the heart
carotid artery
aortic arch
left anterior vena cava
right anterior vena cava
ductus arteriosus
pulmonary vein
entering left auricle
left auricle
vena cava
entering
right auricle
left ventricle
right ventricle
posterior vena cava
right auricle
pulmonary artery
subclavian artery
EXAMINATION OF PREPARED SLIDES OF LIVER AND KIDNEY
(a) understand the structures of the liver and kidney
(b) understand the functions of liver and kidney as homeostatic organs
The prepared slides of the tranverse sections of the liver and kidney were observed under microscope
at low power to determine the distribution of all the tissues. The detailed structures were then seen
under high power. Plan and detailed drawings of the slides were then made.
Plan drawing of slide of liver- transverse section
Plan drawing of slide of kidney- transverse section
cortex
medulla
nephron
pelvis
renal artery
bile duct
intralobular
vein - a
tributary of
hepatic vein
lobule of liver
branch of
hepatic artery
branch of
hepatic portal
vein
Magnification of drawing : 250 x
Detailed drawing of slide of liver- transverse section
Detailed drawing of slide of kidney-transverse section
The liver is fed with three types of blood vessels namely the hepatic artery, the hepatic vein and the
hepatic portal vein. The liver performs many functions; one of which is the production of bile. The
bile produced contains many substances and salts, sodium glycocholate and sodium taurocholate
which can emulsify fats, that is, the salts are able to break the fats or oil into smaller droplets and
separate them permanently to facilitate digestion. These salts are not enzymes because they do not
break down the fats chemically. After digestion, simple food substances are absorbed. After
assimilation and metabolism, excretary products (by-products of cell metabolism) especially,
ammonia needs to be removed from the body as it is toxic and changes the pH of the internal
environment of the body.
basal
membrane
of capsule
glomerulus
distal
convoluted
tubule
nucleus of
podocyte
lumen of
Bowmans
capsule
proximal
convoluted
tubule
microvilus
Bowmans
capsule
sinusoid
hepatocytes
Kupffer cell

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