MICROSCOPY

The stage micrometer is 1 mm long with 100 divisions so each division of the stage micrometer is
one one-hundredth of a mm (0.01mm or 10 um).

1x106 nm= 1x103 µm = 1mm = 0.1 cm

objective Magnification Number of ocular Number of stage Dimension of one

divisions micrometer divisions ocular division (one
(eyepiece eyepiece graticule
graticule division)
divisions)

scanning 4x 40 10 25 40 ocular divisions = 10 stage

micrometer divisions.
Thus 40 ocular divisions = 100um
1 ocular division = 25um

Low power 10x 100 10 10 100 ocular divisions = 10 stage

micrometer divisions
Thus 100 ocular divisions = 100 um
1 ocular division = 2.5um

High power 40x 40 1 2.5 40 ocular divisions = 1 stage micrometer

divisions.
Thus 40 ocular divisions = 10um
1 ocular division = 2.5um
DRAWINGS
Plan diagrams (10x objective lens) High power drawings (40x objective lens) General biological drawings
(not examined under a
microscope

The purpose of a low power drawing is usually to The purpose of high power drawings is to show as
show the distribution of the main tissues within an much accurate detail as microscopy will allow
organ, for example in a transverse section of a stem
or a trachea. Draw only a few representative adjacent cells.

Draw only the outline of the tissue. If all the cells are similar, then three cells is often
sufficient to show both cell structure and the way in
Students are required only to identify the tissues and which cells are arranged in relation to each other.
to delimit the different tissues with boundary lines.

C clean continuous lines

S scale (70% of the whole space allocated)

P proportion of drawings to different elements

D details (draw only what you see and include everything you see)

L Labels (label only if instructions say so)

 DILUTION
Simple dilution Serial dilution

- Don’t take from the test tube sample - Taken from the previous dilution sample
- Only reagents needed are water and the solution - Same volume of solution is used from the
added in different amounts previous sample when added to the next test
tube
- Concentration is half of the previous sample

FOOD TESTS
test test + results procedure

Iodine test ● blue black = starch present ● Add iodine solution to sample dropwise
● remains yellowish brown = starch
absent

Benedict’s test for reducing ● brick red/ orange precipitate = ● Add equal volumes of benedict’s
sugars presence of reducing sugars solution to equal volume of food
● yellow-orange/ green suspension samples (mixed with equal volumes
= minimal reducing sugars distilled water) in a test tube
present ● Place test tubes in boiling water and
● solution remains blue = no observe for colour changes
reducing sugars
*Sucrose is a non reducing sugar

Non reducing sugars test ● brick red/ orange precipitate = ● Add equal volumes of HCL to samples
presence of nonreducing sugars in test tube and boil for 1 minute
● yellow-orange/ green suspension ● Let it cool and add NAHCO3 and wait
= minimal non reducing sugars for effervescence to stop
present ● Repeat benedict’s test
● solution remains blue = no non
reducing sugars *HCL → boil → NAHCO3

Biuret test ● turns violet = presence of ● Add biuret solution and sample into test
proteins tube in equal amounts dropwise
● remains pale blue = no proteins ● Preparation of biuret reagent: add
NaOH to sample adn shake. One
minute later add CuSO4 dropwise.
Ethanol emulsion test ● white emulsion = presence of ● Add equal volumes of ethanol and
lipids sample into test tube
● no emulsion = no lipids ● Shake vigorously
● Add equal volume of distilled water to
the solution and mix well

SPECTROPHOTOMETER
● determines how much light of a particular colour is absorbed by a liquid sample.
● more coloured substance in the solution = more light will be absorbed
(i.e. the less light passes through the solution).
calculates the amount of light absorbed by the sample.

*remember to put the clear sides sideways so that light can pass through. Mate side faces forward

GEL ELECTROPHORESIS

Purpose of loading dye Purpose of TAE buffer

● Glycerol helps DNA sample to sink into ● It contains ions that will conduct electricity;
the wells ● It contains a pH buffer (pH 8.3) to ensure
● Allows the DNA to be coloured to track that DNA has a net negative charge and
migration of fragments\ migrates towards the positive electrode

TABLES AND GRAPHS

Heading Appropriate column / row headings.

Precision Raw data recorded and consistency in decimal

Trend Correct trend observed

Complete All data are recorded

Graph type ● Line graph, bar graph, histogram
○ Dot-to-dot plot is appropriate when data may show no clear
relationship
○ Best fit line is appropriate where data shows an underlying
relationship or a trend is required

Axis ● Correct labels along with choice of axis

*for bar graphs there is no x axis just label every individual column

Points ● Points plotted correctly

● Use small crosses

Scale ● Draw to occupy at least 70% of the space given

WORKING
● List everything step by step
● Write out statements

SOURCES OF ERROR
errors solutions

1. Pipette/ syringe accuracy Use a micropipette for increased precision

2. Cutting accuracy Use a microtome

3. Not from the same parent Ensure that all samples are from the same origin

4. temperature Use thermostat controlled water bath

5. Colour perception Use spectrophotometer

6. Volume Use burette

7. Unreliable results Repeat experiment

TOPICS TESTED FOR PRACTICAL
Cell Structure
● Use of microscope for observation of Nerium leaf*
● Wet mount and staining of onion epidermis*

Cell Membrane
● Effect of pH of cell membrane permeability
● Effect of different solvents on cell membrane permeability*
● Effect of temperature on cell membrane permeability

Biomolecules (Carbohydrates, Lipids, Proteins)

● Identification of biomolecules present in specimens*
○ Conducting of food tests
● Quantify concentration of biomolecules present in unknown solution*

Enzymes
● Effect of amylase concentration on digestion of starch*
● Effect of surface area to volume ratio of potato strips on H2O2 decomposition*
○ The potato strips contain the enzyme for H2O2 decomposition
○ Increase in surface area = more EX complex formation = increase in rate of reaction
● Effect of H2O2 concentration on its decomposition by yeast-alginate beads*
○ H2O2 is the substrate
○ So an increase in substrate concentration = increase the rate of ES complex formation.
● Effect of temperature on enzyme activity
● Effect of pH on enzyme activity
● Effect of concentration of inhibitors on enzyme activity

Cell Cycle
● Use of microscope for observation of onion root tip (mitosis)*
● Use of microscope for observation of anther (meiosis)

Molecular techniques
● Paternity testing using gel electrophoresis*
● Determine phylogenetic relationship between plants*

Infectious diseases
● Use of microscope for observation of human blood sample*
● Blood typing*
● Effect of concentration of penicillin on bacterial growth*

Photosynthesis
● Effect of light intensity on rate of photosynthesis using DCPIP*
○ DCPIP an electron acceptor that is blue when oxidised and colourless when reduced.
○ Hence when the electron transport chain proceeds, it accepts the electron and become
colourless as it is reduced
○ So increased rate of photosynthesis = increased rate of blue turning colourless
● Effect of wavelengths of light on rate of photosynthesis using DCPIP
● Effect of bicarbonate concentration on rate of photosynthesis of hydrilla (water plant)
Respiration
● Effect of concentration of glucose on rate of yeast respiration*
○ Increase in glucose = increase rate of respiration
● Effect of types of carbohydrates on rate of yeast respiration*
○ Glucose is allows for the greatest rate of respiration as it is the raw material required
● Effect of temperature on rate of respiration of mealworms