5.1Theme A — Unity and diversity
Tool 1: Experimental Techniques — IB Biology SL Revision Notes & Topical Questions
10 min read 13 parts Lesson 2 of 11 Unit quiz
Lesson summary
5.1 Tool 1: Experimental Techniques — IB BIOLOGY SL notes: Select and use suitable apparatus and techniques to make measurements with stated uncertainty, and keep practical work safe, ethical and fair.Hazard = a feature of a procedure that has inherently harmful propertiesA chemi…
- Risk assessment
- Justified precautions
- Hazard warning symbols
- Human participants
- Field work
- Range and repeats
Part 1 of 11Hazard, risk and risk assessment
Hazard
Hazard a feature of a procedure that has inherently harmful properties
- A chemical may be an irritant to the skin.
- A rocky shore may have slippery rocks.
- A Bunsen burner may cause burns.
Risk
Risk the level of danger posed by a hazard, decided by how likely the harm is and how severe it would be
Risk assessment
- Risk assessment identifies the hazards, assesses the risk and changes the method to reduce it
- Rocky shore exampleslippery rocks are a hazard and slipping is quite likely.
- The procedure is altered so that footwear has good grip, students avoid seaweed-covered rocks and walk slowly.
Justified precautions
- Justified precautions name the hazard and explain why the precaution reduces the risk
- Exampleiodine is an irritant, so wear safety goggles and wash any skin it touches at once.
Part 2 of 11Hazard symbols, ethics and environment
Hazard symbols, ethics and environment
Hazard warning symbols
- Hazard warning symbols nine standard symbols, usually in a red-bordered diamond on labels and safety data sheets
- Examples are health hazard, corrosive, flammable, acute toxicity, oxidising and hazardous to the environment.
- Check them for stains, enzyme reagents and chromatography solvents.
Animals
- IB policy says to use animals only if no alternative exists
- Investigations must not be cruel and must remove causes of distress.
- Keep experiments short, handle animals as little as possible and release them to their habitat afterwards.
Human participants
- Human participants any investigation, such as measuring breathing with a spirometer, needs fully informed consent
Laboratory waste
- No toxic chemicals down the sink.
- Sharps go into a sharps bin.
- Biological waste, such as cultures or dissected organs, is sterilised or disposed of correctly.
Field work
- Field work work in small groups, avoid sensitive habitats and take away all equipment and litter
Part 3 of 11Designing a fair test
Fair test
- Fair test changes one independent variable, measures one dependent variable and keeps all other significant variables constant
- Name each controlled variable and the method used, such as a water bath at 30 °C.
Range and repeats
- Range and repeats use at least five values of the independent variable and at least three repeats of each
- This lets trends and anomalies be identified.
Control
Control a treatment identical to the others except that the independent variable is absent or at a baseline level
Independent variable
- Independent variable deliberately changed and plotted on the x-axis
Dependent variable
- Dependent variable measured in response and plotted on the y-axis
Part 4 of 11Uncertainty and error
- Instrument uncertainty
- Analogue scales are read to ± half the smallest division.
- Digital displays are read to ± the last digit.
- Record the uncertainty in the table heading, for example Volume / cm3 ± 0.05.
| Feature | Random error | Systematic error |
|---|---|---|
| Effect | Scatter about the true value | All values shifted one way |
| Affects | Precision | Accuracy |
| Reduced by | Repeats and a mean; a more sensitive instrument | Calibration, zeroing, improving the method |
| Example | Judging a colour end-point by eye | A balance not tared |
Accuracy
Accuracy how close a measurement is to the true value
Precision
Precision how close repeated measurements are to each other
Part 5 of 11Measuring instruments
| Quantity | Instrument | Units | Notes |
|---|---|---|---|
| Mass | Electronic balance | g, mg | Tare before use |
| Volume | Pipette, syringe, measuring cylinder, burette | cm3, ml | Read the bottom of the meniscus at eye level |
| Time | Stopwatch or data logger | s, min | Human reaction time adds about 0.2 s of random error |
| Temperature | Thermometer, probe or water bath | °C | Let solutions equilibrate before mixing |
| pH | pH meter or universal indicator | none | Calibrate the meter with buffers |
| Length | Ruler or eyepiece graticule | mm, µm | Recalibrate the graticule at each magnification |
| Concentration or colour | Colorimeter | Absorbance | Zero with a blank |
Choosing volume apparatus
- A measuring cylinder gives approximate volumes.
- A volumetric pipette gives one fixed volume, such as 25 cm3, most accurately.
- A burette gives accurate variable volumes up to 50 cm3 and reads from zero at the top.
Gases
- Gases collect in a gas syringe, or in an inverted cylinder of water if the gas is insoluble
Part 6 of 11Dilution and serial dilution

Dilution equation
- C1V1 = C2V2, where C is concentration and V is volume
Dilution factor
Dilution factor final volume divided by the volume of stock added
Serial dilution
- Serial dilution each step uses the same dilution factor and the concentrations change logarithmically
- It is useful for bacterial counts or sucrose solutions for osmosis.
1
Make a 1 mol dm-3 stock solution and label it.
2
Set out labelled tubes each holding 9.0 cm3 of distilled water.
3
Transfer 1.0 cm3 of stock to tube 1 with a clean pipette and mix; this gives 0.1 mol dm-3.
4
With a fresh pipette transfer 1.0 cm3 from tube 1 to tube 2 and mix to give 0.01 mol dm-3.
5
Repeat for further tubes to give 10-3, 10-4 mol dm-3.
6
Use each dilution in the investigation, with at least three repeats per concentration.
Part 7 of 11Microscopy and magnification
Temporary mount
- Temporary mount cut a thin section and add a drop of water or stain such as iodine or methylene blue
- Lower the coverslip at an angle to avoid air bubbles.
- Focus with low power first, then use only the fine focus on higher power.
Magnification
- M = I / A,
- so actual size A = I / MWhat the symbols mean: M = magnification (no unit) · I = image size · A = actual size; use the same unit for both (1 mm = 1000 μm)
- so actual size A = I / M
- I and A must be in the same units and M has no units.
- 1 mm = 1000 µm and 1 µm = 1000 nm.
Resolution
Resolution the minimum distance between two points that can still be seen as separate
- It is about 0.2 µm for a light microscope and 0.2 nm for an electron microscope.
1
Place the stage micrometer (1 division = 10 µm) on the stage and focus at the chosen objective.
2
Turn the eyepiece so the graticule scale lies alongside the micrometer scale.
3
Find two points where lines coincide and count the divisions on each scale between them.
4
Calculate 1 eyepiece division = (stage divisions × 10 µm) / eyepiece divisions.
5
Replace the micrometer with the specimen, measure in eyepiece divisions and multiply by the calibration value.
6
Recalibrate for every change of objective lens.
Part 8 of 11Chromatography and colorimetry
Chromatography
- Chromatography separates photosynthetic pigments by their solubility in the solvent and attraction to the paper or TLC plate
- Mark the origin in pencil and keep the spot above the solvent level.
- Mark the solvent front immediately when the paper is removed.
Rf
Rf distance moved by the pigment divided by distance moved by the solvent front
- It has no units and is always between 0 and 1.
Colorimetry
- Colorimetry measures the absorbance of light by a coloured solution
- Make a calibration curve from standards of known concentration.
- Read the unknown concentration from the curve.
- Zero the colorimeter with a blank of solvent.
Part 9 of 11Rates, counts and field sampling
Measuring rates
- Collect gas in a gas syringe or inverted measuring cylinder rather than counting bubbles.
- A respirometer with KOH or soda lime to absorb CO2 measures O2 uptake.
- A potometer measures water uptake as an estimate of transpiration.
- Rate = amount of product formed (or substrate used) divided by time.
Cell counts
- Cell counts a haemocytometer is a slide with a grid of known volume
- Counting cells in the grid allows the concentration of the whole culture to be calculated.
- Direct counts need no specialist equipment but are slow.
Random quadrats
- Random quadrats use random number coordinates to compare two areas
- Population density = mean number of individuals per quadrat divided by the area of one quadrat.
Transects
- Transects line or belt transects show change along a gradient
Mark-release-recapture
- Mark-release-recapture population size N = (M × C) / R
- M is the number first marked, C the total caught second time and R the number recaptured marked.
- It assumes no births, deaths or migration and that marks do not affect survival or get lost.
Part 10 of 11Aseptic technique and observation
Aseptic technique
- Aseptic technique procedures that prevent contamination of cultures and protect the worker
- Sterilise loops by flaming until red hot.
- Open lids as little as possible.
- Tape Petri dishes without sealing them fully.
- Incubate at 25 °C, not 37 °C, to avoid culturing human pathogens.
- Dispose of cultures by autoclaving.
Biological drawings
- Biological drawings labelled line drawings show the specific features seen in the specimen
Qualitative data
- Qualitative data describe attributes that are observed but not measured, such as fur colour, sex or number of limbs
- They are used when identifying and classifying organisms.
Required techniques
- Required techniques chromatography, colorimetry and serial dilution, modelling, light microscopy, temporary mounts, sampling, karyotyping and cladogram analysis
Part 11 of 11More from the specification
Safety in biology
- Safety in biology risk assessment identifies hazards such as sharp tools, microorganisms and chemicals and the steps to reduce them
Ethical considerations
- Ethical considerations involve respect for living organisms, minimising harm and justifying any animal or human use
Environmental issues
- Environmental issues disposal of waste and effects on habitats must be considered when making observations in the field
Applying techniques
- Applying techniques experimental techniques involve careful measurement, controls and repeats, for example for a liquid culture or sampling
Key terms
| Term | Meaning |
|---|---|
| Accuracy | How close a measured value is to the true value. |
| Precision | How close repeated measurements are to each other. |
| Controlled variable | A factor other than the independent variable that is kept constant. |
| Serial dilution | A stepwise series of dilutions that each use the same dilution factor. |
| Resolution | The minimum distance between two points at which they can still be seen as separate. |
| Retention factor (Rf) | Distance moved by the pigment divided by distance moved by the solvent front. |
| Quadrat | A square frame of known area used to sample organisms and estimate density. |
| Aseptic technique | Procedures that prevent unwanted microorganisms contaminating cultures. |
Avoid these mistakes
1
Confusing hazard and risk — a hazard is something that can cause harm and risk is the likelihood and severity of that harm
2
Stating only wear a lab coat and goggles as a precaution — name the hazard and justify the precaution
3
Mixing units in M = I / A — convert I and A to the same units first
4
Thinking repeats remove systematic error — repeats reduce random error and only calibration removes systematic error
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