Investigating Mitosis

Preparation of Stained Squashes of Cells from Root Tips

Only a few cells are able to continue dividing in a multicellular organism. In plants, the growing tips of roots and shoots contain meristem tissue that can divide by mitosis for growth.

1) Sample preparation

Wear gloves and use forceps to handle the tips.
Root tips must be sprouting (actively growing).
Place into 5 M hydrochloric acid.
After 5 minutes, rinse the tips in cold water in a watch glass.

2) Cut the root tips

Using a sharp scalpel, cut root tips that are 2 mm long.
Place a root tip onto a microscope slide. Ensure the slide is clean to reduce the chances of artefacts.

3) Staining

Carefully add 2-3 drops of stain and leave for two minutes.
Use a mounted needle to spread out the root tips into a thin layer.
Place a coverslip over the top of the tips.

4) Squashing

Squash down by applying force to the cover slip. This could be with the flat end of a pencil, or the slide could be covered with a paper towel and pressed.
Force must be vertical or the cover slip may break and cause injury.

5) Viewing the sample

Place the slide on the microscope stage using the lowest power lens.
Focus the lens on the sample using first the coarse control and then the fine control.
Move the slide to see the range of cells. The cells closer to the tip will be those more actively dividing.
On a lens power of 400x, it should be possible to clearly see the chromosomes in the dividing cells.

Mitotic Index

The mitotic index is used to work out the proportion of cells in the sample that are in mitosis.

Equation

The equation for mitotic index for a particular field of view is:
- Mitotic index = number of cells in mitosis ÷ total number of cells

E.g. Mitotic index

In a field of view, 29 cells are counted in total.
22 of the 29 cells have visible chromosomes and are actively dividing.
- 22 ÷ 29 = 0.7586
- About 76% of the cells in the sample are undergoing mitosis.

Cell Size

The size of a cell can be calculated by using a micrometer and graticule.

Calculating cell size

The eyepiece graticule is a glass disc in the eyepiece of a microscope.
Cells can be seen through a microscope and a micrometer can be used to calculate the actual size of the cells.

Calibrating the eyepiece

In order to calculate cell size, the eyepiece graticule must be calibrated.
The graticule is calibrated by comparing the length of each division in the micrometer to the length of each division in the graticule.

E.g. Graticule calibration

E.g. 100 divisions on the eyepiece graticule are equal to 25.9 divisions on the micrometer.
- In the micrometer, 100 divisions = 1mm.
- This means that 1 division = 10µm and 100 eyepiece divisions = 25.9 × 10µm = 259µm.
- This means that 1 eye piece division = 259 ÷s; 100µm = 2.59µm.
- The ratio of micrometer division to eyepiece division is 10µm to 2.59µm.

Calculating actual cell size

Once the eyepiece graticule has been calibrated, actual cell size can be calculated using the equation:
- Actual size = size of image ÷ magnification

E.g. Actual cell size

The length of a cell is measured to be 33mm in an image drawn at a magnification of 400x.
- Size of image = 33mm = 33000 µm.
- Magnification = 400x.
- Actual size = 33000 ÷ 400 = 82.5µm.

1Biological Molecules

1.1Monomers & Polymers

1.1.1Monomers & Polymers

1.1.2Condensation & Hydrolysis Reactions

1.2Carbohydrates

1.2.1Structure of Carbohydrates

1.2.2Types of Polysaccharides

1.2.3End of Topic Test - Monomers, Polymers and Carbs

1.2.4Exam-Style Question - Carbohydrates

1.2.5A-A* (AO3/4) - Carbohydrates

1.3Lipids

1.3.1Triglycerides & Phospholipids

1.3.2Types of Fatty Acids

1.3.3Testing for Lipids

1.3.4Exam-Style Question - Fats

1.3.5A-A* (AO3/4) - Lipids

1.4Proteins

1.4.1The Peptide Chain

1.4.2Investigating Proteins

1.4.3Primary & Secondary Protein Structure

1.4.4Tertiary & Quaternary Protein Structure

1.4.5Enzymes

1.4.6Factors Affecting Enzyme Activity

1.4.7Enzyme-Controlled Reactions

1.4.8End of Topic Test - Lipids & Proteins

1.4.9A-A* (AO3/4) - Enzymes

1.4.10A-A* (AO3/4) - Proteins

1.5Nucleic Acids

1.5.1DNA & RNA

1.5.2Nucleotides

1.5.3Polynucleotides

1.5.4DNA Replication

1.5.5Exam-Style Question - Nucleic Acids

1.5.6A-A* (AO3/4) - Nucleic Acids

1.6ATP

1.6.1Structure of ATP

1.6.2Hydrolysis of ATP

1.6.3Resynthesis of ATP

1.6.4End of Topic Test - Nucleic Acids & ATP

1.7Water

1.7.1Importance of Water

1.7.2Structure of Water

1.7.3Properties of Water

1.7.4A-A* (AO3/4) - Water

1.8Inorganic Ions

1.8.1Inorganic Ions

1.8.2End of Topic Test - Water & Inorganic Ions

2Cells

2.1Cell Structure

2.1.1Introduction to Cells

2.1.2Eukaryotic Cells & Organelles

2.1.3Eukaryotic Cells & Organelles 2

2.1.4Prokaryotes

2.1.5A-A* (AO3/4) - Organelles

2.1.6Methods of Studying Cells

2.1.7Microscopes

2.1.8End of Topic Test - Cell Structure

2.1.9Exam-Style Question - Cells

2.1.10A-A* (AO3/4) - Cells

2.2Mitosis & Cancer

2.2.1Mitosis

2.2.2Stages of Mitosis

2.2.3Investigating Mitosis

2.2.4Cancer

2.2.5A-A* (AO3/4) - The Cell Cycle

2.3Transport Across Cell Membrane

2.3.1Cell Membrane Structure

2.3.2A-A* (AO3/4) - Membrane Structure

2.3.3Diffusion

2.3.4Osmosis

2.3.5Active Transport

2.3.6End of Topic Test - Mitosis, Cancer & Transport

2.3.7Exam-Style Question - Membranes

2.3.8A-A* (AO3/4) - Membranes & Transport

2.3.9A-A*- Mitosis, Cancer & Transport

2.4Cell Recognition & the Immune System

2.4.1Immune System

2.4.2Phagocytosis

2.4.3T Lymphocytes

2.4.4B Lymphocytes

2.4.5Antibodies

2.4.6Primary & Secondary Response

2.4.7Vaccines

2.4.8HIV

2.4.9Ethical Issues

2.4.10End of Topic Test - Immune System

2.4.11Exam-Style Question - Immune System

2.4.12A-A* (AO3/4) - Immune System

3Substance Exchange

3.1Surface Area to Volume Ratio

3.1.1Size & Surface Area

3.1.2A-A* (AO3/4) - Cell Size

3.2Gas Exchange

3.2.1Single-Celled Organisms

3.2.2Multicellular Organisms

3.2.3Control of Water Loss

3.2.4Human Gas Exchange

3.2.5Ventilation

3.2.6Dissection

3.2.7Measuring Gas Exchange

3.2.8Lung Disease

3.2.9Lung Disease Data

3.2.10End of Topic Test - Gas Exchange

3.2.11A-A* (AO3/4) - Gas Exchange

3.3Digestion & Absorption

3.3.1Overview of Digestion

3.3.2Digestion in Mammals

3.3.3Absorption

3.3.4End of Topic Test - Substance Exchange & Digestion

3.3.5A-A* (AO3/4) - Substance Ex & Digestion

3.4Mass Transport

3.4.1Haemoglobin

3.4.2Oxygen Transport

3.4.3The Circulatory System

3.4.4The Heart

3.4.5Blood Vessels

3.4.6Cardiovascular Disease

3.4.7Heart Dissection

3.4.8Xylem

3.4.9Phloem

3.4.10Investigating Plant Transport

3.4.11End of Topic Test - Mass Transport

3.4.12A-A* (AO3/4) - Mass Transport

4Genetic Information & Variation

4.1DNA, Genes & Chromosomes

4.1.1DNA

4.1.2Genes

4.1.3Non-Coding Genes

4.1.4The Genetic Code

4.1.5A-A* (AO3/4) - DNA

4.2DNA & Protein Synthesis

4.2.1Protein Synthesis

4.2.2Transcription & Translation

4.2.3End of Topic Test - DNA, Genes & Protein Synthesis

4.2.4Exam-Style Question - Protein Synthesis

4.2.5A-A* (AO3/4) - Coronavirus Translation

4.2.6A-A* (AO3/4) - Transcription

4.2.7A-A* (AO3/4) - Translation

4.3Mutations & Meiosis

4.3.1Mutations

4.3.2Meiosis

4.3.3A-A* (AO3/4) - Meiosis

4.3.4Meiosis vs Mitosis

4.3.5End of Topic Test - Mutations, Meiosis

4.3.6A-A* (AO3/4) - DNA,Genes, CellDiv & ProtSynth

4.4Genetic Diversity & Adaptation

4.4.1Genetic Diversity

4.4.2Natural Selection

4.4.3A-A* (AO3/4) - Natural Selection

4.4.4Adaptations

4.4.5Investigating Natural Selection

4.4.6End of Topic Test - Genetic Diversity & Adaptation

4.4.7A-A* (AO3/4) - Genetic Diversity & Adaptation

4.5Species & Taxonomy

4.5.1Courtship Behaviour

4.5.2Phylogeny

4.5.3Classification

4.5.4DNA Technology

4.5.5A-A* (AO3/4) - Species & Taxonomy

4.6Biodiversity Within a Community

4.6.1Biodiversity

4.6.2Index of diversity

4.6.3Agriculture

4.6.4End of Topic Test - Species,Taxonomy& Biodiversity

4.6.5A-A* (AO3/4) - Species,Taxon&Biodiversity

4.7Investigating Diversity

4.7.1Genetic Diversity

4.7.2Quantitative Investigation

5Energy Transfers (A2 only)

5.1Photosynthesis

5.1.1Overview of Photosynthesis

5.1.2Photoionisation of Chlorophyll

5.1.3Production of ATP & Reduced NADP

5.1.4Cyclic Photophosphorylation

5.1.5Light-Independent Reaction

5.1.6A-A* (AO3/4) - Photosynthesis Reactions

5.1.7Limiting Factors

5.1.8Photosynthesis Experiments

5.1.9End of Topic Test - Photosynthesis

5.1.10A-A* (AO3/4) - Photosynthesis

5.2Respiration

5.2.1Overview of Respiration

5.2.2Anaerobic Respiration

5.2.3A-A* (AO3/4) - Anaerobic Respiration

5.2.4The Link Reaction

5.2.5The Krebs Cycle

5.2.6Oxidative Phosphorylation

5.2.7Respiration Experiments

5.2.8End of Topic Test - Respiration

5.2.9A-A* (AO3/4) - Respiration

5.3Energy & Ecosystems

5.3.1Biomass

5.3.2Production & Productivity

5.3.3Agricultural Practices

5.4Nutrient Cycles

5.4.1Nitrogen Cycle

5.4.2Phosphorous Cycle

5.4.3Fertilisers & Eutrophication

5.4.4End of Topic Test - Nutrient Cycles

5.4.5A-A* (AO3/4) - Energy,Ecosystems&NutrientCycles

6Responding to Change (A2 only)

6.1Nervous Communication

6.1.1Survival

6.1.2Plant Responses

6.1.3Animal Responses

6.1.4Reflexes

6.1.5End of Topic Test - Reflexes, Responses & Survival

6.1.6Receptors

6.1.7The Human Retina

6.1.8Control of Heart Rate

6.1.9End of Topic Test - Receptors, Retina & Heart Rate

6.2Nervous Coordination

6.2.1Neurones

6.2.2Action Potentials

6.2.3Speed of Transmission

6.2.4End of Topic Test - Neurones & Action Potentials

6.2.5Synapses

6.2.6Types of Synapse

6.2.7Medical Application

6.2.8End of Topic Test - Synapses

6.2.9A-A* (AO3/4) - Nervous Comm&Coord

6.3Muscle Contraction

6.3.1Skeletal Muscle

6.3.2Sliding Filament Theory

6.3.3Contraction

6.3.4Slow & Fast Twitch Fibres

6.3.5End of Topic Test - Muscles

6.3.6A-A* (AO3/4) - Muscle Contraction

6.4Homeostasis

6.4.1Overview of Homeostasis

6.4.2Blood Glucose Concentration

6.4.3Controlling Blood Glucose Concentration

6.4.4End of Topic Test - Blood Glucose

6.4.5Primary & Secondary Messengers

6.4.6Diabetes Mellitus

6.4.7Measuring Glucose Concentration

6.4.8Osmoregulation

6.4.9Controlling Blood Water Potential

6.4.10ADH

6.4.11End of Topic Test - Diabetes & Osmoregulation

6.4.12A-A* (AO3/4) - Homeostasis

7Genetics & Ecosystems (A2 only)

7.1Genetics

7.1.1Key Terms in Genetics

7.1.2Inheritance

7.1.3Linkage

7.1.4Multiple Alleles & Epistasis

7.1.5Chi-Squared Test

7.1.6End of Topic Test - Genetics

7.1.7A-A* (AO3/4) - Genetics

7.2Populations

7.2.1Populations

7.2.2Hardy-Weinberg Principle

7.3Evolution

7.3.1Variation

7.3.2Natural Selection & Evolution

7.3.3End of Topic Test - Populations & Evolution

7.3.4Types of Selection

7.3.5Types of Selection Summary

7.3.6Overview of Speciation

7.3.7Causes of Speciation

7.3.8Diversity

7.3.9End of Topic Test - Selection & Speciation

7.3.10A-A* (AO3/4) - Populations & Evolution

7.4Populations in Ecosystems

7.4.1Overview of Ecosystems

7.4.2Niche

7.4.3Population Size

7.4.4Investigating Population Size

7.4.5End of Topic Test - Ecosystems & Population Size

7.4.6Succession

7.4.7Conservation

7.4.8End of Topic Test - Succession & Conservation

7.4.9A-A* (AO3/4) - Ecosystems

8The Control of Gene Expression (A2 only)

8.1Mutation

8.1.1Mutations

8.1.2Effects of Mutations

8.1.3Causes of Mutations

8.2Gene Expression

8.2.1Stem Cells

8.2.2Stem Cells in Disease

8.2.3End of Topic Test - Mutation & Gene Epression

8.2.4A-A* (AO3/4) - Mutation & Stem Cells

8.2.5Regulating Transcription

8.2.6Epigenetics

8.2.7Epigenetics & Disease

8.2.8Regulating Translation

8.2.9Experimental Data

8.2.10End of Topic Test - Transcription & Translation

8.2.11Tumours

8.2.12Correlations & Causes

8.2.13Prevention & Treatment

8.2.14End of Topic Test - Cancer

8.2.15A-A* (AO3/4) - Gene Expression & Cancer

8.3Genome Projects

8.3.1Using Genome Projects

8.4Gene Technology

8.4.1Recombinant DNA

8.4.2Producing Fragments

8.4.3Amplification

8.4.4End of Topic Test - Genome Project & Amplification

8.4.5Using Recombinant DNA

8.4.6Medical Diagnosis

8.4.7Genetic Fingerprinting

8.4.8End of Topic Test - Gene Technologies

8.4.9A-A* (AO3/4) - Gene Technology

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