2.1.6
Methods of Studying Cells
Magnification and Resolution
Magnification and Resolution
Two parameters that are important in microscopy are magnification and resolution.
Magnification
Magnification
- Magnification is the process of enlarging an object in appearance.
- The image size is how big the object appears to be in a picture or drawing, which will be in milimeters (mm).
- The actual size is often given in micrometers (µm) - units must be converted so that they are the same.
Calculating magnification
Calculating magnification
- The equation for magnification is:
- Magnification = size of image ÷ size of real object
- E.g. A mitochondrion is 20 µm long. An image of the mitochondrion is measured as 20 mm long. What is the magnification?
- Magnification = 20,000 µm ÷ 20 µm = 1,000x
Resolution
Resolution
- Resolution is the ability of a microscope to distinguish two adjacent structures as separate.
- The higher the resolution, the better the clarity and detail of the image.
Cell Fractionation
Cell Fractionation
Cell fractionation separates organelles according to size to allow them to be studied in an electron microscope. The steps involved are:
1) Homogenisation
1) Homogenisation
- The tissue sample is homogenised using a blender to break the cells.
- The tissue sample must be kept in specific conditions:
- Ice cold (reduces enzyme activity that might damage organelles).
- Isotonic solution (prevents osmosis that could shrink or burst organelles).
- No osmosis takes place in isotonic solution.
- Buffered solution (keeps pH constant and avoids damaging the protein structures).
2) Filtration
2) Filtration
- The tissue sample is filtered into tubes through a gauze.
- The gauze separates larger components from the small, organelles.
- The organelles are filtered into tubes to be fractionated using ultracentrifugation.
3) Ultracentrifugation
3) Ultracentrifugation
- The samples are spun at a low speed in a centrifuge.
- Each tube must be balanced with another tube directly opposite for the centrifuge to work properly.
- Centrifugation separates the sample into fractions.
- Heavier organelles are forced to the bottom of the tube.
- Lighter organelles move towards the top.
- Cell debris (e.g. cell walls) forms a pellet at the bottom of the tube, leaving the supernatant (a liquid) above it that contains the organelles.
4) Ultracentrifugation
4) Ultracentrifugation
- The supernatant is poured off and centrifuged at a higher speed to separate the next heaviest organelles (the nuclei).
- This is repeated at increasingly higher speeds to separate each fraction.
5) Order of fractionation
5) Order of fractionation
- The process of repeated ultracentrifugation produces fractions of cell organelles from heaviest to lightest. This order is:
- Nucleus.
- Chloroplasts.
- Mitochondria.
- Lysosomes.
- Endoplasmic reticulum.
- Ribosomes.
1Biological Molecules
1.1Monomers & Polymers
1.2Carbohydrates
1.3Lipids
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
1.6ATP
1.7Water
1.8Inorganic Ions
2Cells
2.1Cell Structure
2.2Mitosis & Cancer
2.3Transport Across Cell Membrane
2.4Cell Recognition & the Immune System
3Substance Exchange
3.1Surface Area to Volume Ratio
3.2Gas Exchange
3.3Digestion & Absorption
4Genetic Information & Variation
4.1DNA, Genes & Chromosomes
4.2DNA & Protein Synthesis
4.3Mutations & Meiosis
4.4Genetic Diversity & Adaptation
4.5Species & Taxonomy
4.6Biodiversity Within a Community
4.7Investigating Diversity
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
5.3Energy & Ecosystems
6Responding to Change (A2 only)
6.1Nervous Communication
6.2Nervous Coordination
6.3Muscle 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
7.2Populations
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
8The Control of Gene Expression (A2 only)
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
Jump to other topics
1Biological Molecules
1.1Monomers & Polymers
1.2Carbohydrates
1.3Lipids
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
1.6ATP
1.7Water
1.8Inorganic Ions
2Cells
2.1Cell Structure
2.2Mitosis & Cancer
2.3Transport Across Cell Membrane
2.4Cell Recognition & the Immune System
3Substance Exchange
3.1Surface Area to Volume Ratio
3.2Gas Exchange
3.3Digestion & Absorption
4Genetic Information & Variation
4.1DNA, Genes & Chromosomes
4.2DNA & Protein Synthesis
4.3Mutations & Meiosis
4.4Genetic Diversity & Adaptation
4.5Species & Taxonomy
4.6Biodiversity Within a Community
4.7Investigating Diversity
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
5.3Energy & Ecosystems
6Responding to Change (A2 only)
6.1Nervous Communication
6.2Nervous Coordination
6.3Muscle 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
7.2Populations
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
8The Control of Gene Expression (A2 only)
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
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