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A/ AH Biology: Unit 1 > Key Area 1: Laboratory Techniques for Biologists > Flashcards

Key Area 1: Laboratory Techniques for Biologists Flashcards

1
Q

What are two things that can be intrinsically harmful in a lab?

A

Chemicals and organisms

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2
Q

What three things are at risk in a lab?

A
  • people
  • environment
  • other organisms
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3
Q

What are five control measures in order of most to least preferred?

A
  • elimination: replace the hazardous substance
  • substitution: use alternatives
  • isolation: contained environment
  • education: train people
  • personal protective equipment: last line of defence
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4
Q

What are risk assessments?

A

Carried out to ensure that working conditions are safe by identifying hazards and minimising risks through implementation of control procedures

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5
Q

What are devices for measuring liquids?

A
  • burette: titrations accurate for 1-100cm3
  • pipette: 30microlitres to 2cm3
  • syringe: 0.5microlitres to more than 50cm3
  • autopipette: 1microlitre to 20cm3
  • measuring cylinder: 5-2000cm3
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6
Q

What is a serial dilution?

A

Stepwise dilution where the dilution factor at each step is constant

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7
Q

What formula is used to calculate dilutions?

A

V1C1=V2C2

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8
Q

What is a linear dilution?

A

Even quantity separations

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9
Q

What is a log dilution?

A

Increase of factor ten each time

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10
Q

What are linear solutions used for?

A

Standard curves

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11
Q

What are long solutions used for?

A

Culturing microbes

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12
Q

What is a standard curve?

A

A graph that shows known concentrations and are used to determine unknown concentrations

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13
Q

HOw can standard curve concentrations be measured?

A

Colorimeter

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14
Q

How can pH be tested in an experiment?

A
  • universal indicator
  • pH paper
  • pH probe
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15
Q

How can you control pH?

A

With a buffer solution which resists a change in its pH when small amounts of acid or base is added into it

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16
Q

How does centrifugation seperate?

A

Density

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17
Q

What is the pellet?

A

The solid that forms at the bottom of the vessel in centrifugation

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18
Q

What is the supernatant?

A

The remaining liquid in centrifugation

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19
Q

What are three types of chromatography?

A

Thin layer
Affinity
Paper

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20
Q

What is thin layer chromatography?

A

Seperate by solubility on glass or plastic as amino acids with differing solubility travel different distances up the solid support

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21
Q

What is affinity chromatography?

A

Separates by specific bindingbetween molecules, separating biochemical mixtures based on a highly specific interaction such as that between antigen and antibody

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22
Q

What is paper chromatography?

A

Separates by solubility on paper, amino acids with different solubilitys travel different distances up the paper support being used

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23
Q

What is responsible for the separation of molecules in chromatography?

A

Solubility

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24
Q

How does electrophoresis separate?

A

By charge of size

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25
How does electrophoresis work?
Proteins travel different distances through a gel with different charges at either end
26
What are two factors that effect the migration of proteins in a gel?
Charge and size
27
What is the separating factor in iso-electric focussing?
PH
28
How does iso-electric focussing work?
Protein no longer migrates at the pH where it has no overall charge and the protein precipitates out of solution
29
Where does the protein precipitate out of solution and can be identified?
ISO-electric point
30
What are antibody techniques used for?
Detecting and identifying proteins
31
Why are antibodies suitable for detecting and identifying proteins?
They fight infection by specific binding to only one protein
32
What are monoclonal antibodies?
Antibodies created from identical cells that attach to specific molecules
33
What two cells fuse together to produce monoclonal antibodies?
Myeloma and B-lymphocytes
34
What is the purpose of B-lymphocyte?
Produces the antibody
35
What is the purpose of the myeloma?
Reproduce continuously without need of control
36
What is the name given to the cell made by fusion of B-lymphocyte and myeloma?
Hybridoma
37
What is used in the fusion process to produce the hybridoma?
Polyethylene glycol (PEG)
38
What is immunoassay?
Based on antigen/antibody bonding principles and can be used to detect specific proteins
39
What are the three types of Elisa?
- direct Elisa - indirect Elisa - capture Elisa
40
What is direct Elisa?
Antibody attaches directly to protein of interest
41
What is indirect Elisa?
Secondary antibody with reporter enzyme attaches to secondary antibody that is attached to protein of interest
42
What is capture Elisa?
Primary antibody added first, then protein that attaches to primary antibody, then secondary antibody that attaches to protein, then another antibody with a reporter enzyme attaches to the secondary antibody
43
How do reporter enzymes detect specific proteins?
Observable colour changes
44
How can antibodies be labelled for detection?
Reporter enzymes or fluorophore
45
What is protein blotting?
After electrophoresis, the separates proteins are blotted onto a membrane which is treated with fluorescent malt labelled specific antibodies
46
What is immunohistochemical staining?
Tissue is stained with specific labelled antibodies
47
What is examined with brightfield microscopy?
Whole organisms, parts of organisms or thin sections of dissected tissue
48
What is fluorescence microscopy?
Detecting specific proteins that have been bound to fluorescentkt labelled antibodies
49
What is aeseptic technique?
Procedure used to ensure the environment is sterile
50
What are five ways of maintaining a sterile environment?
- sterilisation of equipment - disinfection if working area - wire loop flames before use - culture dish sealed - petri dish lid only partly opened at an angle
51
What is the innoculum?
The original stock of cells used to innoculate the culture medium
52
What is the explant?
The innoculum taken from plant or animal cell (check types notes)
53
What is a haemocytometer used for?
Calculating cell counts
54
What is trypan blue dye used for?
Staining cells
55
What type of cells does trypan blue dye stain?
Only dead cells as the living cells can push out the dye
56
What is vital stain?
Dye is only taken up by non-viable cells so viable cells are easily counted
57
What is viable cell count?
Number of living cells in a sample
58
What is non-viable cell count?
The number of dead cells in a sample
59
What is total cells count?
Number of both living and dead cells in a sample
60
What is a simple culture medium?
Allows conditions for gas exchange, a suitable pH and temperature and a suitable growing surface
61
What is a complex growth medium?
Different nutrients are added to growth media depending on what you are growing
62
What are the contents of a complex culture medium?
- growth factor - serum - glucose/food sources - pH buffers - auxins
63
What is the growth factor used for culturing plant cells?
Oxins
64
Why is serum used in a complex culture medium?
It’s a source of growth factors
65
What is proliferation?
The cell can divide
66
What is a mono layer when culturing mammal cells?
A single layer of cells flat on the top of the surface
67
What are primary cell lines?
Normal cells that only divide a certain number of times then die
68
What are cancerous cell lines?
Can grow and divide infinitely without normal control of division or need for modification

A/ AH Biology: Unit 1 (6 decks)

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