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AH Biology > 1.1; Laboratory Techniques (new) > Flashcards

1.1; Laboratory Techniques (new) Flashcards

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1
Q

Anything that can cause harm or injury to people or that can cause ecological damage if it is released from the laboratory.

A

Hazard

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2
Q

Hazard

A

Anything that can cause harm or injury to people or that can cause ecological damage if it is released from the laboratory.

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3
Q

three types of hazards

A

substances

organisms

equipment

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4
Q

substances

organisms

equipment

A

three types of hazards

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5
Q

hazards: substances

A

toxic chemicals which build up in the body and cause damage

corrosive chemicals which can cause severe burns to exposed skin

flammable substances which can easily catch fire and potentially burn people

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6
Q

hazards: organisms

A

plants and animals may release substances that damage humans with contact

pathogenic organisms can cause deadly diseases to humans

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7
Q

hazards: equipment

A

sharp objects have the risk of cutting

mechanical equipment can present hazards if not used correctly

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8
Q

risk

A

the likelihood of harm arising from exposure to a hazard

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9
Q

the likelihood of harm arising from exposure to a hazard

A

risk

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10
Q

risk assessment

A

identifying control measures to minimise the risk posed by the hazard

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11
Q

what do control measures do to risks and hazards

A

do not remove the hazard, rather they reduce the risk down to an acceptable level.

risk assessment can only minimise risk, not remove it

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12
Q

three categories of control measures

A

appropriate handling techniques
- using tongs or oven mitts to handle hot objects

protective clothing and equipment
- safety glasses, PPE, gloves, fume cupboards

aseptic technique
- prevent exposure to, and escape of, a pathogenic organism

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13
Q

appropriate handling techniques
- using tongs or oven mitts to handle hot objects

protective clothing and equipment
- safety glasses, PPE, gloves, fume cupboards

aseptic technique
- prevent exposure to, and escape of, a pathogenic organism

A

categories of control measures

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14
Q

hazards: toxic chemicals examples

A

lead ethanoate is toxic as the lead ions can bioaccumulate in the body over long periods of time and act as a non competitive inhibitor of many enzymes

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15
Q

lead ethanoate is toxic as the lead ions can bioaccumulate in the body over long periods of time and act as a non competitive inhibitor of many enzymes

A

hazards: substances

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16
Q

hazards: corrosive chemicals examples

A

concentrated sulphuric acid can cause severe burns to exposed skin

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17
Q

hazards: corrosive chemicals examples

A

concentrated sulphuric acid can cause severe burns to exposed skin

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18
Q

concentrated sulphuric acid can cause severe burns to exposed skin

A

substances: corrosive chemicals

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19
Q

hazards: flammable substances example

A

ethanol is highly flammable at room temperature

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20
Q

ethanol is highly flammable at room temperature

A

hazards: flammable substances example

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21
Q

hazards: organisms example

A

new zealand flatworm has mucus with protease enzymes that irritates human skin

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22
Q

new zealand flatworm has mucus with protease enzymes that irritates human skin

A

hazards: organisms example

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23
Q

hazards: equipment example

A

the lid of a centrifuge must not be opened until the spinning has stopped as the rapidly moving rotor could seriously injure if touched

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24
Q

the lid of a centrifuge must not be opened until the spinning has stopped as the rapidly moving rotor could seriously injure if touched

A

hazards: equipment example

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25
Q

how to ensure accuracy when making dilutions

A

use appropriate measuring methods like scales, measuring cylinders, pipettes or autopipettes

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26
Q

linear dilution differs by…..

A

an equal interval

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27
Q

example of linear dilution

A

0.1, 0.2, 0.3, 0.4..

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28
Q

how to make linear dilution

A

add different volumes of stock solution to different volumes of solvent

each concentration is made individually so any measurement errors affect only one concentration

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29
Q

add different volumes of stock solution to different volumes of solvent

each concentration is made individually so any measurement errors affect only one concentration

A

how to make linear dilution

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30
Q

log dilution consists of..

A

a range of different dilutions that differ by a constant proportion

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31
Q

a range of different dilutions that differ by a constant proportion

A

log dilution series

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32
Q

log dilution example

A

10^-1

10^-2

10^-3

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33
Q

how to make a log dilution series

A

each dilution acts as a stock for the subsequent dilution.

each concentration depends on those made before and any earlier measurement errors are compounded in later dilutions.

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34
Q

each dilution acts as a stock for the subsequent dilution.

each concentration depends on those made before and any earlier measurement errors are compounded in later dilutions.

A

log dilution series

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35
Q

what does a colorimeter measure

A

the concentration of a pigment in a solution

or

the turbidity of liquid

or

the density of cells in a culture

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36
Q

what measures…

the concentration of a pigment in a solution

or

the turbidity of liquid

or

the density of cells in a culture

A

colorimeter

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37
Q

how does a colorimeter work

A

by illuminating a small sample of the test substance, held in a small transparent cuvette and electronically recording how much light is absorbed by the sample.

a suitable wavelength filter is used so that the concentration of a coloured solution can be determined.

for each experiment, the machine is calibrated using a blank cuvette containing solvent only which acts as a baseline for each comparison

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38
Q

what works by illuminating a small sample of the test substance, held in a small transparent cuvette and electronically recording how much light is absorbed by the sample.

a suitable wavelength filter is used so that the concentration of a coloured solution can be determined.

A

colorimeter

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39
Q

colorimeter: measuring turbidity

A

a denser sample will show a lower degree of transmission

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40
Q

buffer

A

are aqueous solutions that show very little variation in their pH despite the addition of acids or alkalis

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41
Q

are aqueous solutions that show very little variation in their pH despite the addition of acids or alkalis

A

buffer

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42
Q

Using buffers in lab experiments

A

Selected so that the pH of the reaction mixture can be kept constant

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43
Q

Centrifugation

A

A method for separating materials in suspension according to their differing density

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43
Q

A method for separating materials in suspension according to their differing density

A

Centrifugation

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44
Q

How does a centrifuge work

A

The material being studied is rotated in a centrifuge tube at between 2000 to 120 000 revolutions per minute

The resulting g force causes the constituents to separate.

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45
Q

The material being studied is rotated in a centrifuge tube at between 2000 to 120 000 revolutions per minute

The resulting g force causes the constituents to separate.

A

How a centrifuge works

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46
Q

The products of centrifugion

A

More dense = form a pellet for disposal

Less dense = liquid above called the supernatant

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47
Q

Supernatant

A

The less dense liquid fraction above

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48
Q

The less dense liquid fraction above

A

Supernatant

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48
Q

In paper and thin layer chromatography, what can amino acids and sugars be separated based on

A

Solubility

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48
Q

What can paper and thin layer chromatography separate based on solubility

A

Amino acids or sugar

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49
Q

Affinity chromatography

A

Technique for the separation of one specific soluble protein from a mixture

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49
Q

Technique for the separation of one specific soluble protein from a mixture

A

Affinity chromatography

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50
Q

Gel electrophoresis

A

Uses current flowing through a buffer to separate either proteins or nuclei acid

50
Q

Uses current flowing through a buffer to separate either proteins or nuclei acid

A

Gel electrophoresis

50
Q

What can affect the rate which any macromolecule migrated through a gel

A

Size and charge

51
Q

IEP

A

Isoelectric point

52
Q

What can size and charge affect in gel electrophoresis

A

The rate at which any macromolecule migrated through a gel

53
Q

What is IEP

A

The pH at which the protein has no net charge

54
Q

The pH at which the protein has no net charge

A

Isoelectric point

55
Q

What is there above or below the Isoelectric point of a protein

A

There will either be a majority of positive or negative charges at the surface of the protein and the protein will be soluble

56
Q

There will either be a majority of positive or negative charges at the surface of the protein and the protein will be soluble

A

Above or below the IEP

57
Q

When the protein has a charge, what happens

A

Water molecules interact with the charges

Keeping the protein in suspension

58
Q

_____ molecules interact with the charges

Keeping the protein in suspension

A

Water

59
Q

What does the neutral charge at the IEP allow for

A

The protein to form a solid and precipitate out of the solution

60
Q

what means that protein to form a solid and precipitate out of the solution

A

The neutral charge at IEP

61
Q

What can we use to separate proteins from each other

A

IEP

62
Q

Immunoassay techniques

A

Powerful tools in laboratory studies for the detection and identification of specific proteins

63
Q

Powerful tools in laboratory studies for the detection and identification of specific proteins

A

Immunoassay techniques

64
Q

What is utilised in antibody techniques

A

The ability of antibodies to bind to specific antigens

65
Q

What are antibodies

A

Y shaped globular proteins produced by B lymphocytes as part of the immune response of a vertebrate

66
Q

Y shaped globular proteins produced by B lymphocytes as part of the immune response of a vertebrate

A

Antibodies

67
Q

What does each B lymphocyte produce

A

One specific antibody

68
Q

What each produces one specific antibody

A

Each B lymphocyte

69
Q

What does each specific antigen bind to

A

Antibody

69
Q

What does each specific antibody bind to

A

One specific antigen

70
Q

What does binding of antibody to antigen cause

A

The antigen becomes harmless

71
Q

How to get a serum containing antibodies

A

Harvest from the blood of animals that have been exposed to a particular antigenic material

After time, blood is removed and antibodies are separated with centrifugion

71
Q

What are produced in the spleen and bone marrow

A

B lymphocytes

71
Q

Where are B lymphocytes produced

A

Spleen and bone marrow

72
Q

What can you from the blood of animals that have been exposed to a particular antigenic material

After time, blood is removed and antibodies are separated with centrifugion

A

Serum containing antibodies

73
Q

What is each antibody made of

A

A single B lymphocyte clone

74
Q

Serum of antibodies made with many different antibodies

A

Polyclonal antibody

75
Q

Polyclonal antibody

A

Serum of antibodies made with many different antibodies

76
Q

Monoclonal antibody

A

Stock of antibodies with the same specify

77
Q

Stock of antibodies with the same specificity

A

Monoclonal antibody

78
Q

An antibody specific to the protein antigen is linked to what

A

A chemical label

79
Q

What is a chemical label linked to

A

An antibody specific to the protein antigen

80
Q

What is the chemical label antibody’s are linked to

A

Usually a reporter enzyme producing a colour change

But chemiluminescence, fluorescence and other reporters can be used

81
Q

Usually a reporter enzyme producing a colour change

But chemiluminescence, fluorescence and other reporters can be used

A

What is a chemical label of an antibody

82
Q

What does immunoassay techniques sometimes use

A

A specific antigen to detect the presence of antibodies

83
Q

What is important when manufacturing antibodies

A

They are identical and bind to the exactly the same feature of the antigen

84
Q

How to produce pure monoclonal antibodies

A

A single line of lymphocytes must be grown, each secreting the same antibody

85
Q

What are monoclonal antibodies used in

A

Diagnosis and detection of disease

86
Q

What can be used for the diagnosis and detection of disease

A

Monoclonal antibodies

87
Q

What are immunoassay techniques used for

A

To detect and identify specific proteins

88
Q

what is western blotting used for

A

Identifying specific proteins that have been separated using gel electrophoresis

88
Q

What are used to detect and identify specific proteins

A

Immunoassay techniques

88
Q

what is used to identify specific proteins that have been separated using SDS-PAGE gel electrophoresis

A

Western blotting

89
Q

What happens to separated proteins from SDS-PAGE electrophoresis

A

Transferred onto a solid medium

90
Q

What does western blotting happen after

A

SDS PAGE gel electrophoresis

90
Q

What can you do with western blotting

A

Identify proteins using specific antibodies that have reporter enzymes attached

91
Q

how can you identify proteins using specific antibodies that have reporter enzymes attached

A

Western blotting

91
Q

Bright field microscopy is commonly used…

A

To observe whole organisms, parts of organisms, thin sections of dissected tissue, or individual cells

92
Q

What is used to observe whole organisms, parts of organisms, thin sections of dissected tissue, or individual cells

A

Bright field microscopy

93
Q

What does fluorescence microscopy use and do

A

Specific fluorescent labels to bind to and visualise certain molecules or structures within cells or tissues

94
Q

What uses specific fluorescent labels to bind to and visualise certain molecules or structures within cells or tissues

A

Fluorescence microscopy

94
Q

What does aseptic technique do

A

Eliminates unwanted microbial contaminants when culturing micro organisms or cells

95
Q

What eliminates unwanted microbial contaminants when culturing micro organisms or cells

A

Aseptic technique

96
Q

What does aseptic technique involve

A

The sterilisation of equipment and culture media by heat or chemical means and subsequent exclusion of microbial contaminants

97
Q

What can a microbial culture be started using

A

An inoculum of microbial cells on an agar medium or in a broth with suitable nutrients

98
Q

What is s

A
99
Q

What is started with an inoculum of microbial cells on an agar medium or in a north with suitable nutrients

A

A microbial culture

100
Q

Specific culture medias

A

Promote the growth of specific types of cells and microbes

101
Q

what promotes the growth of specific types of cells and microbes

A

Culture media

102
Q

What are animal cells growth in

A

Medium containing growth factors from serum

103
Q

How many times can primary cell lines divide in culture

A

A limited number of times

104
Q

How many times can tumour cell lines divide in culture

A

Unlimited divisions

105
Q

What cells can have unlimited divisions in culture

A

Tumour cell lines

106
Q

What cells can have limited divisions in culture

A

Primary cell lines

107
Q

What allows for the number of colony forming units to be counted and the density of cells in the culture estimated

A

plating out a liquid microbial culture on solid media allow for

107
Q

What does plating out a liquid microbial culture on solid media allow for

A

The number of colony forming units to be counted and the density of cells in the culture estimated

108
Q

What is serial dilution often needed to achieve

A

A suitable colony count

109
Q

What can be needed to get a suitable colony count

A

Serial dilution

110
Q

What is needed to identify and count viable cells

A

Viral staining

111
Q

What does an assay do

A

determined the presence or concentration of a substance in a mixture

112
Q

What can viral staining be used for

A

Identifying and counting viable cells

113
Q

Immunoassay

A

Uses a monoclonal antibody to determine the presence or concentration of a substance in a mixture

114
Q

determined the presence or concentration of a substance in a mixture

A

Assay

115
Q

Uses a monoclonal antibody to determine the presence or concentration of a substance in a mixture

A

Immunoassay

116
Q

Damage of bacteria or fungal contamination

A

Outcompete and spoil a culture of slower growing cells

117
Q

what will spoil a culture of slower growing cells

A

Bacterial or fungal contamination

118
Q

what are growth factors

A

Proteins that promote cell growth and proliferation

They are essential for the culture of most animal cells

AH Biology (3 decks)

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