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AH Biology > Lab Techniques for Biologists > Flashcards

Lab Techniques for Biologists Flashcards

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1
Q

Give examples of substances covered by COSHH?

A 
Chemicals
Products containing chemicals
Fumes
Dusts
Vapours
Biological agents
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2
Q

What does COSHH stand for?

A

Control of Substances hazardous to health

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3
Q

What does PPE stand for?

A

Personal Protective Equipment

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4
Q

Give some examples of PPE

A

Safety goggles
Lab coat
Gloves
Closed toe shoes

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5
Q

What does an autopipette allowed to be measured?

A

Small volumes of liquid accurately

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6
Q

What are burettes used for?

A

Dispense precise volumes of liquid reagants

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7
Q

What happens to the burette before it is used?

A

Rinsed with the liquid to be used

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8
Q

Where are readings taken from?

A

Bottom of the Meniscus

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9
Q

What is repeated dilution from a stock solution known as?

A

Serial Dilution

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10
Q

What type of dilution is it if each step is a tenfold dilution?

A

Logarithmic Dilution

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11
Q

The concentration of a pigmented compound can be quantified using a?

A

Colorimeter

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12
Q

How does a colorimeter work?

A

Passing a light beam at a specific wavelength through a cuvette containing a sample solution

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13
Q

How do you calibrate a colorimeter?

A

Place a calibration solution(usually distilled water) into a cuvette, place in colorimeter and press calibrate, providing a user with a 0 absorbance reading

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14
Q

What is a pH buffer?

A

Solution whose pH changes very little when a small amount of acid/base is added to it

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15
Q

Give one method to determine the unknown concentration of a solution

A

Standard Curve

Titration

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16
Q

What seperates substances out in centrifugation?

A

Size and Density

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17
Q

What is the liquid that remains above the pellet called in centrifugation?

A

Supernatant

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18
Q

What does protein electrophoresis seperate proteins based on?

A

Charge and/or size/shape

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19
Q

Give one form of protein electrophoresis

A

SDS page

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20
Q

Do smaller proteins travel furthur or less than bigger proteins in protein electrophoresis?

A

Further

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21
Q

What does seperation of proteins depend on if native gels are used?

A

protein charge and shape.

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22
Q

What do proteins do at their isoelectric point?

A

Overall neutral charge, precipitate out of solution

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23
Q

What charge do proteins carry below their isoelectric point?

A

Positive

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24
Q

How does isoelectric focussing work?

A

pH gradient is set up on a gel

Protein loaded in and moves until it reaches its isoelectric point where it forms a band

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25
Q

What is Chromatography?

A

Set of Techniques which separate the components of a mixture

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26
Q

What are the three types of chromatography?

A

Paper, thin-layer and Affinity

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27
Q

What is the stationary phase in paper chromatography?

A

Strip of chromatography paper

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28
Q

What is the stationary phase in thin layer chromatography?

A

Strip of absorbent material such as silica gel

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29
Q

What does affinity chromatography rely on?

A

Binding interactions of a protein and a ligand

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30
Q

Where is the ligand found in affinity chromatography?

A

Immobilised in an inert support

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31
Q

What is immunoassay?

A

Test which uses antibodies to detect the presence and concentration of a protein within a solution

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32
Q

What does immunoassay rely on?

A

Specificity of Antibodies

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33
Q

What allows scientists to detect when binding has occurred in an immunoassay test?

A

Each antibody must be linked to a detectable label

34
Q

What does a reporter enzyme do?

A

Causes a colour change in the presence of a specific antigen

35
Q

What does ELISA stand for?

A

Enzyme linked immunosorbent assay

36
Q

What is an ELISA?

A

Analytical technique which uses antibodies to detect the presence of an antigen in a solution

37
Q

What are three types of ELISA?

A

Direct, Indirect, Sandwich

38
Q

What is the difference between direct and indirect ELISA?

A

Direct - Primary antibody binds to antigen and has a reporter protein on it
Indirect - Primary antibody binds to antigen then secondary antibody binds with a reporter enzyme

39
Q

What happens during Sandwich ELISA?

A

Capture antibody bound to surface of multiwell plate
Antigen is added and allowed to bind
Primary antibody added and binds
Secondary antibody, linked to a reporter enzyme added and binds to primary antibody

40
Q

Give examples of uses of ELISA

A

Detection of HIV
Detection of food allergies
Screening for certain types of drugs

41
Q

What does Protein Blotting allow?

A

Identify specific proteins seperated by gel electrophoresis

42
Q

What is the first step in protein blotting?

A

Proteins are separated using gel electrophoresis and transferred to a membrane

43
Q

How is the membrane probed for the protein of interest?

A

Using a specific antibody linked to a detectable label

44
Q

Give examples of applications of protein blotting

A

Identify specific proteins which are present in a cell sample
Detection of HIV and hepatitis B

45
Q

What is the difference between ELISA and Immunohistochemistry?

A

ELISA - using antibodies to detect antigens in a specific solution
Immunohistochemistry - using antibodies to detect antigens in a tissue sample

46
Q

What is Immunhistochemistry typically used for?

A

Diagnosis of Diseases such as Cancer

47
Q

How does immunohistochemistry work?

A

Samples of tissue are very thinly sliced and mounted onto a slide
Probed for particular protein using an enzyme linked antibody

48
Q

What is an advantage of an immunohistochemistry test?

A

Capable of showing exactly where a certain protein is being expressed in a sample

49
Q

What is combined to form hybridomas?

A

B lympochytes and Myeloma Cells

50
Q

What is used to fuse B lympochytes and Myeloma cells?

A

Polyethlyene glycol (PEG)

51
Q

Where are B lympochytes removed from?

A

Spleen of an animal

52
Q

What is the name of the medium that hybridoma cells are cultured in?

A

Hypoxanthine Aminopterin Thymidine (HAT)

53
Q

Why do unfused myeloma cells die?

A

Cannot survive in the HAT medium

54
Q

Why do unfused B lympochytes die?

A

Cannot survive in the culture

55
Q

What does Fluorescence microscopy allow?

A

Particular protein structures to be visualised

56
Q

What is a fluorescent molecule?

A

One which absorbs a specific wavelength of light then emits a different (longer) wavelength

57
Q

How does Fluorescence microscopy work?

A

Specific proteins have fluorescent markers added to them
Cells then placed on a slide
Protein structures visualised using a fluorescence microscope

58
Q

What is immunofluoresecence?

A

Antibodies used to fluorescently tag protein structures

59
Q

Describe how immunofluoresecence works?

A

Primary antibody, specific to the protein being visualised is introduced
A second antibody, attached to a fluorescent tag which binds to the primary antibody

60
Q

What does a haemocytometer allow?

A

Estimation of the concentration of cells in a sample to be made

61
Q

What border can cells be touching to be counted?

A

Top and right border

62
Q

What can the reliability of a glass haemocytometer be effected by?

A

Incorrect placement of the cover slip

63
Q

Give a benefit of disposable haemocytometers?

A

Limit exposure to infectious materials

64
Q

Give examples of things a typical culture media contains

A 
Water
Salts
Amino Acids
Vitamins
Glucose
65
Q

What do all animal cell cultures require?

A

Media containing growth factors

66
Q

Where do the growth factors come from?

A

Serum

67
Q

What is the inoculum?

A

Cells that have been used to inoculate culture media

68
Q

What are the two different cells that can make up inoculum?

A

Cells released from source tissue using proteolytic enzymes

Explants (small pieces of tissue)

69
Q

What is a polyclonal serum?

A

A serum containing antibodies that have been harvested from the blood of animals exposed to a particular antigenic material

70
Q

What are antibodies?

A

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

71
Q

What are antibodies used for?

A

Detect and Identify specific proteins

72
Q

What does fluorescent labelled protein blotting allow a scientist to identify?

A

Specific proteins that were seperated using gel electrophoresis

73
Q

What are the steps in fluorescent labelled protein blotting?

A
  1. Protein blotted to membrane
  2. Membrane flooded with fluorescent labelled monoclonal antibodies
  3. When exposed to certain wavelengths allowing precise location of target proteins
74
Q

What allows precise location of proteins in fluorescent labelled protein blotting?

A

Fluorescent Labelled Protein Blot

75
Q

What is histochemistry?

A

Study of tissues using stains and microscopy

76
Q

What is fluorescent Immunohistochemical Staining?

A

Used to visualize distribution of specific cellular components in live cell

77
Q

What does separation rely on if non native gels are used in protein electrophoresis?

A

Size

78
Q

State two types of biological material that can be viewed by bright field microscopy

A
  • Thin sections of tissue

- Parts of an organism

79
Q

How are cells detached from tissues for cell lines?

A

Using proteolytic enzymes

80
Q

What are Explants?

A

Small pieces of plant tissue placed on solid medium

81
Q

What can be used to trigger embryogenesis?

A

Growth Regulators

AH Biology (17 decks)

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