Unit 1 Lab Techniques Flashcards
Define affinity
The degree to which a substrate is attracted and tends to bind to another
Define affinity chromatography
A technique used to separate and purify proteins based on a specific binding interaction between the ligand and its binding partner
Define an antigen
A specific protein with which antibodies can bind in an immune response
Define aseptic technique
Procedures in place to prevent contamination including sterilising equipment and work surfaces
Define bright field microscopy
Microscopy commonly used to observe whole organisms, parts of organisms, thin sections of dissected tissue or individual cells
Define a buffer
A solution used to set and maintain a particular pH
Define centrifugation
A process that uses centrifugal forces to separate components of different densities in a mixture
Define a chromatography
A technique used to separate different substances; it has a stationary phase that the mobile phase moves through, carrying the substance being examined; different distances are moved by substances of differing solubilities
Define a colorimeter
A device used to measure absorbance of specific wavelength of light by a solution
Define a culture media
A nutrient rich gross medium providing their basic requirements for cell growth (amino acids glucose salt water as well as specific growth factors for animal cell lines)
Define fluorescence
The emission of light of different wavelengths to that which was absorbed
Define fluorescence microscopy
A microscopy technique that uses specific fluorescent labels to bind and visualise certain molecules or structures within cells or tissues
Define gel electrophoresis
A technique used to separate samples of nucleic acid and proteins by size; introduced to a gel they move through it due to an electric current smaller fragments travel further than larger fragments
Define growth factors
Proteins that promotes cell growth and proliferation
Define a haemocytometer
Microscopic grid used to estimate the total number of cells within a sample (originally used to count red blood cells)
Define a hazard
Anything that poses a potential risk or threat to an individual or the environment
Define immunoassay
Technique used to detect and identify specific proteins; antibodies linked with reporter enzymes e.g. calls a colour change in the presence of specific antigen
Define inoculum
Starting material used to grow a culture from for example a bacteria cell culture
Define Isoelectric point (IEP)
The pH at which a soluble protein has no net charge and will precipitate out a solution
Fine linear dilution series
A series of delusions that differ by a constant proportion
Define log dilution series
A series of solutions that differ by equal intervals
Define monoclonal antibodies
Stocks of identical antibodies that are specific to a particular antigen
Define native gel electrophoresis
Does not contain SDS And is not denature proteins so proteins are separated by sheet size and charge
Define pathogenic
Causing disease
Define primary cell lines
A culture of sells ice deleted directly from an animal or plant tissue they have an infinite lifespan and expansion capacity
Define reporter enzymes
An enzyme linked to an Ab specific to a protein antigen they are used and immunoassay techniques
Define SDS page
Electrophoresis in which gel contains SDS which gives all molecules and equally negative charge and denatures them separating proteins by size alone
Define serum
Vitally important as source of growth factors hormones leopards and minerals for culture cells
Define standard curve
A graph that can be used to determine the concentration of an unknown solution
Define supernatant
The liquid that lives above a solid residue are Pella in the centrifugation
Define turbidity
A measure of the degree to which a fluid loses its transparency due to the presence of suspended particles or cells in suspension
Define vital staining
A technique In which a harmless die is used to staying in their living tissues are dead cells for Microscopical observation to allow viable cell count to be made
Define western blotting
An analytical technique used to identify and locate specific proteins in a sample of tissue homogenete or extract based on their ability to bind to specific antibodies
What is the difference between a typical culture media and a complex culture media
A typical culture media contains what are salts amino acids vitamins and glucose
Complex culture media has growth factors that promote so growth and proliferation
Describe the process of western blotting
The proteins are transferred from a sale to a solid medium
The membrane is blocked to prevent antibodies from binding to the wrong part of the membrane
Antibody is added and then washed off
A secondary label antibody is added and then washed off
Results are obtained by chemiluminescence
Explain the difference between absorbance transmission and turbidity
Turbidity is a degree to which a fluid loses its transparency due to the presence of suspended particles in suspension
Absorbance the quality of light absorbed by solution
Transmission is the quantity of light that passes through a solution
Why is fluorescent labelling of antibodies useful
Fluorescent labelling for antibodies allows for visualising proteins
Sensitive At low concentrations are stable over long periods of time and do not interfere with the function of the target molecules
How and why is brightfield microscopy carried out
A sample is mounted on a slide and illuminated from bellow
Samples are staying before viewing using a Brakefield my scope to increase contrast contrast
Used to examine whole organisms/parts of them
How and why is present field microscopy carried out
Is used image specific features of small specimens such as microbials
Use a specific for us and labels to bind and visualise certain molecules
What is the haemocytometer used for
It is accounting chamber device originally designed and used for counting blood cells
Why would you use tumour cell lines instead of primary cell lines for culturing cells
Tumour cells can perform unlimited divisions (immortal sales) whereas primary cells die after certain amount of divisions (mortal cells)
Name the components of a complex culture media required by animal cells were they used for
Proteins that promote self growth and proliferation
Use for the cultivation of bacterial pathogens and other fastidious bacteria
Why is vital staining used to estimate viable cell count
Viable stealing allowed viable sells beer then divide and Celko to be before him where as only answered in sales are counted as die is only taking up by dead cells
Give three health and safety considerations for procedures
Where are your hair up
Wear goggles
Where lab coats
Give a brief message for linear and log dilutions
Lock delusions measure a range of solutions that differ five constant for portion each solution access the stock for the next
Linear delusions differ by equal intervals and add measures of volume to the stock solution
Why are standard curves used
As a standard glass can be used to determine the concentration of an unknown sample
Why are buffers used
To keep the pH of a reaction constant
How do you calorimeters work and why are they used
A colorimeter is used to measure the quantity of like the sample absorbs it transmits operate by passing a beam of light through a sample and measuring the light intensity reaching the detector
How does the centrifugal separate materials
I love separation of materials to the use of a rotor that spins in a centrifuge a centrifugal force is applied to each particle the sample the particle will then sediment at the rate that the rate proportional to the force that was applied
Describe both methods and purpose of protein electrophoresis
Native jealous every proteins by their size shape and charge they do this by ensuring they do not denature the molecule
SDS drill separate proteins by size alone proteins are heated in the presence of a detergent
Describe the method and purpose of isoelectric focusing
A pH gradient is that up alone a tube of gel using a special offer a mixture and then electrical field is applied
Describe how are use immunoassay to diagnose conditions such as HIV
An antibody test checked for antibodies to the HIV virus the enzyme like with a Venus absorbent is ELIZA which is used for the detection of HIV antibody and antigen
If the antigen is present the antibody binds if the substrate is added and a colour change this suggests HIV virus
Name three types of chromatography and briefly describe the methods and purposes
Paper separation of mixtures based on their solubility is the speed this all your travels along the chromatograms depends on at solubility
Thin-layer chromatography is the same except it’s performed on a sheet of glass or plastic which is coated with a layer of absorbing material usually gel or Celulose
Infinity chromatography separate target proteins are a mixture soluble target proteins in a mixture will become attached to them as the mixer passed down the column molecules with no affinity are washed out target protein move from the column
