Cell structures Flashcards
What does TEM stand for?
Transmission electron microscope
What is the source of image in a TEM?
the electrons transmitting through the specimen
how is the image focussed in a TEM?
electromagnets straightening the electron beam
What is the magnification of a TEM?
x500,000-x2,000,000
What is the resolution of a TEM?
0.2-0.5nm
Can you view live specimen in a TEM and why?
no- specimen are placed in a vacuum
Is staining required in a TEM?
yes- heavy metal staining is used
What are some advantages of TEM?
- high resolution
- high magnification
What are some disadvantages of TEM?
- very expensive
- black and white images
- specimen can be damaged by electron beam
What does SEM stand for?
scanning electron microscope
What is the source of image in an SEM?
the electrons reflecting off the sides of the specimen
How are SEM microscopes focussed?
electromagnets straightening the electron beam
What is the magnification of an SEM?
x100,000-x500,000
What is the resolution of an SEM?
3-10nm
Can live specimen be used in an SEM and why?
no- they are placed in a vacuum
Is staining required in an SEM?
heavy metal staining is used
What are some advantages of SEM?
- high resolution
-high magnification - 3D imaging
What are some disadvantages of SEM?
- expensive
- black and white images
- specimen can be damaged by electron beam
What is the source of image in a light microscope?
the lamp
How is a light microscope focussed?
objective lens and eye piece lens
What is the magnification of a light microscope?
x1500-x2000
What is the resolution of a light microscope?
200nm/ 0.2um
Can you view live specimen in a light microscope and why?
yes- they are only placed on a slide and they can be suspended in water
Is staining required for a light microscope?
no- only if you are viewing cells as their organelles are colourless
What are some advantages of light microscopes?
- portable
- cheap
- easy to use
-can view live specimen
What are some disadvantages of light microscopes?
- low magnification
- low resolution
What is the source of image in a laser scanning confocal?
a laser with high light intensity scans image
How is the image focussed in a laser scanning confocal?
eye piece lens and objective lens
what is the magnification of a laser scanning confocal?
high
what is the resolution of a laser scanning confocal?
very high
can you view live specimen in a laser scanning confocal?
yes only a laser of light is used
is staining required for laser scanning confocal?
no
what are some advantages of a laser scanning confocal?
- 3D images
- non-invasive
- high resolution
- high magnification
What are some disadvantages of a laser scanning confocal?
- very expensive
- requires training
why should cover slips be added at an angle?
to prevent air bubbles
why must samples on a slide be thin?
to let light through so you can see the organelles
what are the two main groups of bacteria in gram staining called?
gram positive and gram negative
what is the process of gram staining?
1) bacteria is stained with crystal violet
2) all bacteria is stained purple
3) add iodine and stain is fixed due to reaction
4) washed in alcohol
5) gram positive will stain purple and negative will lose stain
6) stained with safranin
7) gram neg will stay red and pos will stay purple
why does gram positive stain?
- they have a thick layer of peptidoglycan in their cell walls
why does gram negative stain red?
- thinner layer of peptidoglycan
- will not retain violet stain well during the decolouring process
- cell walls are not susceptible to penicillin
who made the first microscope and how did they do it?
- Hans and Jensen
- put lenses in a tube
who also claimed they invented the first microscope?
Galilei
who first observed cells?
Robert Hooke
What are proposed theories based on?
current and accepted evidence
what is the importance of a microscope?
- allows you to view cells
- many organisms are unicellular so you cant see them without a microscope
- allowed people to discover new organisms and processes
why do images tend to have a low contrast in a light microscope?
- cells don’t absorb a lot of light
- light is below and you view from above
what is the importance of staining?
- increases contrast between components as they take up stains to different degrees
- allows organelles to be visible for identification
what is gram staining?
a technique used to differentiate between two large groups of bacteria based on their different cell wall constituents
in a wet mount, what should be the same a glass and why?
the refractive index should be the same of that of glass to prevent diffraction
what is the definition of magnification?
how many times larger an image is compared to the actual image
resolution definition
the smallest distance between two points which can still be distinguished as separate entities
what is the equation for the actual size of an image?
actual size = image size/ magnification
how is a dry mount specimen prepared?
cut into very thin slices with a sharp blade
how is a wet mount specimen prepared?
specimen suspended in water or immersion oil
how is a squash slide specimen prepared?
specimen suspended in water or immersion oil and lens tissue is used to gently press down the cover slip
how is a smear slide specimen prepared?
edge of slide is used to smear the sample
what are some examples of specimens used in dry mounts?
- hair
- insects
- pollen
what are some examples of specimens used in squash slides?
soft samples
what are some examples of specimens used in wet mounts?
- aquatic samples
- other living organisms
what are some examples of specimens used in smear slides?
- blood
- any liquid
how does diffraction and wavelength of light affect resolution?
shorter wavelength yield means higher resolution
explain why staining is needed in terms of contrast?
staining increases contrast between components as they take up stains at different degrees
give some examples of positively charged dye and what is it attracted to?
- crystal violet and methylene blue
- attracted to negatively charged material in cytoplasm
give examples of negatively charged dyes and what is it repelled by?
- nigrosin and congo red
- repelled by negatively charged cytosol
what is the purpose of fixing (in slide production)?
to preserve specimens (chemicals such as formaldehyde)
what is sectioning (in slide preparation)?
specimens are dehydrated with alcohol and placed in a wax block before thinly sliced with a microtome
what is the purpose of staining (in slide preparation)
to view different organelles as different stains show different structures
what is mounting (in slide preparation)?
specimens are secured to a microscope slide with a cover slip
what is differential staining?
staining that helps differentiate between different types of organelles
what does CLEAPSS stand for and what do they do?
- Consortium of Local Education Authorities for the Provision of Science Services
- provide support for practical work in schools
what is on a student safety sheet?
- identify specific risks
- advice on the measures to be taken to reduce risks
- action to take in emergency
why are most slides pre-prepared in schools?
- harmful nature of stains
- long complex process needed to produce high quality sections
how does penicillin affect gram positive staining?
it inhibits the peptidoglycan production, making the cells fragile
how does penicillin affect gram negative bacteria?
it has peptidoglycan between membranes so the penicillin doesn’t work aswell
what is the acid fast staining technique used for?
to differentiate species of mycobacteria from other bacteria
what dye is used in acid fast staining?
carbolfuchsin dye
what is used to carry dye into the cells in acid fast staining?
a lipid solvent
how do you calculate the number of eye piece divisions?
number of micrometres/ number of eye piece divisions
what is a scale micrometer?
a microscope slide with an accurate scale in um
- 100 divisions= 1mm
- 1 division= 0.01mm= 10um
what is an eye piece graticule?
- a glass disc with a scale on it
- inserted into a microscope eyepiece
- the scale remains the same at each magnification
how do you convert from um to nm?
x1000
how do you convert from mm to um?
x1000
how do you convert from cm to mm?
x10
what is the function of a vacuole?
- maintain turgor (correct pressure)
- selectively permeable
- plants have a permanent vacuole
what is the function of cillia?
- sensory- detect things
- movement
what is the function of the flagella?
movement
what is the function of the cell wall?
- shape
- rigidity
- strength
what is the function of centrioles?
produce spindle fibres and cytoskeleton
how thick is the plasma membrane?
7nm
what is the function of the chloroplasts?
photosynthesis
what is the function of the lysosomes?
- contains digestive enzymes
- breaks down old organelles, bacteria and foreign bodies
what is the function of ribosomes?
- proteinsynthesis
what is the function of the mitochondria?
aerobic respiration (in the cristae) which produces ATP
what is the function of the smooth endoplasmic reticulum (SER)?
synthesis and transports lipids and steroids (lipid synthesis)
what is the function of the golgi apparatus?
processes and packages vesicles
what is the structure of the nuclear envelope?
- double membrane
- has pores
what is the function of the rough endoplasmic reticulum (RER)?
- protein synthesis
- transport of proteins
what is the size of a nuclear pore?
120nm
what is the function of the nuclear envelope?
contains genetic material in a certain place
what is the function of the nucleolus?
makes RNA and ribosomes
what is the function of the nucleus?
- contains genetic material (DNA)
- cell division
what is the function of a cytoskeleton?
- provides shape for cells without cell walls
- movement of cells e.g cillia and flagella
- formation of spindle fibres that move chromosomes
- keep organelles in place
- move organelles e.g vesicles
what is a cytoskeleton?
network of protein fibres
where is the cytoskeleton present?
in eukaryotic cells
what is the plasma membrane around organelles for?
- separate contents from cytoplasm
- controls what can enter and leave the organelle
- surface for reactions to occur on
- eg mitochondria and chloroplasts
what is the plasma membrane around cells for?
- controls what leaves and enters as it is partially permeable
- barriers to certain substances
- keeps contents of cell together
- separates cell contents from the environment
what is the process of protein synthesis?
1) instructions in the nucleus are transcribed into mRNA
2) mRNA leaves nucleus via nuclear pores
3) mRNA travels to ribosomes where message is read and translated into proteins
4) vesicles transport protein to golgi
5) protein fuses to golgi
6) golgi processes and packages protein
7) protein in vesicles are budded off
8) vesicles fuse with [plasma membrane
9) protein is released through membrane pores (exocytosis)
are organelles membrane bound in prokaryotes and eukaryotes?
prokaryotes- non-membrane bound
eukaryotes- both membrane bound and non membrane bound
do prokaryotes and eukaryotes have a nucleus?
prokaryotes- no
eukaryotes- yes
what type of DNA does prokaryotes and eukaryotes have?
prokaryotes- circular
eukaryotes- linear
what DNA organisation do prokaryotes and eukaryotes have?
prokaryotes- proteins fold and condense DNA
eukaryotes- associated with proteins called histones
what is the cell type of prokaryotes and eukaryotes?
prokaryotes- unicellular
eukaryotes- unicellular and multicellular
what are prokaryotes and eukaryotes cell walls made of?
prokaryotes- peptidoglycan
eukaryotes- chitin in fungi
- cellulose in plants
- not present in animals
is the cytoskeleton present in prokaryotes and eukaryotes?
prokaryotes- present
eukaryotes- present but more complex than prokaryotes
do prokaryotes and eukaryotes contain extra chromosomal DNA?
prokaryotes- circular DNA called plasmids
eukaryotes- only present in certain organelles such as chloroplasts and mitochondria
what is diffraction?
the tendency of light waves to spread as they pass close to physical structures
what does metabolism involve?
the synthesis and the breaking down of molecules
what are the three components of the cytoskeleton?
microfilaments, microtubules and intermediate fibres
what are microfilaments?
contractile fibres formed from the protein actin
what are microtubules?
globular tubulin proteins polymerise to form tubes that are used to form a scaffold-like structure that determines the shape of a cell
what are the functions of intermediate fibres?
give mechanical strength to cells and help maintain their integrity
what are centrioles composed of?
microtubules
what is tonoplast?
the membrane of a vacuole in a plant cell
what is safranin dye called in gram staining?
a counterstain
what is resolution limited by?
the diffraction of light
what is the size of the wavelength of the beam of electrons used in electron microscopy?
1nm
what is an artefact and what is it caused by?
a visual structural detail caused by processing the specimen and not a feature of the specimen
what type of microscopes do artefacts appear in?
both light and electron microscopes
what is an example of an artefact?
the bubbles that get trapped under the cover slip
what changes happen to the ultra structure of cells when undergoing prep for electron microscopy?
- loss of continuity in membranes
- distortion in organelles
- empty spaces in the cytoplasm
what is the beam splitter in a laser scanning confocal?
a dichroic mirror, which only reflects one wavelength (from the laser) but allows other wavelengths (produced by the sample) to pass through)
what is the structure of a prokaryote?
a simple structure with just a single undivided internal area
what type of organisms are made up of eukaryotic cells?
animals, plants and fungi
what is the structure of a eukaryote cell?
complicated internal structure, containing many membrane bound nucleus and cytoplasm, which contains many membrane bound cellular components
what type of permeability is a cellulose cell wall and what does it mean?
freely permeable so substances can pass in and out
what do vacuoles contain?
cell sap
what type of permeability is a vacuole and what does it mean?
selectively permeable which means that some small molecules can pass through it but others cannot
what type of permeability is a vacuole that appears in an animal cell?
small and not permeable
when did the first prokaryotic cell appear on earth?
around 3.5 billion years ago
what were the early prokaryotes adapted to?
extremes of salinity, pH and temperature
what are these early prokaryotes called?
extremophiles
where can extremophiles be found today?
in hydrothermal vents and salt lakes
what shape is the one chromosome prokaryotes have and why?
`supercoiled to make it more compact
what are the genes of the chromosome in a prokaryote grouped in to?
operons
what size are ribosomes in prokaryotes and eukaryotes?
prokaryotes- smaller- 70s
eukaryote- larger- 80s
what type of reproduction do prokaryotes and eukaryotes do?
prokaryotes- binary fission
eukaryotes- asexual or sexual
do prokaryotes and eukaryotes have a cell surface membrane?
they both do