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