cell structure Flashcards
what is the use of microscopes
the analyse cell components and observe organelles
definition of magnification
how many times bigger the image produced by the microscope is than the real life object you are viewing
definition of resolution
the ability to distinguish between objects that are close together
light microscopes
uses light to form an image
this limits the resolution as it is impossible to distinguish between two objects that are closer than half the wavelength of light (500-650 nm)
what can light microscopes be used for
maximum resolution of 0.2 micrometers
can be used to observe eukaryotic cells, their nuclei and possibly mitochondria and chloroplasts
cannot be used to observe smaller organelles such as ribosomes, endoplasmic reticulum or lysosomes.
maximum useful magnification is about x1500
electron microscopes
uses electrons to form an image
this greatly increases the resolution giving a more detailed image
the beam of electrons has a much smaller wave length than light so an electron microscope can distinguish between two objects extremely close together
what can electron microscopes be used for
maximum resolution of around 0.0002 micrometers
can be used to observe small organelles such as ribosomes, the endoplasmic reticulum or lysosomes
maximum useful magnification is about x1,500,000
transmission electron microscopes
uses electromagnets to focus a beam of electrons which is transmitted through the specimen
denser parts of the specimen absorb more electrons and therefore appear darker on the final image produced
advantages of TEMs
gives high resolution images
allows the internal structures within cells to e seen
disadvantages of TEMs
can only be used on very thin specimens
cannot be used on live specimens ( there is a vacuum inside TEM, all water must be removed so specimen must be dead)
lengthy treatment required to prepare specimens
they do not produce a colour image
scanning electron microscopes
scan electrons across the specimen
beam bounces off the surface of the specimen and the electrons are detected forming a three dimensional image that shows the surface of specimens
advantages of SEM
can be used on thick or 3d specimens
allow the external, 3d structure to be observed
disadvantages of SEM
they give lower resolution images than TEM
they cannot be used to observe live specimens
they do not produce a colour image
laser scanning confocal microscope
relatively new technology
cells being viewed must be stained with fluorescent dyes
a thick section of tissue or small living organisms are scanned with a laser beam, which is reflected by the dyes
multiple depths of the organisms are scanned to produce an image
advantages of laser scanning microscope
can be used on thick or 3d specimens
allows the external 3d structure to be observed
very clear images are produced, high resolution due to the fact the laser beam can be focused at a specific depth
disadvantages of laser scanning microscope
slow process and takes long to obtain an image
the laser has potential to cause photo damage to the cells
different types of sample preparation
dry mount- solid specimens are viewed whole or sectioned into thin slices
wet mount- specimens are suspended in a liquid such as water or immersion oil
squash slides- a wet mount is first prepared then lens tissue is used to gently press down the cover slip, used for soft samples
smear slides- the specimen is smeared onto the slide and a cover slip is placed over
how light microscopes work
light is directed through a thin layer of specimen
light is focused through several lenses so an image can be visible
magnifying power of the microscope can be increased by rotating the objective lens
keep components of a light microscope
the eyepiece lens
objective lens
the stage
the light source
the course and fine focus
Preparing a slide using a liquid specimen
Add a few drops of the sample to the slide using a pipette
Cover the liquid/smear with a coverslip and gently press down to remove air bubbles
Wear gloves to ensure there is no cross-contamination of foreign cells
Methods of preparing a microscope slide using a solid specimen
Take care when using sharp objects and wear gloves to prevent the stain from dying your skin
Use scissors to cut a small sample of the tissue
Peel away or cut a very thin layer of cells from the tissue sample to be placed on the slide (using a scalpel or forceps)
The tissue needs to be thin so that the light from the microscope can pass through
Apply a stain
Gently place a coverslip on top and press down to remove any air bubbles
how can you used chemicals or freezing on specimen samples
Some tissue samples need to be treated with chemicals to kill/make the tissue rigid
This involves fixing the specimen using formaldehyde (preservative), dehydrating it using a series of ethanol solutions, impregnating it in paraffin/resin for support then cutting thin slices from the specimen using a microtome
The paraffin is removed from the slices/specimen, a stain is applied and the specimen is mounted using a resin and a coverslip is applied
OR
Freeze the specimen in carbon dioxide or liquid nitrogen
Cut the specimen into thin slices using a cryostat
Place the specimen on the slide and add a stain
Gently place a coverslip on top and press down to remove any air bubbles