Cell structure Flashcards
Define magnification
How many times larger the image is compared to the actual size of the specimen
Define resolution
ability to distinguish between 2 points that are close together
What is the equation for magnification
magnification=image size/real size
Light microscopes
Max magnification:1500x
Max resolution:200nm
-resolution is poor due to long wavelength
-use visible light to form an image
-wavelength of visible light is around 500-650nm
-impossible to distinguish between 2 objects that are closer together than half the wavelength of light
-can view tissues and cells. Eukaryotic cells, their nuclei and possibly mitochondria and chloroplasts
-cannot observe smaller organelles
Advantages of Optical(Light) microscope
-cheap to buy and easy to use
-you don’t need to kill the specimen
disadvantages of optical(light microscope
-lower magnification
-lower resolution
-can’t see the organelles
Electron microscopes
-use electrons to form an image
-a beam of electrons have a smaller wavelength compared to light so it has a higher resolution
-maximum resolution of 0.2nm(can be used to observe small organelles such as ribosomes, endoplasmic reticulum or lysosomes)
-maximum magnification of electron microscope is about 1,500,000
Transmission electron microscopes(TEM)
-electrons are fired at the specimen and pass through the material. It is transmitted through the specimen
-denser parts of the specimen absorb more electrons which makes them appear more darker-produces contrast between different parts of the object being observed
Advantages of TEM
-can see transparent organelles
-high resolution and magnification
-internal structures can be seen
Disadvantages of TEM
-specimen must be dead
-image will be in black and white
-expensive
Scanning electron microscope(SEM)
-electrons are fired at the specimen and the beam bounces off the surface of the material and electrons are detected to create a 3D image
Advantages of SEM
-can be used on thick or 3D specimen
-allows 3D structure of specimen to be observed
Disadvantages of SEM
-specimen must die
-image is in black and white
-expensive
Laser scanning confocal microscope
-thick section of tissue or small living organisms are scanned with a laser beam which is reflected by fluorescent dyes
-computer then assembles an image
advantages of Laser scanning confocal microscope
-specimen can live
-high resolution
-3D image is produced
disadvantages of Laser scanning confocal microscope
-slow process
-laser can cause photodamage to the cells
-very expensive
What is an eyepiece graticule?
-An eyepiece graticule is a disc within the eyepiece lens that has a cm ruler. Full length is always shown and length is fixed
What is a stage micrometre?
It is a glass disc with a measurement
How to calibrate an eyepiece graticule:
-identify a region where the divisions of the eyepiece graticule and stage micrometre line up well
-count divisions of eyepiece graticule and SM in that region
-in 2 columns, write down the eyepiece graticule unit and the length of SM in that region
-divide length of SM by the number of EGU and that gives the length that represents 1 EGU
Magnification
-size of cell is usually measured in micrometres(Um) and cellular structures are usually measured in nanometres or micrometres
-all measurements must be the exact same when doing calculations
magnification=eyepiece lens magnification x objective lens magnification
Resolution
-if 2 separate points cannot be resolved, they will be observed as one point
-resolution of light microscope is limited as wavelength of light is too long
-longer the wavelength of light, the more it is diffracted
-electron microscopes have a higher resolution due to smaller wavelength of beam of electrons compared to light
Light microscope extra
-shine light through specimen which is passed through an objective lens and an eyepiece lens which magnify the specimen
electron microscope extra
-fire a beam of electrons
-shorter wavelength-high resolution
-useful for looking at organelles, viruses and DNA
-requires dead specimen