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Lecture 3

BIO1011 Lecture Notes - Lecture 3: Confocal Microscopy, Phase-Contrast Microscopy, Scanning Electron Microscope


Department
Biology
Course Code
BIO1011
Professor
Various
Lecture
3

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Microscopy
There are two key properties of a microscope.
-Magnification - enlarging the image
-Resolution - ability to see fine detail
Types of Microscopes
Light microscopy:
allows us to magnify cells up to 1000 times and to resolve details as small as
0.2um.
Light microscopy allows us to see cells of 400- 700 um.
If a cell is 100um will be unable to see the organelles, due the to the resolution
being unable to work, even if you can keep magnifying.
A bright light must be focused onto the specimen by lenses in the condenser.
The specimen must be carefully prepared to allow light to go through it.
Phase contrast microscopy:
Uses a light microscope with an optical component to take advantage of the
different refractive indexes of light passing through different regions of the
cell.
Bright-field:
Employs a light microscope and requires that samples be fixed and stained
in order to reveal cellular details.
Fluorescence microscopy:
Fluorescent dyes or probes used for staining cells are detected with the aid of a
fluorescence microscope.
This is similar to light microscope except that the illuminating light is passed through
two sets of filters.
Confocal microscopy:
A specialized type of fluorescence microscope that builds up an image by
scanning the specimen with a laser beam.
Electron microscopy:
Rather than using light, electrons are used
There is much greater resolution and magnification than light microscopes.
There is a better level of resolution because the wave lengths involved are shorter
than used in light microscopy.
Electron microscope images are not in colour
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