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Light Microscope: E.G. During Dissection

The light microscope has a maximum magnification of 1500x and resolution of 200nm, allowing living samples to be viewed relatively inexpensively. Compound microscopes magnify hundreds of times while dissecting microscopes provide low magnification with distance for dissection. Samples often require staining before viewing. Electron microscopes use electrons rather than light, achieving 0.5nm resolution at 500,000x magnification. Transmission electron microscopes produce 2D images of thin samples while scanning electron microscopes make 3D surface images. Electron microscope sample preparation is complex and requires skills like fixation, dehydration, embedding, sectioning, and staining.

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154 views2 pages

Light Microscope: E.G. During Dissection

The light microscope has a maximum magnification of 1500x and resolution of 200nm, allowing living samples to be viewed relatively inexpensively. Compound microscopes magnify hundreds of times while dissecting microscopes provide low magnification with distance for dissection. Samples often require staining before viewing. Electron microscopes use electrons rather than light, achieving 0.5nm resolution at 500,000x magnification. Transmission electron microscopes produce 2D images of thin samples while scanning electron microscopes make 3D surface images. Electron microscope sample preparation is complex and requires skills like fixation, dehydration, embedding, sectioning, and staining.

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Dr.Charin
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© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Light microscope

 The maximum magnification of light microscopes is usually ×1500, and their maximum


resolution is 200nm, due to the wavelength of light. An advantage of the light microscope is that it
can be used to view a variety of samples, including whole living organisms or sections of larger
plants and animals. It is also relatively inexpensive.

 There are two types of light microscope. Compound Microscopes contain several lenses and
magnify a sample several hundred times. Dissecting Microscopes on the other hand have a low final
magnification but are useful when a large working distance between the objectives and the stage is
required (e.g. during dissection). They have two eyepieces to produce a 3D stereoscopic view.

 Many specimens require preperation before being viewed by a light microscope, as some


may not be coloured or might distort when cut. Samples are Stained with coloured stains that bind
to certain chemicals or cell structures. For example, Acetic Orcein stains DNA dark red .

Electron Microscope
 Light microscopes low magnification and resolution are
insatisfactory for viewing very small things, like Organelles within
cells. In these circumstances, and Electron Microscope may be used.
Electorns have a much lower wavelength than light (100000 times
shorter in fact, at 0.004nm) which means that they can be used to
produce an image with resolution as great as 0.5nm. Electron
Microscopes can have magnifications of ×500000.

 There are different types of Electron Microscope.


A Transmission Electron Microscope (TEM) produces a 2D image of
a thin sample, and has a maximum resolution of ×500000.
 A Scanning Electron Microscope (SEM) produces a 3D image of a sample by 'bouncing'
electons off and dectecting them at multiple detectors. It has a maximum magnification of
about ×100000.

 The preparation of a sample for electron microscopy is a complex process. It may involve
o Chemical Fixation: Stabilising an
organism/sample's mobile macrostructure
o Cryofixation: Freezing the sample very
rapidly to preserve its state
o Dehydration: Removing the water form a
specimen, for example, by replacing it with ethanol
o Embedding: Embedding in resin, ready
to be sectioned
o Sectioning: Cutting the sample into thin
strips that are semitransparent to electrons, for
example with a diamond knife
o Staining: Using heavy metals to scatter
electrons and produce contrast
o Freeze Fracturing: Freezing the sample
rapidly, and then fracturing it, for example, when
viewing cell membranes
o Mounting: Placing the sample on a copper grid

 It is advantageous to use an Electron Microscope in many situations because they offer


a much higher resolution that Light Microscopes, so they can be used to image very small objects in
detail, and also because of the 3D images that SEMs offer. However, samples must be placed in a
vacuum as electrons are deflected by particles in the air, they are very expensive to buy and
maintain, and preparing the samples requires a lot of skill to do.

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