Variations of Light Microscope

Brightfield Microscope
Brightfield microscopes are the most common
type. A brightfield microscope focuses light
through a condenser lens and passes it through
the specimen. Above the specimen is a tube
containing an objective lens and an eyepiece lens
that together focus the light to form an enlarged
image. Images seen through a brightfield
microscope generally have poor contrast unless
the specimen has been stained with dyes or
chemicals that adhere to certain types of
structures in the cell.



Dark Field Microscopes
Dark field microscopes are similar to brightfield
microscopes but are equipped with a dark field patch
stop and a direct illumination block. Together, these two
components ensure that only light scattered by the
specimen passes through the objective lens. When
viewed through a dark field microscope, a specimen
appears as if it were on a black background. Dark field
microscopy can provide enhanced contrast, although
some of the features that are visible with bright field
microscopy may not be visible with the dark field
technique.



Fluorescence Microscopes
Fluorescent materials absorb light or
electromagnetic radiation of one wavelength and
re-radiate electromagnetic radiation of another
wavelength (typically a longer wavelength). In a
fluorescence microscope, the specimen is first
stained with fluorescent dyes or molecules that
adhere to specific molecules or structures in the
cell. Light from the light source shines on a
dichroic mirror that reflects light of some
wavelengths and transmits light of other
wavelengths. The light from the source is reflected
onto the sample, where the fluorescent dyes
absorb it and emit light that passes through the
dichroic mirror into the objective lens. A fluorescent microscope produces an image in which
only the cell components tagged with fluorescent
dyes are visible.



Confocal Microscope
A confocal microscope is similar to a fluorescence
microscope but incorporates some additional
features, most notably a pinhole aperture that
excludes out-of-focus light. Consequently, the
confocal microscope can form a very sharp image
of a single plane in the sample. Confocal
microscopes are powerful tools but require long
exposure times to form a good image.





Phase Contrast Microscope
The phase contrast microscope is typically used to
examine biological tissue according to NobelPrize.org.
Phase contrast microscopes alter light waves to enhance
the view of a specimen. The phase plate of a phase
contrast microscope increases the wave length of light by
half a wave length. This results in the specimen on the
stage being viewed as a dark object, yet extremely clear
to the eye.

Nomarski Microscopes
Nomarski differential interference contrast
microscopes give specimens a three
dimensional appearance thats unlike the
appearance the specimen would have under
an electron microscope according to Rice
University. Nomarski microscopes are the most
expensive types of light microscopes and
involve passing light through a polarizer
resulting in light vibrating within a single plane.
Light then passes through a small prism which
separates it before passing through the
condenser. Most people try and purchased
used Nomarski microscopes because of their
expensive price.



SIMILARITIES



SEM (Scanning Electron
Microscope)
TEM (Transmission Electron
Microscope)

Both SEM (scanning electron microscope) and TEM (transmission electron
microscope) refer both to the instrument and the method used in electron
microscopy.


Both are types of electron microscopes and give the possibility of seeing,
studying, and examining small, subatomic particles or compositions of a
sample.

Both use electrons (specifically, electron beams), the negative charge of an
atom
Both samples in use are required to be stained or mixed with a particular
element in order to produce images.
Images produced from these instruments are highly magnified and have a high
resolution
DIFFERENCES
SEM (Scanning Electron
Microscope)
TEM (Transmission Electron
Microscope)
Method
Method used is based on scattered
electrons
Method used is based on transmitted
electrons

The scattered electrons in SEM produced
the image of the sample after the
microscope collects and counts the
scattered electrons.




Electrons are directly pointed toward
the sample.
Focus of Analysis
Focuses on the samples surface and its
composition.
Seeks to see what is inside or beyond
the surface

Shows the sample bit by bit Shows the sample as a whole.
Provides a three-dimensional image Delivers a two-dimensional picture.
Magnification and resolution
Only offers 2 million as a maximum level
of magnification


Has up to a 50 million magnification
level
Has 0.4 nanometers Resolution of TEM is 0.5 angstroms
Sample Thickness, Staining and Preparation
Thicker samples


Thinner samples
The sample is prepared on specialized
aluminium stubs and placed on the
bottom of the chamber of the instrument.
Requires the sample to be prepared in
a TEM grid and placed in the middle of
the specialized chamber of the
microscope

The image of the sample is projected
onto the CRT or television-like screen.


The image is produced by the
microscope via fluorescent screens.


References:
http://www.ehow.com/info_8556948_differences-between-sem-tem.html
http://www.differencebetween.net/science/difference-between-tem-and-sem/
http://www.ehow.com/list_5937640_types-light-microscopes.html