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)

Method Method used is based on scattered electrons The scattered electrons in SEM produced the image of the sample after the microscope collects and counts the scattered electrons.

TEM (Transmission Electron Microscope)

Method used is based on transmitted 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 is produced by the microscope via fluorescent screens.

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

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