Optical microscopy

An optical microscope, in other words a light microscope, uses visible light and lenses to create a magnified image of small objects that could not otherwise be seen with naked eyes. Magnification range of the optical microscope is from 10x to 100x this means that a size of 0.2 micrometer objects can be detected.

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Optical microscopy is typically used in biological research and in material science. Often it is the first step for successful material analysis. It gives a good overview of the materials microstructure and structure-property relationships. Magnification range of the optical microscope is from 10x to 100x. Optical microscopes let us view objects as small as 0.2 micrometres (0.2 thousands of a millimetre or 2 x 10-7 m). For example, cells and large bacterias can be seen. Optical microscopy is often used along with other microscopy techniques. For example, TEM and SEM use electrons to form images from much smaller objects and optical microscopy is used to locate the area of interest from the sample.  There are many variations and methods for optical microscopy. 

Visible light is transmitted through or reflected from the sample. Then the light goes through a single lens or a series of lenses, which leads to a magnified view of the sample. The resulting image can be seen directly by naked eye or it can be imagined. Modern optical microscopes use digital imagining. 

Since biological structures are often colorless, these samples are typically stained to obtain better contrast for the image. Staining process involves use of selective dyes and killing the cells. The sample is also fixed, which means preservation of the sample.

Reflected-light microscopy is used for materials such as metals, ceramics and composites. Variations in the surface arise contrast when the light is reflected from the surface. 

If the sample is transparent, a transmission mode is used. The specimen is usually a tens of micrometres thick slice. Absorption of light differs depending on the different regions and this arises contrast. Method is used for the examination of rocks, polymers and tissue samples.

Polarised light microscopy is a form of transmission mode. Contrast arises from differences in birefringence and thickness of the specimen. Method is used to observe grains, grain orientation and thickness.

The optical microscopy has some drawbacks. The technique can produce images from materials that have enough contrast or from strongly refracting objects. Sample thickness is also restricted in the case of transmission mode to tens of micrometers. The resolution of the optical microscope is limited to 0.2 micrometers and the practical magnification limit is 1000x. For smaller objects SEM, TME and AFM are preferred methods.

Suitable sample matrices

  • Circuit boards
  • Protective coatings and paints
  • Cell and tissue samples
  • Pollen
  • Fibers such as asbestos
  • Minerals and rock

Ideal uses of optical microscopy

  • Optical microscopy is often used in materials research to analyze the object's surface for more detailed further studies.
  • In material science structural studies are typically done with an optical microscope.
  • The method is used to examine mineral and rock samples.
  • An optical microscope is used to study cellular structures, even large bacteria can be observed.

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Frequently asked questions

What is optical microscopy commonly used for?

Optical microscopy is used in material science to analyze structures. It can be used alone or in combination with other microscopy techniques, such as TEM or SEM.

It is used for cell structure imaging. For example, it is a common tool in biology and medical diagnostic.

What are the limitations of optical microscopy?

The resolution of the optical microscope is limited to 0.2 micrometers and the magnification is limited to 1000x.

What kind of samples can be analyzed with optical microscopy?

Almost all kinds of samples are usable if light passes through or reflects from the sample. A Typical method is used to analyze material surfaces or it is used to produce an overall image of cell structures.

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