There are also several techniques that can be implemented to surpass the resolution limit of the transmitted light. For example, holographic techniques have been shown to reach a higher resolution in one experimental study. Other methods to increase the resolution of an optical microscope include:

Scanning tunnelingmicroscope

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Smith, Yolanda. (2018, August 23). Limitations of Optical Microscopy. News-Medical. Retrieved on November 03, 2024 from https://www.news-medical.net/life-sciences/Limitations-of-Optical-Microscopy.aspx.

202422 — The focal length of a lens is the optical distance from the point where the light meets inside the lens to the camera's sensor.

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Optical microscopy is a very useful technique to examine the appearance of a sample with greater detail, but there are some limitations that provide a boundary to its use in practice.

The magnitude of diffraction and, hence, the resolving power of an optical microscope depends on the light wavelength (λ) and the numerical aperture (NA) of the objective lens.  As a result, there is a distinct limit of an optical microscope to view adjacent structural details clearly, which is known as the diffraction limit of the microscope.

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These diffraction rings limit the ability of the optical microscope to resolve fine details of the sample. The resolving power of an optical microscope is a measure of the ability of the microscope to distinguish between two adjacent structural details, without the interference of Airy discs.

The maximum magnification that can be achieved by an optical microscope typically ranges from 500x to 1500x. While this level of magnification has many purposes and can be useful for a number of practical applications, it is considerably lower than the magnification that can be achieved with electron microscopy. In contrast, an electron microscope may be able to provide magnifications greater than 160,000x.

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When an optical microscope with transmitted light is used at very high magnifications, the image of point objects may be distorted. They may be seen as fuzzy discs that are surrounded by diffraction rings, known as Airy discs.

Opticalmicroscope

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Depth of focus is a lens optics concept that measures the tolerance of placement of the image plane (the film plane in a camera) in relation to the lens.

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Additionally, with the use of a fluorescent sample, other techniques can also be used to reach a higher resolution. For example, near-field scanning optical microscopy with evanescent waves and stimulated emission depletion may prove useful.

Type ofmicroscope

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Yolanda graduated with a Bachelor of Pharmacy at the University of South Australia and has experience working in both Australia and Italy. She is passionate about how medicine, diet and lifestyle affect our health and enjoys helping people understand this. In her spare time she loves to explore the world and learn about new cultures and languages.

As a result, the low magnification of an optical microscope is a limiting factor for some applications, when an electron microscope may be better suited to the purpose at hand.

Smith, Yolanda. "Limitations of Optical Microscopy". News-Medical. 03 November 2024. .

Smith, Yolanda. "Limitations of Optical Microscopy". News-Medical. https://www.news-medical.net/life-sciences/Limitations-of-Optical-Microscopy.aspx. (accessed November 03, 2024).

Invertedmicroscope

Smith, Yolanda. 2018. Limitations of Optical Microscopy. News-Medical, viewed 03 November 2024, https://www.news-medical.net/life-sciences/Limitations-of-Optical-Microscopy.aspx.

The primary limitations of optical microscopes, including resolution, magnification and surface view, are described and discussed below. Additionally, some methods to overcome these limitations or alternative types of microscope that may be used instead are covered.

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Similar to the magnification limitation of optical microscopy, the surface view of the sample with an optical microscope is sufficient for many purposes, but can be a limiting factor. It is significantly less clear than what can be achieved with an electron microscope, and this alternative may be preferred in some situations.

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This does not limit its use for many applications though i.e. when high resolution with a clear surface view is not necessary. However, in some instances electron or another type of microscopy may be better suited to the purpose to achieve and maintain a high quality surface view at higher resolutions.

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