Bright-field microscopy is used extensively in microbiology, biotechnology, mineralogy, microelectronics, nanophysics, and pharmaceutic research.

... Numerická apertura; de Numerische Apertur; en Numerical aperture; es Apertura numérica; fa روزنه عددی; fr Ouverture numérique; he מפתח נומרי; hu Numerikus ...

In a Bright field microscope, the microscopic field is brightly lit while the microorganisms appear dark as they are only able to absorb some of the light. The visualization of microorganisms can be improved by staining them with a dye which greatly improves their light-absorbing ability. As a result, greater contrast and color differentiation is achieved when the dye is used.

Darkfieldmicroscope

It is part of the microscope whose function is to hold the set of objective lenses and also allow switching between objective lenses

Resolution in light microscopy can be maximized by the use of the shortest wavelength of visible light and an objective with the maximum NA.

Bright field microscopyadvantages and disadvantages

The resolving power of the microscope is based on the wavelength of light used and the numerical aperture (NA) of the lens.

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It includes other features such as a diaphragm and/or filters which manages the quality and intensity of the illumination.

The largest magnification of the microscope may not be the best for visualization as the images might be fuzzy i.e. greater magnification without greater resolution does not provide benefits.

Bright field microscopyand darkfield microscopy

If magnification is more than the resolving value, the produced larger images will be fuzzy with less distinction in details.

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Numerous sources of light can be used for Bright field microscopy – the simplest being daylight which is directed via a mirror

In order to center the specimen, the focus is done at lower magnification and if higher magnification is required, the stage is moved vertically up and re-focused at higher magnification.

Bright field microscopyprinciple

The limit of resolution is the smallest distance at which two objects can be placed separately and still be distinguishable as two separate objects.

Since moving the slide by hand at 100× magnification is not practical, the stage has control knobs. These control knobs allow tiny horizontal movements of the slide.

Resolving powers the ability to distinguish two adjacent points as distinct and separate. In other words, the ability to distinctly see separate lines or dots and the greater the ability is, the higher the resolution power of a microscope.

The numerical aperture (NA) is the measure of the aperture of the objective (half-aperture angle) – the angle θ subtended by the optical axis and the outermost rays still covered by the objective.

Darkfield microscopy

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Bright fieldmicroscope application

Condenser Diaphragm- This diaphragm controls the amount of light entering the lens system. This feature is useful for viewing unstained biological specimens ...

The interchangeable eyepieces, which can be inserted with different degrees of magnification, are inserted into the top end of the body tube.

extent and magnitude of the diffraction patterns are affected by the wavelength of light used, refractive materials used, and the numerical aperture (NA).

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Bright fieldmicroscope image

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Stage, which is able to move up and down, has a hole at the center through which light passes to illuminate the specimen.

In high-performance microscopes (commonly equipped with apochromatic objectives), the optical configuration of the objective lens and eyepiece are matched to give the best possible optical visualization.

Advantages ofbright-fieldmicroscope

If the objective lenses have higher magnifications, a higher numerical aperture is present, which results in a shorter depth of field.

The objective lens has two parameters – magnification (5× to 100×) and numerical aperture (0.14 to 0.7) – with focal lengths of about 40 to 2 mm respectively.

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Typically, three objective lenses are screwed into a circular nose piece which can be rotated to select the one with a desired objective lens.

Limitation includes the degree to which microscope objectives can be altered to increase and the wavelength of light used – between 400nm (blue light) and 700nm (red light).

Generally, a Bright field microscope is able to produce a magnification of about X1,000 to X2,000. If the object is magnified over the range, images become fuzzy.

Bright-field microscopy is a type of optical microscope or light microscope that uses a system of lenses and visible light to generate magnified images.

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