A microscope is used to magnify the image of tiny objects. The objects are clearly seen with a microscope because at least one lens magnifies the image. This lens refracts the light so that it enters the eye and magnifies distant objects.

where dA is the distance of two infinitesimally small “point” objects that can be just separated, l the wavelength of the illuminating light, and NA the numerical aperture of the objective lens. Obviously, the magnification of the lens has no influence on the optical resolution.

A microscope uses two smaller lenses, i.e., an objective lens near the sample and an ocular lens near the observer. The magnification of both these lenses can be the same or different from one another. Multiplying the magnification of each lens yields the overall magnification of the microscope. With a 10x ocular and a 30x objective, the microscope's total magnification is 300x.

Multiplication of equation 5 by the magnification factor of the lens in use, we get a quantity equivalent to FN, but now taking the actual scanning conditions into account. We call this the resolved field number RFN:

The blank is clamped into a vice and kept in place beneath a diamond-tipped, cylinder-shaped cutter that spins at high speed. The blank's surface is trimmed with this cutter in the desired curvature.

Depthof field microscope

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Fieldnumbermicroscope

Optical glass is often used to create microscopic lenses. It is considerably more uniform and has higher purity than conventional glass.

The field number (FN) for optical microscopes indicates the field of view (FOV). It corresponds to the area in the intermediate image that is observable through the eyepieces. Although, we cannot observe very large fields at once, the human eye can scan and integrate structural features over the full field. Furthermore, it is essential, that field size and resolution fit to the capabilities of the human eye.

If we use a lens that has a magnification of 63x, then the calculated field (equation 1 for FN) would correspond to an area of 5.6 mm x 5.6 mm in the intermediate image plane. The diameter of that square is then only 8 mm and much smaller than typical FNs for which the microscope is designed.

Zhang, Y., & Gross, H. (2017). Systematic Design of Microscopic Lenses. Optical Design and Fabrication 2017 (Freeform, IODC, OFT) (2017), Paper IW4A.1, IW4A.1. https://opg.optica.org/abstract.cfm?uri=IODC-2017-IW4A.1

Ahmed, Usman. 2022. What is the Role of Lenses in Microscopy?. AZoOptics, viewed 25 November 2024, https://www.azooptics.com/Article.aspx?ArticleID=2339.

The calculated FN for a confocal microscope is, therefore, the scanned field size of the sample multiplied by the magnification. The intermediate image does not appear during imaging. By tuning the scanning amplitude, the observed field can be changed from a maximum down to zero (3). Zero amplitude means that a single, fixed spot in the sample is observed, as is required for fluorescence correlation spectroscopy ( FCS ). Intermediate amplitudes generate fields smaller than the maximum, constituting a zoom-in function, because the smaller field is displayed on the same monitor size and, consequently, represents an additional “enlargement” (not an optical magnification).

Furthermore, the surface morphology of materials can be observed with the help of a tunneling microscope. Such microscopes use a beam of electrons that can tunnel through the surface of objects at incredibly small distances and form an image of the surface.

The mixture becomes a very thick liquid and is poured into lens molds at this stage. The annealing is carried out at 500 °C after cooling the mixture to approximately 300 °C. Annealing helps eliminate the internal stresses developed during the early cooling phase and weakens the glass. The glass is then gradually cooled to room temperature, and pieces are removed from the molds. These pieces are called blanks.

After cutting, a lens inspection is carried out, and if the required curvature is not achieved, cutting is done again. This process requires a few minutes to well over an hour.

To preserve the optical resolution when scanning a sample and storing the intensities into a digital image, the size of the picture elements projected back from the sample should be smaller than the optical resolution. Otherwise, the digital image has less information than the optical image. As a rule, we assume a factor of ½ times dA as an appropriate oversampling (special image restoring algorithms usually require even higher oversampling, up to fourfold). The required minimal pixel size (dp) can, therefore, be calculated as:

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Lorenz, K. O., Kakkassery, J., Boree, D., & Pinto, D. (2014). Atomic force microscopy and scanning electron microscopy analysis of daily disposable limbal ring contact lenses. Clinical and Experimental Optometry, 97(5), 411–417. https://www.tandfonline.com/doi/abs/10.1111/cxo

While examining a slide or an object in a microscope, the lens closest to it is called the objective lens, which collects light and increases the magnification of the object being examined.

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A condenser lens is located between the light source and the slide platform. It focuses the beam of light on the object and further passes it to the objective lens. The diaphragm controls the quantity of light entering the condenser lens. Anytime a different objective lens can be used to view the item, the amount of light entering the lens can be changed. With magnifications of 400x or more, condenser lenses are very useful.

Field of view microscope40x

A conventional microscope employs numerous lenses and a light source to significantly enhance the image of the object under examination.

Most compound microscopes use four objective lenses, including a low-power lens, a high-power lens, a scanning lens, and an oil-immersion lens.

Ahmed, Usman. (2022, November 02). What is the Role of Lenses in Microscopy?. AZoOptics. Retrieved on November 25, 2024 from https://www.azooptics.com/Article.aspx?ArticleID=2339.

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For widefield microscopes, the FN is set by the diameter of a metal diaphragm in the eyepiece. Its size is set to ensure good image quality within the observed field, as all optical systems cause increasing optical aberration far from the center of the lens. Nevertheless, the definition of “good quality” is only standardized for the flatness of field (1), leaving room for discussion about other aberrations.

Field of view microscopeformula

Ahmed, Usman. "What is the Role of Lenses in Microscopy?". AZoOptics. 25 November 2024. .

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Ahmed, Usman. "What is the Role of Lenses in Microscopy?". AZoOptics. https://www.azooptics.com/Article.aspx?ArticleID=2339. (accessed November 25, 2024).

The compound microscope uses a series of lenses to magnify the image. These lenses are made of optical glass, which is significantly purer and clearer than regular glass.

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These F-theta lenses by Avantier are designed for consistent spot size and uniform field curvature correction, ideal for high-resolution imaging applications.

The human eye's biological lens is also convex because it focuses light onto the retina, where rod and cone cells are located to enable vision.

There are various kinds of microscopes used for magnification. An optical microscope is the most common type, creating an image from visible light using lenses. Another commonly used type of microscope is an electron microscope which uses an electron beam to form images.

Field of view microscope4x

If, for example, we assume λ = 500 nm and NA = 1.4, we have dp = 90 nm. With a scan format of x = y = 1000 pixels, then the field would cover an area of 90 x 90 µm.

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The field number (FN) in microscopy is defined as the diameter of the area in the intermediate image plane that can be observed through the eyepiece. A field number of, e.g., 20 mm indicates that the observed sample area after magnification by the objective lens is restricted to a diameter of 20 mm. If the objective lens has a magnification of, say, 40x with an FN of 20 mm, a sample area with a diameter of 500 µm can be observed.

These lenses provide magnification of 4x, 10x, 40x, and 100x, respectively. Generally, shorter lenses have less magnification power than longer ones.

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Cameras have a fixed observation field and cannot scan the full FOV of the microscope optics. Consequently, the field must be reduced (by insertion of additional optical components) so that the image can be projected onto the camera sensor without losses.

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Magnesium fluoride is commonly used as an anti-reflective coating on lenses. If a microscope contains a mirror, it is often made up of Pyrex glass. Silica (SiO2) is often used as a protective coating for mirrors, whereas aluminum is used for reflective coatings.

We assume a microscope that is specified with an FN of 22 mm (Fig. 1, left). We use a lens with a magnification of 40x and NA of 1.3, the light wavelength is 500 nm, and we scan a square with x = y = 2496 pixels (considered the maximum the instrument can scan). The RFN then equals 13,6 mm.

Nov 21, 2021 — The total magnification of a microscope is equal to an objective lens magnification multiplied by the eyepiece magnification.

In scanning microscopes, like true confocal laser scanners, the FN indicates the maximum area over which the sample can be scanned, but the actual scan process must be set with the correct parameters or resolution may be lost. This condition usually means smaller fields should be observed or very large image formats used. To be able to compare these restrictions, a modified field number (RFN) is introduced. RFN refers to the maximum resolved field.

To characterize the power of a confocal scanning microscope, it is not appropriate to discuss the FOV as the full area that is optically corrected, but rather the field that is possible to scan without losses of optical resolution. With the largest possible rectangular scan format (x,y)max, we can calculate the resolved field size (dsc res):

The shape of the lens has a considerable effect on the refraction of light. In microscopes, convex lenses are usually employed because of their ability to concentrate light on a specific spot.

The raw materials and specific optical glass are combined in the correct ratios. This cullet (i.e., optical glass) serves as a flux. A flux helps lower the temperature at which raw materials normally react. A glass furnace is typically used for melting this mixture around 1400 °C. The temperature may fluctuate depending on the type of lens being manufactured.

Field of view microscope10X

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In confocal scanning microscopes, an intermediate image as such is actually not generated. In fact, the pinhole is located in the intermediate image plane. The intensity as a function of time that passes the pinhole, codes for the image information. These intensity changes are subsequently used to reconstruct the image by distribution of the appropriate time-intensity segments into a frame store. The frame store is filled synchronously to the position of the focus spot in the sample. The frame has a predefined size of x times y picture elements (pixels). The tuple (x,y) is called the scan-format or sometimes scan-resolution (not to be confused with the optical resolution (2)).

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Microscopic lenses result in higher magnification of the object under examination to the observer. At higher magnification, it becomes easy to analyze even minute details of the object. By using multiple lenses in a microscope, object image becomes clearer and easier to examine. With the help of multiple lenses, an object image can even be magnified more than 1000 times.

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Field of view microscopeCalculator

Usually, the magnification value is engraved on the lens barrel and the field number is engraved on the eyepiece barrel. With that information, you can always calculate the diameter of the FOV you are currently observing:

The optical properties of optical glass depend highly upon its composition, which includes a mixture of boron oxide, sodium oxide, barium oxide, zinc oxide, potassium oxide, or lead oxide.

To drive air bubbles to the surface, the temperature is raised to 1550 °C. The mixture is then steadily cooled to 1000 °C with continuous agitation.

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Largefield of view microscope

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The lens the observer looks through when using a microscope is called an ocular lens. It takes light from the objective lens and re-magnifies it to show a large image. In most cases, the ocular lens magnifies 10x or 15x.

A microscope is an additional lens placed in front of your eye. The microscope lens functions like a magnifying glass, bending light to make the object appear wider to get the desired magnification effect. However, a single large lens provides blurry and dark images.

In a second example, we assume a microscope having an FN of 25 mm (Fig 1, right). The same lens (40x/1.3) and light wavelength (500 nm) are used, but the instrument is limited to a maximum of 1024 pixels per dimension. With equation 6, we calculate the RFN to be 5,6 mm. Although for this example, the FN is nominally somewhat larger than in the previous one, the available scan area is much smaller (if we do not accept losses in resolution).

Usman holds a master's degree in Material Science and Engineering from Xian Jiaotong University, China. He worked on various research projects involving Aerospace Materials, Nanocomposite coatings, Solar Cells, and Nano-technology during his studies. He has been working as a freelance Material Engineering consultant since graduating. He has also published high-quality research papers in international journals with a high impact factor. He enjoys reading books, watching movies, and playing football in his spare time.

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Fowler, S. A., & Allansmith, M. R. (1981). The Effect of Cleaning Soft Contact Lenses: A Scanning Electron Microscopic Study. Archives of Ophthalmology, 99(8), 1382–1386. https://jamanetwork.com/journals/jamaophthalmology/article-abstract/633879