There are also long-period fiber gratings that have index modulations with periods of hundreds of microns (see Laser Focus World, June 1996, p. 293). Instead of producing a reflected signal, these gratings create a phase-matching, or Bragg, condition that couples a forward-traveling signal into forward-traveling cladding modes. The signals coupled into the cladding are absorbed by the coating, creating a loss. Long-period gratings thus act as wavelength-selective absorption filters and are used in wavelength-division-multiplexing networks and in gain-shaping filters for rare-earth-doped fiber amplifiers.

Microscopeparts and functions

Light incident on a diffraction grating is dispersed away from the grating surface at an angle dependent on its wavelength, allowing a grating to be used to select a narrow spectral band from a much wider band. This ability of a grating is particularly useful for laser tuning, especially in the visible region of the spectrum. Two primary configurations for selecting a narrow wavelength are Littrow and Littman. In the Littrow configuration, the wavelength of interest diffracts at exactly the same angle as the light incident on the grating. Littrow tuning is done either with fine-pitch first-order gratings (typically 1800 or 2400 grooves/mm, either ruled or holographic) or a coarser grating used in higher orders. The alternative approach is to use the grating in a fixed grazing incidence mode together with a rotating reflecting mirror.

"The grooves are similar to the indentations made by a plow in soil," says John Hoose of Richardson Grating Laboratory (Rochester, NY), except that they are much closer together. Anywhere from one to 10,000 fine parallel lines per millimeter can be engraved. Light waves diffracted from these lines interfere, and all wavelengths but one are canceled in any particular direction through destructive interference. The depth of the groove changes the wavelength of the light wave being diffracted.

An advantage of a transmission volume grating is its relative insensitivity to angle, says James Arns of KOSI. A Bragg-type structure follows the classical grating equation concerning image position but with the added ability to adjust the intensity profile over a range of wavelengths. To describe the capability, Arns compares a Venetian blind to lines painted on a window. When the blind is positioned with the slats horizontal, it diffracts light in the same way as the painted lines or a surface-relief grating. When the slats are angled, the element of depth is added to how the light is diffracted. Because of this added dimension, the grating efficiency can be adjusted over the wavelength bandwidth to favor one side or the other. Also, the low sensitivity to incidence angle means the grating can be angularly tuned without influencing the image position.

Coarse adjustmentmicroscope function

Joseph Fraunhofer first used diffraction gratings in 1819 to observe the spectrum of the sun. Earliest devices were multiple-slit assemblies, consisting of a grid of fine wire or thread wound about and extending between two parallel screws, which served as spacers. A wavefront that passed through the system was confronted by alternate opaque and transparent regions, so that it underwent a modulation in amplitude.

Although today’s microscopes are usually far more powerful than the microscopes used historically, they are used for much the same purpose: viewing objects that would otherwise be indiscernible to the human eye.  Here we’ll start with a basic compound microscope and go on to explore the components and function of larger more complex microscopes. We’ll also take an in-depth look at one of the key parts of a microscope, the objective lens.

Since the invention of the replication technique, diffraction gratings have replaced prisms in many commercial spectrometers. A prism will bend short wavelengths more than longer ones (see Laser Focus World, Jan. 1997, p. 101). Prisms that transmit visible light absorb most UV and infrared wavelengths, whereas reflection gratings can be suitably coated for high reflectivity in wide spectral regions. Gratings are considered superior to prisms in many applications. Seeking to combine the best of both, Richardson Grating Laboratory has fabricated a "grism," a part-grating, part-prism optical element useful in spectrometers that require in-line presentation of the spectrum, as in astronomy. The light diffracted by the grating is bent back in line by the refracting effect of the prism. The dispersion of the grism is not linear, because the dispersive effects of the prism and grating are superimposed.

There are two major specifications for a microscope: the magnification power and the resolution. The magnification tells us how much larger the image is made to appear. The resolution tells us how far away two points must be to  be distinguishable. The smaller the resolution, the larger the resolving power of the microscope. The highest resolution you can get with a light microscope is 0.2 microns (0.2 microns), but this depends on the quality of both the objective and eyepiece.

There are some important specifications and terminology you’ll want to be aware of when designing a microscope or ordering microscope objectives. Here is a list of key terminology.

Most microscopes rely on background illumination such as daylight or a lightbulb rather than a dedicated light source. In brightfield illumination (also known as Koehler illumination), two convex lenses, a collector lens and a condenser lens,  are placed so as to saturate the specimen with external light admitted into the microscope from behind. This provides a bright, even, steady light throughout the system.

At Avantier we produce high quality microscope objectives lenses, ocular lenses, and other imaging systems. We are also able to provide custom designed optical lenses as needed. Chromatic focus shift, working distance, image quality, lens mount, field of view, and antireflective coatings are just a few of the parameters we can work with to create an ideal objective for your application. Contact us today to learn more about how we can help you meet your goals.

Pairs of diffraction gratings can also be used to compress or stretch a laser pulse. When a spectrally broad laser pulse is incident on a diffraction grating, the various wavelengths that make up the pulse will diffract from the grating at angles determined by those wavelengths. If the pulse is chirped so that the frequency changes linearly during the length of the pulse, then diffraction will spread the pulse out across the second grating. When the light diffracts from the second grating, which is oriented parallel to the first grating, the different parts of the pulse will diffract at angles that yield a pulse whose parts are synchronized. This increases the peak power while the total energy remains the same. Pulse compression uses two gratings with the same groove frequency and efficiencies peaked for the polarization and wavelength of the laser.

Revolving nosepiecemicroscope function

The parfocal length of a microscope is defined as the distance between the object being studied and the objective mounting plane.

Refractive objectives are so-called because the elements bend or refract light as it passes through the system. They are well suited to machine vision applications, as they can provide high resolution imaging of very small objects or ultra fine details. Each element within a refractive element is typically coated with an anti-reflective coating.

If the gratings are arranged in a nonparallel arrangement, a pulse can be stretched. Pulse stretching uses two identical gratings, allowing lower peak power to be transmitted through the laser system and increasing the amount of stored energy that can be extracted.

Fiber Bragg gratings, another recent development in grating applications, are made within a fiberoptic cable. Fiber gratings are fabricated by exposing the core of a single-mode fiber, 8 to 10 µm thick, to a periodic pattern of intense ultraviolet light. This pattern is created when a 248- or 193-nm laser passes through a special diffractive phase mask. When a fiber is placed in the intense UV light pattern of the mask, a permanent modulation of the index of refraction is generated in the fiber core. This photo-generated index modulation acts as a grating.

Commercial surface-relief gratings are produced using an epoxy casting replication process developed in the mid-1900s. The process involves pouring a liquid into a mold, allowing the liquid to harden, and then removing the hardened material from the mold without damaging either. The replication process yields a grating that is an optically identical copy of the original. The two basic types of grating masters are ruled and interference.

Holoplexing, a technique devised by KOSI in which two gratings are placed together in the same structure to cover multiple spectral ranges at one time, is useful for imaging on charge-coupled-device (CCD) cameras for broadband applications. Holographic transmission gratings are also used in Raman spectroscopy and for pulse compression in ultrafast lasers.

Body tubemicroscope function

A microscope is an optical device designed to magnify the image of an object, enabling details indiscernible to the human eye to be differentiated. A microscope may project the image onto the human eye or onto a camera or video device.

Holographic gratings can also be made from computer-generated interference patterns. The patterns are written onto a chrome mask using an electron-beam machine. The patterns on the mask are then etched into a material, such as fused silica, using photolithographic masking and etching techniques. "Computer-generated gratings have really just reached maturity within the last two years," says Michael Feldman, of Digital Optics Corp. (Charlotte, NC). "They are very flexible and easy to mass-produce."

The eyepiece or ocular lens is the part of the microscope closest to your eye when you bend over to look at a specimen. An eyepiece usually consists of two lenses: a field lens and an eye lens. If a larger field of view is required, a more complex eyepiece  that increases the field of view can be used instead.

The concept of diffraction gratings is simple, yet elegant. For more than one hundred years, they have been used in dispersive optical systems. Applications for gratings are expanding as the fabrication technology grows. Fields as diverse as telecommunications, astronomy, microlithography, lasers, and metal analysis are driving these changes.

A basic achromatic objective is a refractive objective that consists of just an achromatic lens and a meniscus lens, mounted within appropriate housing. The design is meant to limit the effects of chromatic and spherical aberration  as they bring two wavelengths of light to focus in the same plane. Plan Apochromat objectives can be much more complex with up to fifteen elements. They can be quite expensive, as would be expected from their complexity.

A reflective objective works by reflecting light rather than bending it. Primary and secondary mirror systems both magnify and relay the image of the object being studied. While reflective objectives are not as widely used as refractive objectives, they offer many benefits. They can work deeper in the UV or IR spectral regions, and they are not plagued with the same aberrations as refractive objectives. As a result, they tend to offer better resolving power.

Stagemicroscope function

A basic compound microscope could consist of just two elements acting in relay, the objective and the eyepiece. The objective relays a real image to the eyepiece, while magnifying that image anywhere from 4-100x.  The eyepiece magnifies the real image received typically by another 10x, and conveys a virtual image to the sensor.

In modern microscopes, neither the eyepiece nor the microscope objective is a simple lens. Instead, a combination of carefully chosen optical components work together to create a high quality magnified image. A basic compound microscope can magnify up to about 1000x. If you need higher magnification, you may wish to use an electron microscope, which can magnify up to a million times.

The field of view (FOV) of a microscope is simply the area of the object that can be imaged at any given time. For an infinity-corrected objective, this will be determined by the objective magnification and focal length of the tube lens. Where a camera is used the FOV  also depends on sensor size.

The optical performance of an objective is dependent largely on the optical aberration correction, and these corrections are also central to image quality and measurement accuracy. Objective lenses are classified as achromat, plan achromat, plan semi apochromat, plan apochromat, and super apochromat depending on the degree of correction.

Grating applicationsLight incident on a diffraction grating is dispersed away from the grating surface at an angle dependent on its wavelength, allowing a grating to be used to select a narrow spectral band from a much wider band. This ability of a grating is particularly useful for laser tuning, especially in the visible region of the spectrum. Two primary configurations for selecting a narrow wavelength are Littrow and Littman. In the Littrow configuration, the wavelength of interest diffracts at exactly the same angle as the light incident on the grating. Littrow tuning is done either with fine-pitch first-order gratings (typically 1800 or 2400 grooves/mm, either ruled or holographic) or a coarser grating used in higher orders. The alternative approach is to use the grating in a fixed grazing incidence mode together with a rotating reflecting mirror. Pairs of diffraction gratings can also be used to compress or stretch a laser pulse. When a spectrally broad laser pulse is incident on a diffraction grating, the various wavelengths that make up the pulse will diffract from the grating at angles determined by those wavelengths. If the pulse is chirped so that the frequency changes linearly during the length of the pulse, then diffraction will spread the pulse out across the second grating. When the light diffracts from the second grating, which is oriented parallel to the first grating, the different parts of the pulse will diffract at angles that yield a pulse whose parts are synchronized. This increases the peak power while the total energy remains the same. Pulse compression uses two gratings with the same groove frequency and efficiencies peaked for the polarization and wavelength of the laser. If the gratings are arranged in a nonparallel arrangement, a pulse can be stretched. Pulse stretching uses two identical gratings, allowing lower peak power to be transmitted through the laser system and increasing the amount of stored energy that can be extracted. Since the invention of the replication technique, diffraction gratings have replaced prisms in many commercial spectrometers. A prism will bend short wavelengths more than longer ones (see Laser Focus World, Jan. 1997, p. 101). Prisms that transmit visible light absorb most UV and infrared wavelengths, whereas reflection gratings can be suitably coated for high reflectivity in wide spectral regions. Gratings are considered superior to prisms in many applications. Seeking to combine the best of both, Richardson Grating Laboratory has fabricated a "grism," a part-grating, part-prism optical element useful in spectrometers that require in-line presentation of the spectrum, as in astronomy. The light diffracted by the grating is bent back in line by the refracting effect of the prism. The dispersion of the grism is not linear, because the dispersive effects of the prism and grating are superimposed.New fabrication techniquesKaiser Optical Systems Inc. (KOSI; Ann Arbor, MI), has developed an alternative to the classical or surface-relief holographic grating--the volume transmission holographic grating (see photo at top of this page; also Laser Focus World, Oct. 1995, p. 95). The grating is created in the traditional manner by recording interference patterns generated by two mutually coherent laser beams. After the pattern is defined in the photosensitive material, coated on glass, and the film developed, a top layer of glass is added, creating a totally transparent grating assembly. Light strikes the grating on one side and diffracts out through the other.An advantage of a transmission volume grating is its relative insensitivity to angle, says James Arns of KOSI. A Bragg-type structure follows the classical grating equation concerning image position but with the added ability to adjust the intensity profile over a range of wavelengths. To describe the capability, Arns compares a Venetian blind to lines painted on a window. When the blind is positioned with the slats horizontal, it diffracts light in the same way as the painted lines or a surface-relief grating. When the slats are angled, the element of depth is added to how the light is diffracted. Because of this added dimension, the grating efficiency can be adjusted over the wavelength bandwidth to favor one side or the other. Also, the low sensitivity to incidence angle means the grating can be angularly tuned without influencing the image position."It also has a high efficiency," says Arns. "Depending on the configuration, the grating can produce 90% efficiency in the first order. If the thickness or the frequency of the grating is high enough, higher orders that otherwise might be propagated are extinguished." Another advantage, says Arns, is that the element can be handled and cleaned in the same fashion as a high-quality cemented lens because the grating is sandwiched between two layers of glass. Also, because the Bragg-type grating is a transmission device, optical elements and instruments can be brought close to it, resulting in a compact design.Holoplexing, a technique devised by KOSI in which two gratings are placed together in the same structure to cover multiple spectral ranges at one time, is useful for imaging on charge-coupled-device (CCD) cameras for broadband applications. Holographic transmission gratings are also used in Raman spectroscopy and for pulse compression in ultrafast lasers.Holographic gratings can also be made from computer-generated interference patterns. The patterns are written onto a chrome mask using an electron-beam machine. The patterns on the mask are then etched into a material, such as fused silica, using photolithographic masking and etching techniques. "Computer-generated gratings have really just reached maturity within the last two years," says Michael Feldman, of Digital Optics Corp. (Charlotte, NC). "They are very flexible and easy to mass-produce." Their versatility offers many advantages. "Ruled and holographic gratings are limited to relatively simple structures by the fabrication methods that are used," says W. Hudson Welch, also of Digital Optics. "The flexibility provided by computer-generated gratings allows the creation of essentially arbitrary grating patterns."Fiber gratingsFiber Bragg gratings, another recent development in grating applications, are made within a fiberoptic cable. Fiber gratings are fabricated by exposing the core of a single-mode fiber, 8 to 10 µm thick, to a periodic pattern of intense ultraviolet light. This pattern is created when a 248- or 193-nm laser passes through a special diffractive phase mask. When a fiber is placed in the intense UV light pattern of the mask, a permanent modulation of the index of refraction is generated in the fiber core. This photo-generated index modulation acts as a grating. Light traveling along the fiber core impinges on the grating, and each area of different refractive index scatters a small portion of the beam. If the wavelength of the signal is twice the distance between the periodic refractive elements (typically <1 µm), then the signals scattered back down the fiber core will add constructively to give a large reflection. The wavelength at which the reflection occurs is the Bragg wavelength. A Bragg grating can operate at precise wavelengths that can be accurately preset and maintained, says Keith Brundin at 3M Specialty Optical Fibers (West Haven, CT).There are also long-period fiber gratings that have index modulations with periods of hundreds of microns (see Laser Focus World, June 1996, p. 293). Instead of producing a reflected signal, these gratings create a phase-matching, or Bragg, condition that couples a forward-traveling signal into forward-traveling cladding modes. The signals coupled into the cladding are absorbed by the coating, creating a loss. Long-period gratings thus act as wavelength-selective absorption filters and are used in wavelength-division-multiplexing networks and in gain-shaping filters for rare-earth-doped fiber amplifiers. Fiber Bragg gratings have been commercially available only since 1995. They are becoming increasingly popular in telecommunications and the laser industry for such applications as external reflectors for stabilizing semiconductor lasers (see Fig. 4) and single- frequency fiber lasers.

Historically microscopes were simple devices composed of two elements. Like a magnifying glass today, they produced a larger image of an object placed within the field of view. Today, microscopes are usually complex assemblies that include an array of lenses, filters, polarizers, and beamsplitters. Illumination is arranged to provide enough light for a clear image, and sensors are used to ‘see’ the object.

Light traveling along the fiber core impinges on the grating, and each area of different refractive index scatters a small portion of the beam. If the wavelength of the signal is twice the distance between the periodic refractive elements (typically <1 µm), then the signals scattered back down the fiber core will add constructively to give a large reflection. The wavelength at which the reflection occurs is the Bragg wavelength. A Bragg grating can operate at precise wavelengths that can be accurately preset and maintained, says Keith Brundin at 3M Specialty Optical Fibers (West Haven, CT).

Their versatility offers many advantages. "Ruled and holographic gratings are limited to relatively simple structures by the fabrication methods that are used," says W. Hudson Welch, also of Digital Optics. "The flexibility provided by computer-generated gratings allows the creation of essentially arbitrary grating patterns."

Armmicroscope function

"It also has a high efficiency," says Arns. "Depending on the configuration, the grating can produce 90% efficiency in the first order. If the thickness or the frequency of the grating is high enough, higher orders that otherwise might be propagated are extinguished." Another advantage, says Arns, is that the element can be handled and cleaned in the same fashion as a high-quality cemented lens because the grating is sandwiched between two layers of glass. Also, because the Bragg-type grating is a transmission device, optical elements and instruments can be brought close to it, resulting in a compact design.

The working distance of a microscope is defined as the free distance between the objective lens and the object being studied. Low magnification objective lenses have a long working distance.

Both the objective lens and the eyepiece also contribute to the overall magnification of the system. If an objective lens magnifies the object by 10x and the eyepiece by 2x, the microscope will magnify the object by 20. If the microscope lens magnifies the object by 10x and the eyepiece by 10x, the microscope will magnify the object by 100x. This multiplicative relationship is the key to the power of microscopes, and the prime reason they perform so much better than simply magnifying glasses.

Diffraction gratings are fundamental optical elements that have a precise pattern of grooves superimposed on them. These minute, periodic structures diffract, or disperse, incident light in such a way that the individual wavelengths making up the incident light can be differentiated. Gratings are indispensable in helping physicists determine the structure of atoms or helping astronomers calculate the chemical composition of stars and the rotation of galaxies. Applications are expanding; one of the fastest growing areas for gratings—laser pulse compression—didn’t even exist until a few years ago.

Numerical aperture NA denotes the light acceptance angle. Where θ is the maximum 1/2 acceptance ray angle of the objective and n is the index of refraction of the immersive medium, the NA can be denoted by

Basemicroscope function

Microscope objective lenses are typically the most complex part of a microscope.  Most microscopes will have three or four objectives lenses, mounted on a turntable for ease of use. A scanning objective lens will provide 4x magnification,  a low power magnification lens will provide magnification of 10x, and a high power objective offers 40x magnification. For high magnification, you will need to use oil immersion objectives. These can provide up to 50x, 60x, or 100x magnification and increase the resolving power of the microscope, but they cannot be used on live specimens.

Kaiser Optical Systems Inc. (KOSI; Ann Arbor, MI), has developed an alternative to the classical or surface-relief holographic grating--the volume transmission holographic grating (see photo at top of this page; also Laser Focus World, Oct. 1995, p. 95). The grating is created in the traditional manner by recording interference patterns generated by two mutually coherent laser beams. After the pattern is defined in the photosensitive material, coated on glass, and the film developed, a top layer of glass is added, creating a totally transparent grating assembly. Light strikes the grating on one side and diffracts out through the other.

While a magnifying glass consists of just one lens element and can magnify any element placed within its focal length, a compound lens, by definition, contains multiple lens elements. A relay lens system is used to convey the image of the object to the eye or, in some cases, to camera and video sensors.

An microscope objective  may be either reflective or refractive. It may also be either finite conjugate or infinite conjugate.

Nosepiecemicroscope function

The author wishes to thank John Hoose of Richardson Grating Laboratory (Rochester, NY) for his help in preparing this article.

While most microscope objectives are designed to work with air between the objective and cover glass, objectives lenses designed for higher NA and greater magnification sometimes use an alternate immersion medium. For instance, a typical oil immersion object is meant to be used with an oil with refractive index of 1.51.

In 1882, Henry A. Rowland invented the process of ruling, or scratching parallel notches into metal deposited onto the surface of a flat, clear glass plate—a method that produced gratings of exceptionally high quality. Modern ruled gratings can be either reflective or transmissive and are fabricated with a single diamond point that burnishes grooves on flat or concave surfaces.