Optical microscope

A modern optical microscope with a mercury bulb for fluorescence microscopy. The microscope has a digital camera which is connected to a computer.

The optical microscope, also referred to as a light microscope, is a type of microscope that commonly uses visible light and a system of lenses to generate magnified images of small objects. Optical microscopes are the oldest design of microscope and were possibly invented in their present compound form in the 17th century. Basic optical microscopes can be very simple, although many complex designs aim to improve resolution and sample contrast.

The object is placed on a stage and may be directly viewed through one or two eyepieces on the microscope. In high-power microscopes, both eyepieces typically show the same image, but with a stereo microscope, slightly different images are used to create a 3-D effect. The image can also be captured by normal, photosensitive cameras to generate a micrograph. Originally, images were captured by photographic film, but modern developments in CMOS and charge-coupled device (CCD) cameras allow the capture of digital images. Purely digital microscopes are now available which use a CCD camera to examine a sample, showing the resulting image directly on a computer screen without the need for eyepieces.

A range of objective lenses with different magnification are usually provided mounted on a turret, allowing them to be rotated into place and providing an ability to zoom-in. The maximum magnification power of optical microscopes is typically limited to around 1000x because of the limited resolving power of visible light. The magnification of a compound optical microscope is the product of the magnification of the eyepiece (say 10x) and the objective lens (say 100x), to give a total magnification of 1,000×. Modified environments such as the use of oil or ultraviolet light can increase the magnification.

Alternatives to optical microscopy which do not use visible light include scanning electron microscopy and transmission electron microscopy and scanning probe microscopy and as a result, can achieve much greater magnifications.

Types

Diagram of a simple microscope

There are two basic types of optical microscopes: simple microscopes and compound microscopes. A simple microscope uses the optical power of single lens or group of lenses for magnification. A compound microscope uses a system of lenses (one set enlarging the image produced by another) to achieve much higher magnification of an object. The vast majority of modern research microscopes are compound microscopes while some cheaper commercial digital microscopes are simple single lens microscopes. Compound microscopes can be further divided into a variety of other types of microscopes which differ in their optical configurations, cost, and intended purposes.

Simple microscope

A simple microscope uses a lens or set of lenses to enlarge an object through angular magnification alone, giving the viewer an erect enlarged virtual image.[1][2] The use of a single convex lens or groups of lenses are found in simple magnification devices such as the magnifying glass, loupes, and eyepieces for telescopes and microscopes.

Compound microscope

Diagram of a compound microscope

A compound microscope uses a lens close to the object being viewed to collect light (called the objective lens) which focuses a real image of the object inside the microscope (image 1). That image is then magnified by a second lens or group of lenses (called the eyepiece) that gives the viewer an enlarged inverted virtual image of the object (image 2).[3] The use of a compound objective/eyepiece combination allows for much higher magnification. Common compound microscopes often feature exchangeable objective lenses, allowing the user to quickly adjust the magnification.[3] A compound microscope also enables more advanced illumination setups, such as phase contrast.

Other microscope variants

There are many variants of the compound optical microscope design for specialized purposes. Some of these are physical design differences allowing specialization for certain purposes:

  • Stereo microscope, a low-powered microscope which provides a stereoscopic view of the sample, commonly used for dissection.
  • Comparison microscope, which has two separate light paths allowing direct comparison of two samples via one image in each eye.
  • Inverted microscope, for studying samples from below; useful for cell cultures in liquid, or for metallography.
  • Fiber optic connector inspection microscope, designed for connector end-face inspection
  • Traveling microscope, for studying samples of high optical resolution.

Other microscope variants are designed for different illumination techniques:

  • Petrographic microscope, whose design usually includes a polarizing filter, rotating stage and gypsum plate to facilitate the study of minerals or other crystalline materials whose optical properties can vary with orientation.
  • Polarizing microscope, similar to the petrographic microscope.
  • Phase-contrast microscope, which applies the phase contrast illumination method.
  • Epifluorescence microscope, designed for analysis of samples which include fluorophores.
  • Confocal microscope, a widely used variant of epifluorescent illumination which uses a scanning laser to illuminate a sample for fluorescence.
  • Two-photon microscope, used to image fluorescence deeper in scattering media and reduce photobleaching, especially in living samples.
  • Student microscope – an often low-power microscope with simplified controls and sometimes low quality optics designed for school use or as a starter instrument for children.[4]
  • Ultramicroscope, an adapted light microscope that uses light scattering to allow viewing of tiny particles whose diameter is below or near the wavelength of visible light (around 500 nanometers); mostly obsolete since the advent of electron microscopes
  • Tip-enhanced Raman microscope, is a variant of optical microscope based on tip-enhanced Raman spectroscopy, without traditional wavelength-based resolution limits.[5][6] This microscope primarily realized on the scanning-probe microscope platforms using all optical tools.

Digital microscope

A miniature USB microscope.

A digital microscope is a microscope equipped with a digital camera allowing observation of a sample via a computer. Microscopes can also be partly or wholly computer-controlled with various levels of automation. Digital microscopy allows greater analysis of a microscope image, for example measurements of distances and areas and quantitaton of a fluorescent or histological stain.

Low-powered digital microscopes, USB microscopes, are also commercially available. These are essentially webcams with a high-powered macro lens and generally do not use transillumination. The camera attached directly to the USB port of a computer, so that the images are shown directly on the monitor. They offer modest magnifications (up to about 200×) without the need to use eyepieces, and at very low cost. High power illumination is usually provided by an LED source or sources adjacent to the camera lens.

Digital microscopy with very low light levels to avoid damage to vulnerable biological samples is available using sensitive photon-counting digital cameras. It has been demonstrated that a light source providing pairs of entangled photons may minimize the risk of damage to the most light-sensitive samples. In this application of ghost imaging to photon-sparse microscopy, the sample is illuminated with infrared photons, each of which is spatially correlated with an entangled partner in the visible band for efficient imaging by a photon-counting camera.[7]

Other Languages
العربية: مجهر ضوئي
azərbaycanca: Optik mikroskop
brezhoneg: Mikroskop
한국어: 광학 현미경
Bahasa Indonesia: Mikroskop cahaya
македонски: Оптички микроскоп
Bahasa Melayu: Mikroskop optik
日本語: 光学顕微鏡
norsk nynorsk: Lysmikroskop
ਪੰਜਾਬੀ: ਖੁਰਦਬੀਨ
Simple English: Light microscope
српски / srpski: Оптички микроскоп
srpskohrvatski / српскохрватски: Optički mikroskop
Türkçe: Optik mikroskop