Optical microscope

The optical microscope, often referred to as the light microscope, is a type of microscope that
uses visible light and a system of lenses to magnify images of small subjects. 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 image from an optical microscope can 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.
Alternatives to optical microscopy which do not use visible light include scanning electron
microscopy and transmission electron microscopy and Scanning probe microscopy.
On 8 October 2014, the Nobel Prize in Chemistry was awarded to Eric Betzig, William
Moerner and Stefan Hell for "the development of super-resolved fluorescence microscopy," which
brings "optical microscopy into the nanodimension"

Types

Diagram of a simple microscope

There are two basic types of optical microscopes: simple microscopes and compound microscopes.
A simple microscope is one which uses a single lens for magnification, such as a magnifying glass. A
compound microscope uses several lenses to enhance the 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.[3][4] 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).[5]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.[5] 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:

 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. [6]
 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
Digital microscope

A miniature USB microscope.

Main article: Digital 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]
 Components

Basic optical transmission microscope elements (1990s)

All modern optical microscopes designed for viewing samples by transmitted light share the same
basic components of the light path. In addition, the vast majority of microscopes have the same
'structural' components[28] (numbered below according to the image on the right):

 Eyepiece (ocular lens) (1)

 Objective turret, revolver, or revolving nose piece (to hold multiple objective lenses) (2)
 Objective lenses (3)
 Focus knobs (to move the stage)
 Coarse adjustment (4)
 Fine adjustment (5)
 Stage (to hold the specimen) (6)
 Light source (a light or a mirror) (7)
 Diaphragm and condenser (8)
 Mechanical stage (9)
Eyepiece (ocular lens)[edit]
Main article: Eyepiece
The eyepiece, or ocular lens, is a cylinder containing two or more lenses; its function is to bring the
image into focus for the eye. The eyepiece is inserted into the top end of the body tube. Eyepieces
are interchangeable and many different eyepieces can be inserted with different degrees of
magnification. Typical magnification values for eyepieces include 5×, 10× (the most common), 15×
and 20×. In some high performance microscopes, the optical configuration of the objective lens and
eyepiece are matched to give the best possible optical performance. This occurs most commonly
with apochromatic objectives.
Objective turret (revolver or revolving nose piece)
Objective turret, revolver, or revolving nose piece is the part that holds the set of objective lenses. It
allows the user to switch between objective lenses.

Operation

Optical path in a typical microscope

The optical components of a modern microscope are very complex and for a microscope
to work well, the whole optical path has to be very accurately set up and controlled. Despite this, the
basic operating principles of a microscope are quite simple.
The objective lens is, at its simplest, a very high-powered magnifying glass, i.e. a lens with
a very short focal length. This is brought very close to the specimen being examined so that the light
from the specimen comes to a focus about 160 mm inside the microscope tube. This creates an
enlarged image of the subject. This image is inverted and can be seen by removing the eyepiece and
placing a piece of tracing paper over the end of the tube. By carefully focusing a brightly lit specimen,
a highly enlarged image can be seen. It is this real image that is viewed by the eyepiece lens that
provides further enlargement.
In most microscopes, the eyepiece is a compound lens, with one component lens near the
front and one near the back of the eyepiece tube. This forms an air-separated couplet. In many
designs, the virtual image comes to a focus between the two lenses of the eyepiece, the first lens
bringing the real image to a focus and the second lens enabling the eye to focus on the virtual
image.[30]