An electron microscope uses a beam of electrons to illuminate a specimen and produce a magnified image.

An electron microscope (EM) has greater resolving power than a light-powered optical microscope because electrons have wavelengths about 100,000 times shorter than visible light (photons) . They can achieve better than 50 pm resolution
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andmagnifications of up to about 10,000,000x whereas ordinary, non-confocal light microscopes

are limited by diffraction to about 200 nm resolution and useful magnifications below 2000x. The electron microscope uses electrostatic and electromagnetic "lenses" to control the electron beam and focus it to form an image. These lenses are analogous to but different from the glass lenses of an optical microscope that form a magnified image by focusing light on or through the specimen. Electron microscopes are used to observe a wide range of biological and inorganic specimens including microorganisms, cells, large molecules, biopsysamples, metals, and crystals. Industrially, the electron microscope is often used for quality control and failure analysis. Types Transmission electron microscope (TEM) Main article: Transmission electron microscope The original form of electron microscope, the transmission electron microscope (TEM) uses a high voltage electron beam to create an image. The electrons are emitted by an electron gun, commonly fitted with a tungsten filament cathode as the electron source. The electron beam is accelerated by an anode typically at +100 keV (40 to 400 keV) with respect to the cathode, focused by electrostaticand electromagnetic lenses, and transmitted through the specimen that is in part transparent to electrons and in part scatters them out of the beam. When it emerges from the specimen, the electron beam carries information about the structure of the specimen that is magnified by the objective lens system of the microscope. The spatial variation in this information (the "image") may be viewed by projecting the magnified electron image onto a fluorescent viewing screen coated with a phosphor or scintillator material such as zinc sulfide. Alternatively, the image can be photographically recorded by exposing a photographic film or plate directly to the electron beam, or a high-resolution phosphor may be coupled by means of a lens optical system or a fibre optic light-guide to the sensor of a CCD (charge-coupled device) camera. The image detected by the CCD may be displayed on a monitor or computer. Resolution of the TEM is limited primarily by spherical aberration, but a new generation of aberration correctors have been able to partially overcome spherical aberration to increase resolution. Hardware correction of spherical aberration for the high-resolution transmission electron microscopy (HRTEM) has allowed the production of images with resolution below 0.5 angstrom (50 picometres) and magnifications above 50 million times. research and development.
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The ability to

determine the positions of atoms within materials has made the HRTEM an important tool for nano-technologies

An important mode of TEM utilization is electron diffraction. The advantages of electron diffraction over X-ray crystallography are that the specimen need not be a single crystal or even a polycrystalline powder, and also that the Fourier transform reconstruction of the object's magnified structure occurs physically and thus avoids the need for solving the phase problem faced by the X-ray crystallographers after obtaining their X-ray diffraction patterns of a single crystal or polycrystalline powder. The major disadvantage of the transmission electron microscope is the need for extremely thin sections of the specimens, typically about 100 nanometers. Biological specimens typically require to be chemically fixed, dehydrated and embedded in a polymer resin to stabilize them sufficiently to allow ultrathin

sectioning. Sections of biological specimens, organic polymers and similar materials may require special `staining' with heavy atom labels in order to achieve the required image contrast. Scanning electron microscope Unlike the TEM, where electrons of the high voltage beam carry the image of the specimen, the electron beam of the scanning electron microscope (SEM)
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does not at any time carry a complete image of the specimen. The SEM

produces images by probing the specimen with a focused electron beam that is scanned across a rectangular area of the specimen (raster scanning). When the electron beam interacts with the specimen, it loses energy by a variety of mechanisms. The lost energy is converted into alternative forms such as heat, emission of low-energy secondary electrons and high-energy backscattered electrons, light emission (cathodoluminescence) or X-ray emission, which provide signals carrying information about the properties of the specimen surface, such as its topography and composition. The image displayed by an SEM maps the varying intensity of any of these signals into the image in a position corresponding to the position of the beam on the specimen when the signal was generated. In the SEM image of an ant shown at right, the image was constructed from signals produced by a secondary electron detector, the normal or conventional imaging mode in most SEMs. Generally, the image resolution of an SEM is about an order of magnitude poorer than that of a TEM. However, because the SEM image relies on surface processes rather than transmission, it is able to image bulk samples up to many centimetres in size and (depending on instrument design and settings) has a great depth of field, and so can produce images that are good representations of the three-dimensional shape of the sample. Another advantage of SEM is its variety called environmental scanning electron microscope (ESEM) can produce images of sufficient quality and resolution with the samples being wet or contained in low vacuum or gas. This greatly facilitates imaging biological samples that are unstable in the high vacuum of conventional electron microscopes. Reflection electron microscope In the reflection electron microscope (REM) as in the TEM, an electron beam is incident on a surface, but instead of using the transmission (TEM) or secondary electrons (SEM), the reflected beam of elastically scattered electrons is detected. This technique is typically coupled with reflection high energy electron diffraction (RHEED) and reflection high-energy loss spectroscopy (RHELS). Another variation is spin-polarized low-energy electron microscopy (SPLEEM), which is used for looking at the microstructure ofmagnetic domains. Scanning transmission electron microscope Main article: Scanning transmission electron microscopy The STEM rasters a focused incident probe across a specimen that (as with the TEM) has been thinned to facilitate detection of electrons scattered through the specimen. The high resolution of the TEM is thus possible in STEM. The focusing action (and aberrations) occur before the electrons hit the specimen in the STEM, but afterward in the TEM. The STEMs use of SEM-like beam rastering simplifies annular dark-field imaging, and other analytical techniques, but also means that image data is acquired in serial rather than in parallel fashion. Often TEM can be equipped with the scanning option and then it can function both as TEM and STEM. Low-voltage electron microscope The low-voltage electron microscope (LVEM) is a combination of SEM, TEM and STEM in one instrument, which operates at relatively low electron accelerating voltage of 5 kV. Low voltage reduces the specimen damage by the incident electrons and increases image contrast that is especially important for biological specimens. This increase in
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contrast significantly reduces, or even eliminates the need to stain. Sectioned samples generally need to be thinner than they would be for conventional TEM (2065 nm). Resolutions of a few nm are possible in TEM, SEM and STEM modes.
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Robert Hooke FRS (28 July [O.S. 18 July] 1635 3 March 1703) was an English natural philosopher, architect and polymath. His adult life comprised three distinct periods: as a scientific inquirer lacking money; achieving great wealth and standing through his reputation for hard work and scrupulous honesty following the great fire of 1666, but eventually becoming ill and party to jealous intellectual disputes. These issues may have contributed to his relative historical obscurity. He was at one time simultaneously the curator of experiments of the Royal Society and a member of its council, Gresham Professor of Geometry and a Surveyor to the City of London after the Great Fire of London, in which capacity he appears to have performed more than half of all the surveys after the fire. He was also an important architect of his time, though few of his buildings now survive and some of those are generally misattributed, and was instrumental in devising a set of planning controls for London whose influence remains today. Allan Chapman has characterised him as "England'sLeonardo".
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Hooke studied at Wadham College during the Protectorate where he became one of a tightly knit group of ardent Royalists centred around John Wilkins. Here he was employed as an assistant to Thomas Willis and to Robert Boyle, for whom he built the vacuum pumps used in Boyle's gas law experiments. He built some of the earliest Gregorian telescopes, observed the rotations of Mars and Jupiter and, based on his observations of fossils, was an early proponent of biological evolution.
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He investigated the phenomenon of refraction, deducing the wave

theory of light, and was the first to suggest that matter expands when heated and that air is made of small particles separated by relatively large distances. He performed pioneering work in the field of surveying and map-making and was involved in the work that led to the first modern plan-form map, though his plan for London on a grid system was rejected in favour of rebuilding along the existing routes. He also came near to deducing that gravity follows an inverse square law, and that such a relation governs the motions of the planets, an idea which was subsequently developed by Newton.
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Much of Hooke's scientific work was conducted in his capacity as curator of experiments of

the Royal Society, a post he held from 1662, or as part of the household of Robert Boyle. Antonie Philips van Leeuwenhoek (in Dutch also Anthonie, Antoni, or Theunis, inEnglish, Antony or Anton; levnhk/, Dutch: [lenhuk] ( listen); October 24, 1632 August 26, 1723) was a Dutch tradesman
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and scientist from Delft, Netherlands. He is commonly known as "the Father of Microbiology", and considered to be the firstmicrobiologist. He is best known for his work on the improvement of the microscopeand for his contributions towards the establishment of microbiology. Using his handcrafted microscopes, he was the first to observe and describe single-celled organisms, which he originally referred to as animalcules, and which we now refer to asmicroorganisms. He was also the first to record microscopic observations of musclefibers, bacteria, spermatozoa, and blood flow in capillaries (small blood vessels). Van Leeuwenhoek did not author any books, although he did write many letters. Robert Brown FRSE FRS FLS MWS (born 21 December 1773 and died 10 June 1858) was a Scottish botanist and palaeobotanist who made important contributions to botany largely through his pioneering use of the microscope. His contributions include one of the earliest detailed descriptions of the cell nucleus and cytoplasmic streaming; the first observation ofBrownian motion; early work on plant pollination and fertilisation, including being the first to recognise the fundamental difference between gymnosperms and angiosperms; and some of the earliest studies

in palynology. He also made numerous contributions to plant taxonomy, including the erection of a number of plant families that are still accepted today; and numerous Australian plant genera and species, the fruit of his exploration of that continent with Matthew Flinders. What is the largest cell? Largeness refers to size, not weight, so the ostrich egg is definitely not the largest biological cell. The first type of cell larger than the ostrich egg are nerve cells in especially long animals, such as the Giant Squid and Colossal Squid, which may have nerve cells as long as 12 m (39 ft). This is about 80 times larger than an ostrich egg. Nerve cells have very long axons (protrusions), enabling the brain to send signals to distant limbs almost instantly. In giraffes, the nerve cells may be several meters long, running the whole length of the animal's neck, and in humans, the longest nerve cells are about 1.5 m, running from the base of the spine to the toes. So even the human body has biological cells larger than the ostrich egg. In terms of volume, the largest and smallest cells in the human body are the gametes, or the sex cells. The female gamete, the oocyte (also known as the egg or the ovum), is about 1000 micrometers, or one millimeter, in diameter, which is just visible to the naked eye without the aid of a microscope or other magnification device. It needs this size as nourishment before it implants itself in the uterus. The male gamete, the spermatozoan (also known as the sperm cell), is only about 60 micrometers long, and therefore is the smallest cell of the human body.

Why are cells small? All the answers Ive seen so far are missing the point. The answer to your question has to do with the surface-tovolume ratio. As a cell gets bigger (no matter what the shape of the cell is, being spherical, cuboidal, columnar, what have you...) the volume of the cell gets bigger FASTER than the surface area of the cell. The surface area is the outermost part of the cell and in the real world, this represents the cell membrane. This is bad news for a cell. Why? First you have to realize that the cell membrane is responsible for transporting things into and out of the cell. Basically then, the inside volume of the cell DEPENDS UPON the cell membrane to provide it with necessary nutrients so it can function. If the volume grows faster than the cell membrane (as the cell gets bigger), the the cell membrane's ability to function as a provider for the interior of the cell is greatly compromised. The cell solves this by dividing, to keep its overall cell size relatively small. As far as the agar experiment goes, I dont know but I can give you an idea. Basically, it sounds like a PROOF of the logic I have given you above. I imagine that if you cut cubes of agar into different sizes (say a quarter inch cube, half inch, inch and so on) and if you dissolve those cubes in a solvent you will find that they dissolve at different rates. The length of time that is required to dissolve the cube will be proportional to the VOLUME of the cube. What you should find is this: even though the quarter inch cube has a side that is exactly half of the half inch cube, it will take MORE THAN DOUBLE the amount of time for the half inch cube to dissolve (compared to the quarter inch). Recall that these cubes represent a cell that has grown. The reason for the prolonged dissolution time is this: as a cell grows, the surface area increases as the square (area is given in square units), whereas the volume increases as the cube (volume is given in cubic units). Since dissolution time is reflective of the volume of the cell, your expectation should be that if you double the area, it will take MORE THAN DOUBLE the amount of time to dissolve the cube since the volume is more than doubled. Try it and see! Again, this is why cells must remain small. So that the cell membrane can adequately provide the interior of the cell with nutrients.

Cell theory refers to the idea as cells are the basic unit of structure in every living thing. Development of this theory during the mid 17th century was made possible by advances in microscopy. This theory is one of

the foundations ofbiology. The theory says that new cells are formed from other existing cells, and that the cell is a fundamental unit of structure, function and organization in all living organisms. History The cell was discovered by Robert Hooke in 1665. He examined (under a coarse, compound microscope) very thin slices of cork and saw a multitude of tiny pores that he remarked looked like the walled compartments a monk would live in. Because of this association, Hooke called them cells, the name they still bear. However, Hooke did not know their real structure or function. Hooke's description of these cells (which were actually non-living cell walls) was published in Micrographia. most living cells. The first person to make a compound microscope was Zacharias Jansen, while the first to witness a live cell under a microscope was Antonie van Leeuwenhoek, who in 1674 described the algae Spirogyra and named the moving organisms animalcules, meaning "little animals".
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His cell observations gave no indication of thenucleus and other organelles found in

Leeuwenhoek probably also saw bacteria.

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Cell theory was in

contrast to the vitalismtheories proposed before the discovery of cells. The idea that cells were separable into individual units was proposed byLudolph Christian Treviranus Jacob Paul Moldenhawer.
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and Johann

All of this finally led to Henri Dutrochet formulating one of the fundamental tenets of
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modern cell theory by declaring that "The cell is the fundamental element of organization".

The observations of Hooke, Leeuwenhoek, Schleiden, Schwann, Virchow, and others led to the development of the cell theory. The cell theory is a widely accepted explanation of the relationship between cells and living things. The cell theory states:

All living things or organisms are made of cells and their products. New cells are created by old cells dividing into two. Cells are the basic building units of life.

The cell theory holds true for all living things, no matter how big or small, or how simple or complex. Since according to research, cells are common to all living things, they can provide information about all life. And because all cells come from other cells, scientists can study cells to learn about growth, reproduction, and all other functions that living things perform. By learning about cells and how they function, you can learn about all types of living things. Credit for developing cell theory is usually given to three scientists: Theodor Schwann, Matthias Jakob Schleiden, and Rudolf Virchow. In 1839, Schwann and Schleiden suggested that cells were the basic unit of life. Their theory accepted the first two tenets of modern cell theory (see next section, below). However, the cell theory of Schleiden differed from modern cell theory in that it proposed a method of spontaneous crystallization that he called "free cell formation".
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In 1855, Rudolf Virchow concluded that all cells come from pre-existing cells, thus completing the
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classical cell theory. (Note that the idea that all cells come from pre-existing cells had in fact already been proposed by Robert RemakVirchow is considered to have plagarised Remak. ) Modern interpretation The generally accepted parts of modern cell theory include: 1. All known living things are made up of one or more cells. 2. All living cells arise from pre-existing cells by division.

3. The cell is the fundamental unit of structure and function in all living organisms. 4. The activity of an organism depends on the total activity of independent cells. 5. Energy flow (metabolism and biochemistry) occurs within cells. 6. Cells contain hereditary information (DNA) which is passed from cell to cell during cell division. 7. All cells are basically the same in chemical composition in organisms of similar species.

Cell fractionation Cell fractionation is the separation of homogeneous sets, usually organelles, from a population of cells. Steps There are three principal steps involved: 1. Disruption (homogenization) of cells and liberation of organelles. 2. Macro Filtration 3. Purification of cell components. [edit]Homogenization Tissue is typically homogenized in an isotonic buffer solution using a variety of mechanisms. A 'Potter-Elvehjem homogeniser' is often used as it is relatively gentle. Other procedures include grinding, mincing, chopping, pressure changes, osmotic shock, freeze-thawing, and ultra-sound homogenization. The solution is homogenized in an isotonic solution to stop osmotic damage, with a pH buffer to regulate pH, and at an ice-cold temperature to prevent enzyme damage. The organelles are kept either cold, isotonic or buffered. See Cell disruption for further details. [edit]Filtration This step may not be necessary depending on the source of the cells. Animal tissue however is likely to yield connective tissue which must be removed. Commonly, filtration is achieved either by pouring through gauze or with a suction filter and the relevant grade ceramic filter. [edit]Purification Invariably achieved by Differential centrifugation - the sequential increase in gravitational force resulting in the sequential separation of organelles according to their density.

Cell disruption is a method or process for releasing biological molecules from inside a cell. Differential centrifugation is a common procedure in microbiology and cytology used to separate certain organelles from whole cellsfor further analysis of specific parts of cells. In the process, a tissue sample is first homogenised to break the cell membranes and mix up the cell contents. The homogenate is then subjected to repeated centrifugations, each time removing the pellet and increasing thecentrifugal force. Finally, purification may be done through equilibrium sedimentation, and the desired layer is extracted for further analysis. Separation is based on size and density, with larger and denser particles pelleting at lower centrifugal forces. As an example, unbroken whole cells will pellet at low speeds and short intervals such as 1,000g for 5 minutes. Smaller cell

fragments and organelles remain in the supernatant and require more force and greater times to pellet. In general, one can enrich for the following cell components, in the separating order in actual application:

Whole cells and nuclei; Mitochondria, lysosomes and peroxisomes; Microsomes (vesicles of disrupted endoplasmic reticulum); and Ribosomes and cytosol.