SEMINAR ON SCANNING ELECTRON MICROSCOPE

SUJIT SEKHAR MOHARANA

This seminar is intended to give a fair idea about the various principles, working and the instrumentation of scanning electron microscope.

INDIAN INSTITUTE

OF

TECHNOLOGY

KHARAGPUR

9/14/2012

CONTENTS
1. HISTORY 2. INTRODUCTION 3. PHYSICS BEHIND 4. SEM DEMYSTIFIED 5. SAMPLE PREPARATION 6. EFFECT OF SOME PARAMETERS 7. APPLICATIONS

HISTORY
The earliest known work describing the concept of a Scanning Electron Microscope was by M. Knoll (1935) who, along with other pioneers in the field of electron optics, was working in Germany. The first SEM used to examine the surface of a solid specimen was described by Zworykin et al. (1942), working in the RCA Laboratories in the United States. The electron optics of the instrument consisted of three electrostatic lenses with scan coils placed between the second and third lenses; the electron gun was located at the bottom so the specimen chamber was at a comfortable height for the operator. This was a common practice in the early days. It did suffer from the slight problem however, that the specimen might fall down the column. A resolution of about 50 nm was achieved with this first SEM. By comparison with the rapidly developing TEM, this figure was considered unexciting and further development lapsed. In the late 1940s C. W. Oatley, then a lecturer in the Engineering Department of Cambridge University, England, became interested in conducting research in the field of electron optics and decided to re-investigate the SEM as a complement to the work being done on the TEM by V. E. Cosslett, also in Cambridge at the Physics Department. One of Oatley's students, Ken Sander, began work on a column for a transmission electron microscope using electrostatic lenses, but Ken took ill after about one year and had to leave for a time. This work then was taken up by Dennis McMullan in 1948, and he and Oatley built their first SEM. By 1952 this instrument had achieved a resolution of 50 nm, but by far the most important thing about this SEM is that it produced the first micrographs showing the striking three-dimensional imaging characteristics of the modern-day SEM (McMullan (1953)). Dennis McMullan was followed by Ken Smith (started 1952) who took over SEM1, made a number of improvements to the electron optical system, and improved the efficiency of secondary electron collection. He showed for the first time that a stable image could be formed using the true low-energy component of the total secondary emission. Ken has told me personally that during this period in the development of the SEM, microscopists in general showed virtually no interest in the instrument, and he and Oatley were continually looking for applications to promote the SEM; many which they tried were never published. However, they published the first paper setting out clearly the fields of application for the SEM (Smith and Oatley (1955)).

The third research student on the SEM under Oatley's supervision was O. C. Wells (started 1953), who built a second SEM, also incorporating electrostatic lenses; unlike SEM1, however, this instrument had the gun at the bottom of the column - a configuration considered better for experimental work. All SEMs built in the Engineering Department subsequently conformed to this layout. Wells pioneered the use of the scintillator backscattered (BSE) detector (as an alternative to the secondary electron multiplier used in SEM1) and applied his SEM to many new types of specimen including a large-scale study of fibers. He was also the first to use stereographic pairs to produce SEM micrographs with quantifiable depth information (Wells (1960)). The next important step was taken by Oatley's fourth research student, Everhart (started 1955), who improved the secondary electron (SE) detector by using a scintillator to convert electrons to photons, which were then transmitted by a light pipe directly interfaced to the photomultiplier tube. This idea was followed up by Thornley (started 1957), and their ground-breaking work resulted in the publication of a much-quoted paper:- "Wide-band detector for micromicroampere low-energy electron currents" (Everhart and Thornley (1960)). Replacement of the electron multiplier with the new scintillator/photomultiplier combination increased the amount of signal collected and resulted in an improvement in signal-to-noise ratio. Hence, weak contrast mechanisms, such as voltage contrast (discovered by Oatley and Everhart (1957)) could be better investigated. The term voltage contrast" evolved from the fact that as the voltage applied to a specimen was changed, the image contrast changed. (Tom Everhart and Oatley were investigating this one day and Tom said: Well, voltage contrast!" and the term have stuck to this day). Image interpretation was also improved in this early period of research when both Everhart and Wells made the first quantitative studies of the effects of beam penetration on image formation in the SEM. It was always one of Oatley's great ambitions to produce and market a really simple low-cost SEM; he argued that most of the work undertaken by microscopists did not require high resolution. To further this idea Peter Spreadbury, his fifth student (started 1956), built a simple SEM utilizing a CRT as a display unit. New fields of application were opened up by Gary Stewart (started 1958) who fitted an ion gun to the SEM specimen chamber to allow ion bombardment of the specimen. After his research, Gary went on to the Cambridge Instrument

Company to pioneer the production of the Stereoscan. The ion beam work was later extended by Alec Broers (started 1961) who improved the ion beam optics of the instrument and added a magnetic objective lens to improve resolution. He used this set-up to conduct some of the earliest experiments in electron beam micro-fabrication. Another breakthrough was achieved by Haroon Ahmed (started 1959) who modified the SEM built by Wells (SEM2) to enable the examination of thermionic emitters at temperatures exceeding 1000K. The first SEM in the Department to achieve a resolution of 10 nm was built by Fabian Pease (started 1960). This was an all magnetic lens SEM, the fifth to be constructed in the Group. Several of these SEMs were later manufactured in the Departmental Workshops and used by other Groups within the University. A brief list of all the works taken up by students of Prof Oatleys group towards the development of electron microscopes is presented here. 1. D McMullan: 1948. Constructed SEM1 and produced the first micrographs showing the three-dimensional topographic image formation characteristic of modern-day instruments. Theoretical analysis of probe formation; measurements of distribution of backscattered electrons with angle; considered mechanisms of BSE contrast formation. Dissertation: "Investigations relating to the Design of Electron Microscopes" 1952. 2. K C A Smith: 1952. Made improvements to SEM1, including efficient detection of the low-energy secondary electron (SE) component; considered SE contrast formation; extended theory of probe formation; investigated a wide range of applications; dynamic experiments, including chemical reactions at elevated temperatures. Dissertation: "The Scanning Electron Microscope and its Fields of Application" 1956. 3. O C Wells: 1953. Constructed SEM2 and applied it to study of fibers. Explored new types of detector; used scintillator/photomultiplier detector in many novel configurations; established theory of stereomicroscopy in the SEM; investigated ways of examining non-conducting specimens, including the use of positive ion bombardment; investigated atomic number contrast. Dissertation: "The construction of a Scanning Electron Microscope and its application to the study of fibers" 1957.

4. T E Everhart: 1955. Continued improvements to SEM1; devised new detector (Everhart-Thornley detector). Studied contrast mechanisms in detail, including potential contrast; developed a new theory of reflection of electrons from solids. Dissertation: Microscope" 1958. "Contrast formation in the Scanning Electron

5. P J Spreadbury: 1956. Constructed a simple SEM using a cathode-ray oscillograph as the display unit; made careful measurements of the performance of the electron gun. Constructed numerous pieces of electronic equipment which were used in other SEMs and projects in the laboratory. Dissertation: "Investigations relating to the design of a Simple Scanning Electron Microscope" 1958. 6. R F M Thornley 1957. Made improvements to SEM2. Conducted detailed study of Everhart detector. Low-voltage operation for examination of non-conducting specimens; studies of frozen biological specimens. Dissertation: "New applications of the Scanning Electron Microscope" 1960. 7. A D G Stewart 1958. Completed construction of a new microscope begun by Oatley (SEM4). Addition of ion gun allowed the direct observation of specimens while undergoing sputtering. Later moved to Cambridge Instrument Co. to work on Stereoscan project (see section 5). Dissertation: not submitted. 8. H Ahmed 1959 (supervisor A H W Beck). Used SEM2 to investigate activation processes of dispenser cathodes. Direct observation of emitting cathodes at temperatures exceeding 1300 K. Dissertation: "Studies on high-current-density Thermionic Cathodes" 1962. 9. R F W Pease 1960. Designed a new microscope (SEM5) - first SEM to achieve a resolution of 10 nm. Several of these instruments were made in the Engineering Dept. and used in other Groups. Also supplied to F P Bowdens Group in the Cavendish Laboratory (see section 5).

Dissertation: "High resolution Scanning Electron Microscope" 1963. 10. An N Broers 1961. Made improvements to SEM4, including addition of magnetic lens. Added mass-filter to ion beam system to obtain pure ion beam species. Found that surface contaminants affected rates of sputtering and could thus be used to mask selected areas of the specimen surface. Used this effect to lay down patterns of gold wires and other structures - one of the earliest successful attempts at electron beam microfabrication and micromachining. Dissertation: "Selective ion beam etching in the Scanning Electron Microscope" 1965.

COMMERCIAL DEVELOPMENT Up to the mid-1950s there was little interest in the work of Oatleys Group, but a turning point in attitudes towards scanning electron microscopy came when D Atack, a member of the Pulp and Paper Research Institute of Canada (PPRIC), and J H L McAuslan, with Imperial Chemical Industries, learned of the work on the SEM. Both were on sabbatical leave at the time in F P Bowden's Group in the Cavendish, and they decided to explore the potential of the instrument for their work. Using SEM1 Atack examined a range of pulp and paper specimens, while McAuslan studied the thermal decomposition of silver azide crystals. This work generated strong interest at the PPRIC, and it was subsequently arranged that the Institute would finance the construction of a new SEM in the Engineering Department. Smith undertook the task of designing this new instrument - designated SEM3 - as a post-doctoral research project in 1956. At this stage Oatley persuaded Associated Electrical Industries (AEI) (formerly Metropolitan Vickers), a company then manufacturing both transmission instruments and electron probe micro-analyzers, to take an interest in the SEM. An understanding was reached that if the microscope appeared to be commercially viable, AEI would take it up. Later, in 1958, AEI received an order from Bowden, but for commercial reasons the instrument produced was based on their current production micro-analyzers, little use was made of the experience gained from the construction and operation of SEM3, and it was not successful. Bowden returned the microscope to AEI and later ordered a copy of the Pease-Nixon microscope from the Engineering Department. This ended AEI's attempt to enter the SEM market (Agar 1996; Brown et. al. 1996).

SEM3 was shipped to Canada in 1958 and used successfully by the PPRIC in their Montreal laboratories and by other companies which hired time on the instrument, including the du Pont chemical company of the USA. This, together with an accelerated flow of results from members of Oatley's Group - Nixon, Thornley, Stewart, Ahmed, Pease, and Broers - produced a change in attitudes towards the SEM as its advantages became better appreciated. In 1961 Nixon and Smith made an informal approach to the Cambridge Instrument Company, suggesting that the Company should take up manufacture of the SEM as well as the micro-analyzers, which they were marketing at the time. Shortly afterwards Oatley reached a formal agreement with H C Pritchard, the managing director of the Company, and arrangements were made for the manufacture of two prototypes, one of which went to the du Pont Company (Fig. 4). In 1962 one of Oatley's former research students, A D G Stewart, joined the Company to take over development of the new SEM, and with Government backing, a batch of five microscopes was manufactured in 1965.

INTRODUCTION
Electron Microscopes are scientific instruments that use a beam of highly energetic electrons to examine objects on a very fine scale. This examination can yield information about the topography (surface features of an object), morphology (shape and size of the particles making up the object), composition (the elements and compounds that the object is composed of and the relative amounts of them) and crystallographic information (how the atoms are arranged in the object). Electron Microscopes were developed due to the limitations of Light Microscopes which are limited by the physics of light to 500x or 1000x magnification and a resolution of 0.2 micrometers. In the early 1930's this theoretical limit had been reached and there was a scientific desire to see the fine details of the interior structures of organic cells (nucleus, mitochondria...etc.). This required 10,000x plus magnification which was just not possible using Light Microscopes. The Transmission Electron Micros cope (TEM) was the first type of Electron Microscope to be developed and is patterned exactly on the Light Transmission Microscope except that a focused beam of electrons is used instead of light to "see through" the specimen. It was developed by Max Knoll and Ernst Ruska in Germany in 1931. The first Scanning Electron Microscope (SEM) debuted in 1942 with the first commercial instruments around 1965. Its late development was due to the electronics involved in "scanning" the beam of electrons across the sample. Electron Microscopes (EMs) function exactly as their optical counterparts except that they use a focused beam of electrons instead of light to "image" the specimen and gain information as to its structure and composition. The basic steps involved in all EMs are the following:A stream of electrons is formed in high vacuum (by electron guns). This stream is accelerated towards the specimen (with a positive electrical potential) while is confined and focused using metal apertures and magnetic lenses into a thin, focused, monochromatic beam. The sample is irradiated by the beam and interactions occur inside the irradiated sample, affecting the electron beam. These interactions and effects are detected and transformed into an image. There are three basic ways in which an image can be formed. These are:1. The simplest to imagine is the projection mode in which an image is formed when an object is kept in front of an illuminated source. 2. The second type is the optical mode which is formed by the conventional lens systems. One particular thing to note is that in both these modes images are formed in parallel i.e. all parts of images are formed almost simultaneously.

3. The third mode of image formation is the scanning mode in which each point of the picture is presented serially. The best known example is the image formed by a television picture tube which forms the image at such a high frequency that our eyes perceive them as a whole. In this seminar we will discuss an instrument which has revolutionized the field of science and has attached a new dimension to the meaning of observation and measurement using this mode of image formation. This instrument is the SCANNING ELECTRON MICROSCOPE.

PHYSICS BEHIND
When an electron beam interacts with the atoms in a sample, individual incident electrons undergo two types of scattering 1. elastic scattering 2. inelastic scattering In the elastic scattering, only the trajectory changes and the kinetic energy and velocity of the incident electrons remain constant. In the case of inelastic scattering, some incident electrons will actually collide with and displace electrons from their orbits (shells) around nuclei of atoms comprising the sample. These interaction places the atom in an excited (unstable) state. Specimen interaction is what makes Electron Microscopy possible. The interactions (inelastic) noted on the diagram are utilized when examining thick or bulk specimens (Scanning Electron Microscopy,SEM).

REACTIONS EXPLOITED IN SEM

1.

SECONDARY ELECTRONS:- When a sample is bombarded with

electrons, the strongest region of the electron energy spectrum is due to secondary electrons. The secondary electron yield depends on many factors, and is generally higher for high atomic number targets, and at higher angles of incidence. Secondary electrons are produced when an incident electron excites an electron in the sample and loses most of its energy in the process. The excited electron moves towards the surface of the sample undergoing elastic and inelastic collisions until it reaches the surface, where it can escape if it still has sufficient energy. Production of secondary electrons is very topography related. Due to their low energy (5eV) only secondary electrons that are very near the surface (<10 nm) can exit the sample and be examined. Any changes in topography in the sample that are larger than this sampling depth will change the yield of secondary electrons due to collection efficiencies. Collection of these electrons is aided by using a "collector" in conjunction with the secondary electron detector.

2.

BACKSCATTERED

Backscattered electrons consist of high-energy electrons originating in the electron beam that are reflected or back-scattered out of the specimen interaction volume. The production of backscattered electrons varies directly with the specimen's atomic number. This differing production rates causes higher atomic number elements to appear brighter than lower atomic number elements. This interaction is utilized to differentiate parts of the specimen that have different average atomic number.

ELECTRONS:-

3.

RELAXATION OF EXCITED ATOMS:- Inelastic scattering,

places the atom in an excited (unstable) state. The atom wants to return to a ground or unexcited state. Therefore, at a later time the atoms will relax giving off the excess energy. Ways of relaxation are:1. X-rays 2. Cathodoluminescence 3. Auger electrons The relaxation energy is the fingerprint of each element. When the sample is bombarded by the electron beam of the SEM, electrons are ejected from the atoms on the specimens surface. A resulting electron vacancy is filled by an electron from a higher shell, and an X-ray is emitted to balance the energy difference between the two electrons.

The EDS X-ray detector (also called EDS or EDX) measures the number of emitted x-rays versus their energy. The energy of the x-ray is characteristic of the element from which the x-ray was emitted. In practice, EDS (or EDX) is most often used for qualitative elemental analysis. Cathodoluminescence (CL) is the emission of photons of characteristic wavelengths from a material that is under high-energy electron bombardment. The electron beam is typically produced in an electron microprobe (EPMA) or scanning electron microscope (SEMCL). Auger electrons are electrons ejected by radiation-less excitation of a target atom by the incident electron beam. When an electron from the L shell drops to fill a vacancy formed by K-shell ionization, the resulting X-ray photon with energy EK - EL may not be emitted from the atom. If this photon strikes a lower energy electron (e.g.an M-shell electron), this outer electron may be ejected as a lower energy Auger electron. Auger electrons are characteristic of the fine structure of the atom and have energies between 280 eV (carbon) and 2.1keV (sulfur). By discriminating between Auger electrons of various energies, a chemical analysis of the specimen surface can be made. Auger electrons are exploited in Auger Electron Spectroscopy tools (AES). The volume inside the specimen in which interactions occur while being struck with an electron beam is called specimen interaction volume.

INTERACTION VOLUME

The combined effect of the elastic and inelastic interactions is to limit the penetration of the beam into the solid. The region of interaction between the solid and the beam is known as the interaction volume. The interaction volume is influenced by: (a) the beam energy (b) the atomic number of the solid (c) surface tilt (d) density of the solid. Now we will discuss each of these factors in detail. 1. Volume of excitation:- The size and shape of the interaction volume is limited by two factors: (a) energy loss through inelastic interactions (b) electron loss or backscattering through elastic interactions.

The resulting excitation volume is a hemispherical to jug-shaped region with the neck of jug at the specimen surface. The depth of electron penetration of an electron beam and the volume of sample with which it interacts are a function of its angle of incidence, the magnitude of its current, the accelerating voltage, and the average atomic number (Z) of the sample. Electron penetration generally ranges from 1-5 m with the beam incident perpendicular to the sample. 2. Influence of atomic number:(a) In specimens of high atomic number the electrons undergo more elastic scattering per unit distance and the average scattering angle is greater. (b) The electron trajectory in the high atomic number materials tends to deviate out of the initial direction of travel more quickly. (c) The shape of the interaction volume is greatly affected by the atomic number of the specimen. 3. Influence of tilt: - As the angle of tilt of the specimen surface increases, the interaction volume becomes smaller and asymmetric.

SEM DEMYSTIFIED
In this section we will have a brief description of the various constituents of SEM and the basic physics and working principles behind them. The basic working constituents of SEM are:1. 2. 3. 4. 5. Electron sources Magnetic lenses Scan coils Aperture lens Electron detector

ELECTRON SOURCES

Basically there are two types of emitters used in scanning electron microscope. These are the:1. Thermionic emission type 2. Field emission guns THERMIONIC EMISSION TYPE: - There are basically two variants of these used currently- tungsten filament and the lanthanum hexaboride. a thermionic emitter consists of three major components; an electron source to replenish emitted electrons, a heating element, and the emitter material itself. These thermionic emitters operate on principal that as certain materials are heated the electrons in the outer orbitals become unstable and are more likely to fly free of their atoms. These lost electrons are replaced by an electron source that is also attached to the emitter. The ability to give up electrons is related to a material's "work function." A LaB6 gun consists of a finely pointed crystal of lanthanum hexaboride. When heated by surrounding ceramic heaters electrons are emitted from the tip of this crystal. The lost electrons are replaced by an electron source. Because the size and geometry of this region of electron source is smaller than with a standard tungsten filament electron produced from a LaB6 gun are more likely to actually make it all the way to the sample

FIELD EMISSION GUNS:- A field emission gun operates on a different principle than does a thermionic emitter. Like a LaB6 or pointed tungsten filament the field emission gun uses a finely tipped tungsten crystal. The difference however is that the electron source is not heated to remove electrons and for this reason is often referred to as being "cold" sources. Instead electrons are drawn from the filament tip by an intense potential field set up by an anode that lies beneath the tip of the filament. Electrons are then pulled from a very small area of the pointed tip and proceed down the column. Often this is aided by a second anode that lies beneath the first. Acting like an electrostatic lens the two anodes serve to further coalesce and demagnify the beam. The lost electrons are replenished by an electron source attached to the tungsten tip. A primary electron beam generated by a field emission source offers significant advantages over those produced by by thermionic emitters. Because of the smaller initial spot size [< 2.0 nm Vs. 4.0-8.0 nm], and lower accelerating voltage [2-5 KV vs. 15-20 KV] a much smaller primary excitation zone is produced. Ultimately this results in much greater resolution than is capable with a conventional SEM using a tungsten filament or LaB6 crystal.

MAGNETIC LENSES
The lens system consists of:1. Condenser lens 2. objective lens 3. scan coils

WORKING OF MAGNETIC LENSES An electromagnetic lens usually consists of a solenoid (coil), surrounded by a soft iron casing (shroud) to enhance the field and can also include soft iron pole pieces that further concentrate the field. Electrons travelling at angles to the field lines experience magnetic forces, which cause them to spiral around the field lines as they move through the magnetic lens. The radius of the spiral depends on the field strength. Electrons that enter the lens along the central axis of the solenoid will initially diverge due to repulsion from other electrons and then be refocused further along the lens. The current supplied to the solenoid controls the effective shape of the lens and therefore the focal length of the lens. The focal point, called the cross-over point, is where the electrons converge to a sharp focus. The diagram below shows an electromagnetic lens.

Both electromagnetic and permanent magnetic lenses can be designed to produce various different shaped magnetic fields as required. The diagram below shows a triangular magnetic field of uniform intensity. The triangular shape means that electrons that are furthest from the axis are in the magnetic field for more time and are consequently refracted at a greater angle than

electrons near the axis. This results in the electrons in the beam being focused to a point on the axis beyond the magnetic lens

If a rectangular shaped field is used, the electron beam can be focused by making the field non-uniform as shown below. Since the magnetic field strength increases with distance from the axis, the electrons near the axis are refracted through small angles, while those further away from the axis experience greater angles of refraction.

CONDENSOR LENS It is an electromagnetic lens located just below the electron gun to focus the electron beam emitted by the filament. The condenser lens controls the intensity of the electron beam reaching the specimen. The objective lens brings the

electron beam into focus (de-magnifies) on the specimen. The electron beam is divergent after passing through the anode plate and must be collimated by condenser lenses and apertures into a relatively parallel stream. The condenser lens controls the amount of current that passes down the rest of the column by focusing the electron beam to variable degrees. The sharper the focus, the less of the beam intercepted by an aperture (small hole) located below the lenses and the higher the current. OBJECTIVE LENS The objective or "probe-forming" lens is located at the base of the column just above the sample. This lens focuses the electron beam onto the sample and controls its final size and position. The Cameca MBX microprobe uses a "minicoil" objective lens, which allows the light optical system and scanning coils to fit inside it. The magnet is asymmetrical with less of the magnet located below the air gap to limit the amount of magnetic field at the sample. REFER IMAGE BELOW (The light optics (4) and scanning coils (1) are located inside the minicoil probe-forming lens (2) at the base of the electron column. The pole piece (7) is one solid piece of metal and protects the sample from stray magnetic fields. The x-ray beams (3) are collimated by small apertures (6), and pass through an electron trap (5) that prevents backscattered electrons from entering the x-ray spectrometers.)

SCANNING COILS Deflection of the electron beam is used to scan the beam (raster) across the surface of the sample for imaging. This is accomplished using deflection coils located within the objective lens. The deflection coils consist of four radially oriented magnets arranged so that the fields are perpendicular to the optical axis. The electron beam may be scanned systematically over the sample surface for

imaging by varying the current through these lenses. Deflecting the beam off axis introduces additional aberrations that cause the beam diameter to deteriorate, and deviations from linearity in X and Y increase as the amount of deflection increases. These effects limit the maximum field or deflection size that can be used.

ELECTRON DETECTOR A scintillator-photomultiplier detector or "Everhart- Thornley" detector is used to detect the secondary electrons emitted from the specimen. The low energy secondary electrons are emitted from the sample in all directions and possess relatively low energies. They are initially gathered by a charged collector grid (or cage), which can be biased from -50 to +300 V. This draws the secondary electrons towards the scintillator, which is a thin plastic disk coated with a short-persistence phosphor that is highly efficient at converting the energy contained in the electrons into ultraviolet light photons (4000 ). The response time of the phosphor is fast and permits high resolution scanning. The outer layer of the scintillator is coated with a thin layer [10-50 nm] of aluminum, positively biased at approximately 10 KeV, which accelerates the electrons to the scintillator surface. The charged collector grid, in addition to collecting secondary electrons from the sample, helps to alleviate some of the negative effects of the scintillator aluminum layer bias which can actually distort the incident beam.

The topographical aspects of a secondary electron image depend on how many of electrons actually reach the detector. When the incident electron beam intersects the edges of topographically high portions of a sample at lower angles, it puts more energy into the volume of secondary electron production. Thus, high points produce more secondary electrons, generating a larger signal. Faces oriented towards the detector also generate more secondary electrons. Secondary electrons that are prevented from reaching the detector do not contribute to the final image and these areas will appear as shadows or darker in contrast than those regions that have a clear electron path to the detector. It should be noted that those backscattered electrons directed at the scintillator will also contribute to the signal that reaches the scintillator and form part of the the secondary electron image.

SAMPLE PREPARATION
Generally samples can be divided into two categories. These are:1. Metal or material samples 2. Biological samples Below we will have a brief description of both of these kind of samples for examination under an scanning electron microscope. METAL OR MATERIAL SAMPLES:- For conventional imaging in the SEM, specimens must be electrically conductive, at least at the surface, and electrically grounded to prevent the accumulation of electrostatic charge at the surface. Metal objects require little special preparation for SEM except for cleaning and mounting on a specimen stub. Nonconductive specimens tend to charge when scanned by the electron beam, and especially in secondary electron imaging mode, this causes scanning faults and other image artifacts. They are therefore usually coated with an ultrathin coating of electrically conducting material, deposited on the sample either by low-vacuum sputter coating or by high-vacuum evaporation. Conductive materials in current use for specimen coating include gold, gold/palladium alloy, platinum, osmium, iridium, tungsten, chromium, and graphite. BIOLOGICAL SAMPLES:- For SEM, a specimen is normally required to be completely dry, since the specimen chamber is at high vacuum. Hard, dry materials such as wood, bone, feathers, dried insects, or shells can be examined with little further treatment, but living cells and tissues and whole, soft-bodied organisms usually require chemical fixation to preserve and stabilize their structure. Fixation is usually performed by incubation in a solution of a buffered chemical fixative, such as glutaraldehyde, sometimes in combination with formaldehyde and other fixatives, and optionally followed by post-fixation with osmium tetroxide. The fixed tissue is then dehydrated. Because air-drying causes collapse and shrinkage, this is commonly achieved by replacement of water in the cells with organic solvents such as ethanol or acetone, and replacement of these solvents in turn with a transitional fluid such as liquid carbon dioxide by critical point drying. The carbon dioxide is finally removed while in a supercritical state, so that no gas-liquid interface is present within the sample during drying. The dry specimen is usually mounted on a specimen stub using an adhesive such as epoxy resin or electrically conductive double-sided

adhesive tape, and sputter-coated with gold or gold/palladium alloy before examination in the microscope

EFFECT OF SOME BASIC PARAMETERS

The quality of final image that is obtained can be altered by varying various parameters in a scanning electron microscope. In this section a brief description about these parameters and how the results are affected by varying them will be described. The parameters to be discussed here are:1. Acceleration voltage 2. Probe diameter 3. Probe current

EFFECT OF ACCELERATION VOLTAGE:- When theoretically considering the electron probe diameter alone, the higher the accelerating voltage, the smaller is the electron probe. However, there are some un-negligible demerits in increasing the accelerating voltage. They are mainly as follows: 1) Lack of detailed structures of specimen surfaces. 2) Remarkable edge effect. 3) Higher possibility of charge-up. 4) Higher possibility of specimen damage. In SEM, finer surface structure images can generally be obtained with lower accelerating voltages. At higher accelerating voltages, the beam penetration and diffusion area become larger, resulting in unnecessary signals (e.g., backscattered electrons) being generated from within the specimen. And these signals reduce the image contrast and veils fine surface structures. It is especially desirable to use low accelerating voltage for observation of lowconcentration substances.

EFFECT OF PROBE DIAMETER AND PROBE CURRENT In the SEM, the smaller the electron probe diameter on the specimen, the higher the magnification and resolution. However, the image smoothness, namely, the S/N ratio depends on the probe current. The probe current and the probe diameter are in the relation shown in Fig. 8. Namely, as the probe diameter is reduced, the probe current is reduced. It is therefore necessary to select a probe current suited for the magnification and observation conditions (accelerating voltage, specimen tilt, etc.) and the specimen.

APPLICATIONS

1. 2. 3. 4. 5. 6. 7. 8. 9.

Identification of metals and materials Particle contamination identification and elimination Classification of materials Product and process failure and defect analysis Examination of surface morphology (including stereo imaging) Analysis and identification of surface and airborne contamination Powder morphology, particle size and analysis Cleaning problems and chemical etching Welding and joining technology quality evaluation and failure investigation 10. Paint and coating failure and delamination investigation 11. Identification and elimination of corrosion and oxidisation problems 12. Contamination or stain investigation 13. Structural analysis 14. Reverse engineering of products and processes