,

SCANNING ELECTRON MICROSCOPY.

SUBJECT,

ANALYTICAL CHEMISTRY

INTRODUCTION
The classical method for obtaining detailed information about the physical nature of surfaces was optical microscopy, which is still an important technique. The resolution of optical microscopy is limited by diffraction effects; however, to about the wavelength of light .Much higher resolution information is obtained by using one of the electron microscopic methods. The two most important methods are scanning electron microscopy and transmission electron microscopy. Although, scanning and transmission electron microscopy methods have many similarities, scanning electron microscopy can be thought of as providing images of external morphology, similar to those accessed by the human eye. In contrast, transmission electron microscopy probes the internal structure of solids and gives us information about micro structural details that is not familiar to the eye. We will strictly restrict our discussion here to scanning electron microscopy.

In obtaining an SEM image, a finely focused beam of electrons is scanned across the sample in raster scan by scan coils. The resulting raster scanning pattern is similar to that used in a cathode ray tube of television set in which the electron beam is 1)swept across the surface linearly in the x- direction ,2) returned to its starting position ,and 3)shifted downward in the y-direction by a standard increment .This process is repeated until a desire area of the surface has been scanned .In more recent instruments ,the same effect is achieved by digital control over the beam position on the sample .In either the analog scanning case or in digital systems, a signal is received above the surface and stored in computer where it is ultimately converted to an image . Several types of signals are produced from a surface in this process ,including backscattered ,secondary and Auger electrons-ray fluorescence photons ;and other photons of various energies .All of these processes have been used for these studied. In SEM instruments, backscattered and secondary electrons are detected and use to construct an image.
Electron gun and optics:-

The electron source is usually a tungsten filament source, although field emission guns are also employed for high resolution work. The electrons are accelerated to energy between 1 and 30 keV. The magnetic condenser and objective lenses systems reduce the spot size to a diameter of 2-10 nm when it reaches specimen .The condenser lens system ,which consist of one or more lenses, is responsible for the throughput of the electron beam reaching the objective lens; the objective lens determines the size of the beam hitting the sample surface .

Scanning with an SEM is accomplished by the two pairs of electromagnetic coils located within the objective lenses; one pair deflects the beam in x-direction across the sample and the other pair deflects it in y-direction .Scanning is controlled by applying an electrical signal to one pair of scan coils such that the electron beam strikes the sample to one side of the center axis of the lens system. By varying the electrical signal to this pair of coils as a function of time, the electron beam is moved in straight line across the sample and then returned to its original position .After completion of the line scan, the other set of coils is used to deflect the beam slightly, and the scanning of the beam using the x coils is repeated .Thus, by rapidly moving the beam the entire surface can be irradiated with the electron beam .The signals to the scan coils can be either analog or digital. Digital scanning has the advantage of vary reproducible movement and location of the electron beam .The signal from the sample can be encoded and stored in digital form analog with digital representations of the x and y positions of the beam. In analog SEMs, the signals that drive the electron beam in the x and y directions also drive the horizontal and vertical scan of a CRT. The image of the sample is produced by using the output of a detector to control the intensity of spot on the CRT. Thus this method of scanning produces a map of the sample in which there is one to one correlation between the signal produced at a particular location on the sample surface

and a corresponding point on the CRT. The magnification is achievable in SEM image by following relation,
M= W/w

Where, W is the width of the CRT display and w is the width of the single line scan across the sample .Because W is constant, increased magnification is achieved by decreasing w.
Samples and sample Holders:

. Sample chambers are designed for rapid changing of sample. The sample holder, or stage, in most instruments is capable of holding samples of many centimeters on an edge .Furthermore, the stage can be moved in x, and y, z directions, and it can be rotated about each axis .As consequences, the surfaces of most samples can be viewed from almost any perspective.
Electron Beam Interactions-

The versatility of the SEM and electron microprobe for the study of solids arise from the wide variety of signals generated when the electron beam interact with the solid. We consider just three of these signals: backscattered electrons ,secondary electrons ,and x-ray emission .The interactions of a solid with an electron beam can be divided into two categories:elestic interaction that effect trajectories of the electrons in the beam without altering the energies significantly and inelastic interaction ,which result in transfer of part or all of the energy of the electron to the solid .The excited solids then emits secondary electrons, Auger electrons ,x-rays ,sometimes longer wavelengths photons.

When an electron collides elastically with an atom, the direction of the electron changes, but the speed of the electrons is virtually unaffected so that kinetic energy remain essentially constant .The angle of deflection for any given collision is random and can vary from 0 to 180 The energy of the beam is assumed to be 20 keV, which is typical. Note that such a beam penetrates to a depth of 1.5mor more .Some of the electrons eventually lose energy by inelesic collision and remain in the solid ;the majority .however undergo numerous collision and as a result ,eventually exit from the surface as backscattered electrons. It is important to note that the beam of backscattered electrons has much larger diameter than the incident beam that is for a 5m incident beam, the backscattered beam may have diameter of several micrometer. The diameter of backscattered beam is one of the factors limiting the resolution of an electron microscope. Backscattered electrons have a broad energy spread, ranging from 50eV up to the energy of the incident beam. When the surface of a solid bombarded with an electron beam having an energy of several keV, electrons having energy of 50eV or less are emitted from the surface along with the backscattered electrons. The number of the secondary electrons is generally one half to one fifth or less the number of backscattered electrons .Secondary electrons are produced as a result of interactions between energetic beam electrons and weakly bound conduction electrons in the solid ,which leads to ejection of the conduction band electrons with a few electrons volt of energy. Secondary electrons are produced from a depth of only 50 to 500 A and exit in a beam that is slightly larger in diameter than the incident beam .Secondary electrons can be prevented from reaching the detector by applying the small negative bias to the transducer housing. The region the electrons penetrate is known as the interaction volume .Even though radiation is generated within this volume, it will not be detected unless it

escapes from the specimen-rays is not easily absorbed and so most escapes. For x- rays the sampling volume, which is the volume of material contributing to the x- ray signal, is on the order of the interaction volume .The backscattered electrons will not escape if they have penetrated more than a fraction of a micrometer .Thus the backscattered signal originates from a much smaller volume .The secondary electron signal arises from a region that is on the order of diameter of the incident electron beam. For this reason, secondary electrons are capable of giving much higher resolution than the other signals, and they are the most widely used signals in the SEM system

Secondary electron detectors are standard equipment in all SEMs, but it is rare that a single machine would have detectors for all possible signals. The signals result from interactions of the electron beam with atoms at or near the surface of the sample. In the most common or standard detection mode, secondary electron imaging or SEI, the SEM can produce very high-resolution images of a sample surface, revealing details less than 1 nm in size. Due to the very narrow electron beam, SEM micrographs have a large depth of field yielding a characteristic three-dimensional appearance useful for understanding the surface structure of a sample. Back-scattered electrons (BSE) are beam electrons that are reflected from the sample by elastic scattering. BSE are often used in analytical SEM along with the spectra made from the characteristic X-rays, because the intensity of the BSE signal is strongly related to the atomic number (Z) of the specimen. BSE images can provide information about the distribution of different elements in the sample. For the same reason, BSE imaging can image colloidal gold immuno-labels of 5 or 10 nm diameters, which would otherwise be difficult or impossible to detect in secondary electron images

in biological specimens. Characteristic X-rays are emitted when the electron beam removes an inner shell electron from the sample, causing a higher-energy electron to fill the shell and release energy. These characteristic X-rays are used to identify the composition and measure the abundance of elements in the sample.

Sample preparation:-

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,
The Environmental SEM

The electron beam deposits considerable charge in the sample .In conductive materials, the charge flows through the sample stage to the ground .In insulating materials, the charge accumulated, causing local variation in the secondary electron emission and deflecting the electron beam itself. 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 . Unfortunately, most biological specimens and most mineral samples do not conduct. variety of techniques have been developed for obtaining for SEM images of non conducting samples, but the most common approaches involve coating the surface of the sample with a thin metallic film produced by sputtering or by vacuum evaporation.

Nonconducting specimens may be imaged uncoated using environmental SEM (ESEM) or low-voltage mode of SEM operation. Environmental SEM instruments place the specimen in a relatively high-pressure chamber where the working distance is short and the electron optical column is differentially pumped to keep vacuum adequately low at the electron gun. The high-pressure region around the sample in the ESEM neutralizes charge and provides an amplification of the secondary electron signal. Low-voltage SEM is typically conducted in an FEG-SEM because the field emission guns (FEG) is capable of producing high primary electron brightness and small spot size even at low accelerating potentials. Operating conditions to prevent charging of nonconductive specimens must be adjusted such that the incoming beam current was equal to sum of out coming secondary and backscattered electrons currents. It usually occurs at accelerating voltages of 0.34 kV.
APPLICATION;-

1) Scanning electron microscopy provides morphological and topographic information about a wide variety of solid surfaces.

2) Identification of metals and materials.

3) Paint and coating failure and delaminating investigation. 4) Examination of biological samples.

5) Particle contamination identification and elimination. REFERENCES: 1) DOUGLAS A. SKOOG (2007) , INSTRUMENTAL ANALYSIS. 2) WIKIPEDIA .