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Scanning Electron Microscope

Scanning Electron Microscope (SEM) scans electron beam over a surface to create an

image. The electrons in the beam interact with the sample producing various signals that can be

used to obtain information about the surface topography and composition

SEM’s consist of many components such as electron source, thermionic gun, field

emission gun, electromagnetic/ electrostatic lenses, vacuum chamber, sample chamber and stage,

computer, detectors, secondary electron detector (SED), backscatter detector, diffracted

backscatter detector (EBSD) and X ray detector (EDS). In addition, it requires a stable power

supply, vacuum and cooling system, vibration free space and need to be placed in an area that

isolates the instrument from ambient magnetic and electric fields.

In SEM imaging, it provides details surface information by tracing sample in a raster

pattern with an electron beam. The process begins with an electron gun generating a beam of

energetic electrons down the column and onto a series of electromagnetic lenses. The coils are

adjusted to focus the incident electron beam onto the sample. These adjustments cause

fluctuations in the voltage, increasing or decreasing the speed in which the electrons come in

contact with the specimen surface. It is controlled via computer and the operator can adjust the

beam to control magnification as well as determine the surface area to be scanned. The beam is

focused onto the stage where a solid sample is placed. Most samples require some preparation

before being placed in the vacuum chamber. The two most commonly used prior to SEM

analysis are sputter coating for non-conductive samples and dehydration of most biological

In addition, all samples need to be able to handle the low pressure inside the vacuum

chamber. The interaction between the incident electrons and the surface of the sample is

determined by the acceleration rate of incident electrons, which carry significant amounts of

kinetic energy before focused onto the sample. When the incident electrons come in contact with

the sample, energetic electrons are released from the surface of the sample. The scatter patterns

made by the interaction yields information on size, shape, texture and composition of the sample.

A variety of detectors are used to attract different types of scattered electrons which includes

secondary and backscattered electron as well as x rays.

Backscatter electrons are incidental electrons reflected backwards. The images provide

composition data related to element and compound detection. Although topographic information

can be obtained using a backscatter detection, it is not accurate as SED. Diffracted backscatter

electrons determine the crystalline structures as well as the orientation of minerals and micro-

fabrics. X rays emitted from beneath the sample surface can provide an element and mineral

formation. SEM produces black and white, three dimensional images. Image magnification can

be up to 10 nanometers and although it is not powerful as TEM counterpart, the intense

interactions takes place on the surface of the specimen provide a greater depth of view, higher

resolution and more detailed surface picture.

SEM’s have a variety of applications in a number of scientific and industry related fields

especially where characterizations of solid materials is beneficial. It provides information in

microstructures, analyze surface fractures, examine surface contaminations, reveal spatial

variations in chemical compositions, provide qualitative chemical analysis and identify

crystalline structures. It is essential applications in science, biology, gemology, medical, forensic

science, metallurgy and many more.

SEM’s advantages includes wide array of applications, detailed 3D, topographical

imaging, versatile information gathered from different detectors, easy to operate, works fast and

sample require minimal preparation actions.

Ahmed et. al (2013), used scanning electron microscope (SEM) for the surface

homogeneity and morphologies of α-Fe3O4/chitosan nanocomposite and chitosan electrode (See

Figure 1). Via this instrument, the α-Fe3O4 nanoparticles were observed (See Figure 1a). Also,

the α-Fe3O4 and chitosan nanocomposite can be seen and looks like a tiny particle hanging on it

(See Figure 1b) which shows the embedding and incorporation of the α-Fe3O4 nanoparticles in

chitosan film that represents the formation of α-Fe3O4/chitosan nanocomposite. The net shaped

may be attributed to electrostatic interactions between cationic chitosan and the surface charge α-

Fe3O4 nanoparticles. Thus, pure chitosan film changes the net morphology to homogenous and

regular form (See Figure 1c). In this case, α-Fe3O4 nanoparticles increases the

electroactivesurface area of the chitosan as the fold increases and change net which provides a

favorable environment for trapping heavy metals inside it.

Figure 1. SEM images of (a) α-Fe3O4 nanoparticles; (b) α-Fe3O4/chitosan nanocomposite; (c) Chitosan

[Preparation and Characterization of a Nanoparticles Modified Chitosan Sensor and Its

Application for the Determination of Heavy Metals from Different Aqueous Media]

Kim et. al (2010) demonstrated scanning electron microscope (SEM) for bacterial

cellulose/chitosan nanocomposite (See Figure 2). Images of freeze dried bacterial cellulose and

bacterial cellulose/chitosan nanocomposite were observed. It was reported that the mean

diameter of the nanofibrils was 40nm. Multiple layers structure can be seen from cross sectional

images with high aspect ratio and a well organized 3D network structure was determined. The

surface morphology of bacterial cellulose was changed after treating by chitosan. Due to the

presence of a thick layer of chitosan, bacterial cellulose nanofibrils was not observed and the

pore size becomes bigger. Chitosan molecules can penetrate into bacterial cellulose and forms

layers of bacterial cellulose/chitosan composite from cross sectional image. Lastly, bacterial

cellulose/ chitosan composite promotes cellular ingrowth when used as tissue engineering

Fig. 2 SEM images of bacterial cellulose (a. surface morphology, c. cross section morphology) and chitosan and

bacterial cellulose composite (b. surface morphology, d. cross section morphology)

[Preparation and Characterization of Bacterial Cellulose and Chitosan Composite for Potential

Buiomedical Application]

Rahmi et. al (2017) analyzed the texture and morphology of materials with the use of

scanning electron microscope (SEM) (See Figure 3). The results showed chitosan film smooth

structure (See Figure 3a) and the cellulose particles extracted from oil palm empty fruit bunch

has an average particle size with a range of 5 to 15 µm (See Figure 3b). The image of the surface

of chitosan/cellulose composite film were observed to be homogenous (See Figure 3c). This

homogenous structure attributed to the reinforcement of composite film but the surface became

rougher when the cellulose particles were added.

Figure 3. SEM analysis of (a) chitosan film (b) cellulose particles and (c) chitosan and cellulose composite film.

[Preparation of Chitosan Composite Film Reinforced with Cellulose Isolated from il Plam Empty

Fruit Bunch and Application in Cadmium ions Removal from Aqueous Solutions]

Lin et. al (2012) metalized the surface of the chitosan/cellulose blend films (CS/BC) with

gold through sputtering within 20 s. All of the films showed a homogenous structure without the

separation of the phase between chitosan (CS) and cellulose blend films (BS). A porous structure

and a well organized network were also observed. It was said that the pore formation was caused

through the leaching of Zn(OH)2 precipitate while washing. The mechanism can be described via

the interaction of or zinc ions with hydroxide and precipitate in the solution. Also, the precipitate

is leached during washing process in tap water and deionized water which resulted in pore in

films. Blending with chitosan, chitosan/cellulose blend film (CS/BS) becomes denser than RBC

film. Also, the densities of chitosan/cellulose blend film (CS/BS) increases as the chitosan

content increases.
Figure 4. SEM images of RBC and blend films.

[Preparation and Characterization of Chitosan/Cellulose Blend Films Using ZnCl2∙3H2O as a