ELECTRON MICROSCOPY

Simplified Study Notes | SEM & TEM

Forensic Science • Paper 4 • Module 33

1. INTRODUCTION TO ELECTRON MICROSCOPY

Electron Microscopes use a beam of high-energy electrons instead of light to examine objects at very high
magnification. They can reveal topography (surface shape), morphology (form and size), composition
(elements present) and crystallographic information.

Why EM? Light microscopes are limited to ~500–1000× magnification and 0.2 µm
resolution. EM improves resolution by 1000× using electrons (shorter
wavelength).

History TEM – developed by Max Knoll & Ernst Ruska (Germany, 1931). SEM – first
built 1942, first commercial model 1965.

Types Microscopes are of three types: Optical, Charged-particle (electron/ion), and

Scanning probe.

■ Key Points to Remember

• EM uses electromagnets as lenses (not glass).
• Shorter wavelength of electrons → much higher resolution than light.
• TEM = 2D cross-section image | SEM = 3D surface image.
• Ernst Ruska won the Nobel Prize in Physics in 1986 for developing TEM.

2. PRINCIPLE OF ELECTRON MICROSCOPY

Electrons accelerated to a fixed voltage all have the same wavelength → they form a monochromatic beam.
When this beam hits the specimen, three types of interactions occur:

Interaction What happens? Used in

Unscattered Electron passes straight through

TEM – bright areas
transmission (thin region)

Direction changes, NO energy loss

Elastic scattering BSE imaging in SEM
(backscattered if >90°)

Energy transferred to specimen; Secondary electrons (SE)

Inelastic scattering
electron slows down in SEM

Signals Generated (important for 4-mark answer):

• Backscattered Electrons (BSE): Electrons bounce back at >90°. Higher atomic-number elements produce
MORE BSEs → appear brighter. Used for composition contrast in SEM.
• Secondary Electrons (SE): Outer-shell electrons ejected with low energy (<50 eV). Come from surface (<2
nm deep). Used for surface topography in SEM.
• Characteristic X-rays: When inner-shell electron is ejected, a higher-shell electron fills the gap → emits X-ray
with element-specific energy. Used for elemental analysis (EDX/EDS).
• Auger Electrons: Alternative to X-ray emission. Energy released when inner-shell vacancy is filled is
transferred to another electron, ejecting it. Used for surface chemistry analysis.
• Cathodoluminescence: Electrons promote valence electrons to conduction band → emits UV/Visible/IR light
when they return. Used in mineralogy and semiconductor studies.

■ Signal Memory Tips

• BSE → atomic number contrast | SE → surface topography.
• X-rays from inner-shell ionization are 'characteristic' → tell us WHICH element is present.
• Auger electrons = alternative way atom stabilizes after inner-shell ionization.
 3. SCANNING ELECTRON MICROSCOPE (SEM)

SEM scans a focused electron beam over the surface of the specimen and detects signals (SE & BSE) to build a
3D-looking surface image.

Fig 1a – SEM Schematic Diagram (with labels)

SCANNING ELECTRON MICROSCOPE (SEM) – Schematic

Acc. Voltage:
2–50 kV Electron Gun
(Tungsten/LaB■)

Anode

Condenser Lens

e■ Beam
Scanning Coils (X & Y deflection)

Objective Lens

SPECIMEN
Backscattered Secondary
(on Stage)
Electron Detector Electron Detector

■ High Vacuum 3D IMAGE

Column (10■■–10■■ Torr)
CRT Monitor

Fig 1b – SEM Block Diagram (signal flow)

SEM – Block Diagram (Signal Flow)

Electron Condenser Scanning Objective Specimen SE / BSE CRT

Gun Lens Coils Lens (Surface) Detector Display

e■ emitted beam condensed beam deflected focused spot signals generated signals detected image formed

How SEM Works – Step by Step:

Step 1: Electron Source Tungsten filament or LaB■ cathode inside electron gun generates
electrons.

Step 2: Acceleration Electrons accelerated at low voltage (2–50 kV).

 Step 3: Condenser Lens Narrows beam to a fine spot (~0.5–5 nm diameter).

Step 4: Scanning Coils Deflect beam in X and Y directions to scan across specimen surface.

Step 5: Objective Lens Focuses the fine beam onto the specimen.

Step 6: Signal Detection SE detector (Everhart-Thornley) picks up secondary electrons for

surface images. BSE detector picks up backscattered electrons for
composition images.

Step 7: Image Formation Signals displayed as varying brightness on a CRT/monitor → 3D

surface image.

■ SEM Quick Notes

• SEM = SURFACE imaging → 3D looking image.
• Low voltage (2–50 kV) + moderate vacuum (10■■ to 10■■ Torr).
• Two detectors: SE detector (topography) + BSE detector (composition/atomic number).
• Non-conducting samples must be coated with gold/carbon before SEM.
• Condenser lens → narrows beam | Objective lens → focuses beam on specimen.
 4. TRANSMISSION ELECTRON MICROSCOPE (TEM)

TEM passes a high-energy electron beam through an ultra-thin specimen. The transmitted electrons are
magnified by a series of lenses to form a 2D high-resolution image on a fluorescent screen or CCD camera.

Fig 2a – TEM Schematic Diagram (with labels)

TRANSMISSION ELECTRON MICROSCOPE (TEM) – Schematic

HV Supply: Electron Gun

200–300 kV (Thermionic / FEG)

Anode

Condenser Lens

SPECIMEN
(Ultra-thin, ~100 nm)

Objective Lens

High Vacuum
10■■–10■■ Torr
Intermediate Lens

Projector Lens

Fluorescent Screen
Output: / CCD Camera
2D high-res image

Fig 2b – TEM Block Diagram (signal flow)

TEM – Block Diagram (Signal Flow)

Electron Condenser Thin Objective Intermediate Projector Screen /

Gun Lens Specimen Lens Lens Lens CCD

e■ emitted beam collimated electrons pass image formed magnified projected 2D image
through recorded

TEM Components – What Each Part Does:

 Electron Gun Thermionic emission from tungsten filament (heated to ~2700 K) or LaB■
(Illumination system)
source. High voltage supply of 200–300 kV accelerates electrons to high
kinetic energy. Only ~1% of electrons pass through the anode hole.

Condenser Lens Converges the beam into a near-parallel spot incident on the specimen.
Controls the beam diameter and intensity on specimen.

Specimen Stage Ultra-thin specimen (~100 nm) placed on a copper grid. Loaded through
airlock (to maintain vacuum). Can tilt and move in 3 axes so all regions can
be examined.

Objective Lens Electromagnetic lens (high-excitation current coil). Produces initial

magnified image. Most critical lens – determines resolution. Needs highly
stable current supply. Generates a lot of heat → insulated with thermal
distributors.

Intermediate Lens + Intermediate lens selects whether to see an image or diffraction pattern.
Projector Lens
Projector lens forms a large final image on the fluorescent screen (few cm
diameter). Short focal length minimizes distortion from high-angle
scattering.

Screen + Detectors + Fluorescent screen made of Zinc Sulfide (ZnS) converts electron
Image Recorder
diffraction into visible light. CCD camera or photographic film records the
final image. Lighter areas = thinner regions (more electrons pass). Darker
areas = denser regions (fewer electrons pass).

■ TEM Quick Notes

• TEM = TRANSMISSION imaging → 2D internal/cross-section image.
• High voltage (200–300 kV) + high vacuum (10■■ to 10■■ Torr).
• Specimen must be ultra-thin (~100 nm) for electrons to pass through.
• Light areas = thin/less dense | Dark areas = thick/more dense.
• TEM can also produce electron diffraction patterns for crystallography.
• Magnification range: 10² to 10■ × (can image individual atoms).
 5. SEM vs TEM – Quick Comparison Table

Feature SEM TEM

Image Type 3D surface image 2D cross-section image

Specimen Thick / bulk (coated) Ultra-thin (~100 nm)

Electrons used Secondary / Backscattered Transmitted electrons

Magnification Up to ~500,000× Up to 1,000,000×

Resolution ~1–20 nm ~0.1–0.2 nm

Voltage 2–50 kV 100–300 kV

Vacuum Moderate (10■■–10■■ Torr) High (10■■–10■■ Torr)

GSR, surface morphology, Internal cell structure,

Forensic use
finger print, fibers nano-particles, drugs

Developed 1942 (commercial 1965) 1931 (Knoll & Ruska)

6. FORENSIC APPLICATIONS OF ELECTRON MICROSCOPY

Every crime leaves traces behind. Microscopic examination of these traces provides critical evidence. Key
forensic applications include:

■ Gunshot Residue (GSR) SEM-EDX detects Lead (Pb), Antimony (Sb), Barium (Ba) particles
Analysis
on shooter's hands/clothing. Particle morphology (spherical shape,
specific size) confirms GSR.

■ Firearms Identification SEM compares striation marks on bullets/cartridge cases with

test-fired rounds. Unique marks left by firearm barrel/firing pin help
match weapon to crime.

■ Gemstone & Jewellery SEM reveals surface features, inclusions and treatment marks.
Investigation
EDX identifies elemental composition to distinguish natural from
synthetic gems.

■ Paint & Fiber Examination SEM-EDX analyzes paint flakes from hit-and-run accidents –
identifies vehicle make/model. Fiber cross-sections compared to
known samples for source identification.

✍■ Handwriting & Document SEM examines ink particles, paper fibers and toner deposits to
Forgery
detect forgery, altered documents or counterfeit currency at micro
level.

■ Drug Analysis TEM used to study nanostructure of drug particles; SEM examines
morphology of drug crystals to identify substance and
manufacturing source.

■■ Trace Evidence Comparison Glass fragments, soil particles, hair cross-sections and explosive
residues examined for morphological and compositional
comparison between suspect and crime scene samples.
 ■ Filament Bulb Investigation SEM examines whether a filament bulb (e.g., at traffic signal) was
ON or OFF at time of accident based on surface oxidation patterns
and hot-stretch deformation.

■ Forensic Application Tips

• SEM-EDX is the GOLD STANDARD for GSR analysis in forensic labs.
• SEM gives 3D morphology → ideal for surface evidence (paint, GSR, fibers).
• TEM gives internal structure → ideal for drugs, biological samples, nano-particles.
• Both instruments non-destructive (mostly) → evidence preserved for court.

7. LIMITATIONS

Limitation SEM TEM

Sample prep
Non-conductors need metal coating (gold/carbonVery
sputter
tedious
coating)
– specimen must be thinned to ~100 nm

Vacuum needed Moderate vacuum required High vacuum required

Image type Only surface – cannot see internal structure Only 2D projection – no 3D surface info

Sample size Can handle larger samples Only tiny specimens fit (3 mm grid)

Cost Expensive but less than TEM Very expensive + complex maintenance

Live samples Not possible (vacuum kills live cells) Not possible

8. REVISION SUMMARY (4–5 Mark Answer Guide)

SEM – 4-5 Mark Answer Points TEM – 4-5 Mark Answer Points
• Definition: SEM scans a focused electron • Definition: TEM transmits electrons through
beam over specimen surface to produce 3D ultra-thin specimen to form 2D
surface images. high-resolution images.

• Electron source: Tungsten filament / LaB■ • Electron source: Thermionic gun (tungsten
cathode; acceleration voltage 2–50 kV. filament/LaB■); voltage 200–300 kV.

• Lenses: Condenser lens (narrows beam) → • Specimen must be ultra-thin (~100 nm);
Scanning coils (X-Y deflection) → Objective placed on copper grid; loaded via airlock.
lens (focuses).

• Signals used: Secondary Electrons (SE) for • Components: Electron gun → Condenser →
topography; Backscattered Electrons (BSE) Specimen stage → Objective → Intermediate
for composition. → Projector.

• Detectors: Everhart-Thornley detector (SE) + • Image formation: Light areas = thin regions;
solid-state detector (BSE). dark areas = dense regions.

• Output: 3D surface image on CRT; vacuum: • Screen: ZnS (Zinc Sulfide) fluorescent screen
10■■ to 10■■ Torr. converts electrons to visible light; CCD
records image.

• Forensic use: GSR analysis, paint/fiber • Magnification: 10² to 10■ ×; can resolve
examination, counterfeit currency, firearms atomic columns; also produces electron
identification. diffraction patterns.
 • Forensic use: Nanoparticle analysis, drug
morphology, biological trace evidence.

■ Final Memory Tricks

TRICK: SEM → 'S' for Surface (3D). TEM → 'T' for Through / Thin (2D).
SEM voltage LOW (2–50 kV) | TEM voltage HIGH (200–300 kV).
SEM specimen = thick + coated | TEM specimen = ultra-thin (~100 nm).
Both use: Electron gun → Electromagnetic lenses → Detector/Screen → Image.
Forensic: SEM-EDX = GSR gold standard. TEM = internal nanostructure analysis.