MIC 125 | Nurul Insyirah Sedek

CHAPTER 3 : MICROSCOPY
LIGHT PROPERTIES
Wavelength: distance between 2 identical peaks, represent repeating pattern of travelling
energy

STUDENT MICROSCOPE

1. Magnification – ability to
enlarge object
 The objective lens forms the
magnified real image
(micrograph)
 Real image is projected to
eyepieces and magnified to form
virtual image
 Total magnification = power of
objective × power of eyepieces
 Depending on the size and
curvature of the lens, the image
appears enlarged
2. Resolving power – ability to
show detail
3. Depth of focus -thickness of a specimen that can be seen at one time
 Magnification ↑ , depth of focus ↓
4. Field of vision – surface area of view. Area↓ , magnification ↑
 MIC 125 | Nurul Insyirah Sedek

Eyepiece – allows you to view the specimen Stage clips – hold microscope slide on stage
Objective lens – collect light coming through Adjustment knob – moves stage up and down
the object and magnifies it to focus on specimen
Nose piece – hold objective lens in place Light source – shine light up through the
slide and directed to mirror
Stage – hold and support microscopes slides Base – prevents the microscope from tipping
over
Condenser – controls amount of light

TYPE OF LIGHT MICROSCOPES

▪ Bright-field ▪ phase-contrast
- Most widely used - transforms subtle changes in
- Specimen is darker > light waves into differences light
surrounding field intensity
- Live and stained specimen - best for observing intracellular
▪ Dark-field (inside a cell) structures
- Brightly illuminated specimens - have microscope with special
surrounded by darker field condenser
- live and unstained specimen

▪ Differential interference contrast ▪ Fluorescence microscope

(DIC)
- Nomarski interference contrast
(NIC)
- Used to enhance the contrast in
unstained, transparent specimen
- Produce 3D image, >lively
- Higher resolution
- Modified compound microscope
w UV radiation source and filter
that protect viewer’s eyes
- Uses dyes that produce visible
light when bombarded w shorter
UV rays – make it fluorescence
- Useful to diagnose infections
(hospital, lab)
 MIC 125 | Nurul Insyirah Sedek

▪ Confocal microscope ▪ Digital microscopy

- Fluorescence or unstained - Uses computer tech. to
specimen images combined to automatically focus, adjust light
form 3D image & take photo of specimens
- Use laser light to obtain thin - Can be directly upload & viewed
section of specimens (internal) online

▪ Electron microscope ii. Transmission electron

- Uses beam of electrons , travel in microscopes (TEM)
wavelike patterns - Transmit electron THROUGH
- Electron waves 100 000x shorter the specimen (dalam specimen)
than visible light, tremendous - Darker = thicker/denser , lighter
power to resolve vv small = transparent/less dense
structures ▪ Stereo microscope /dissecting
- Magnification between 5 000x to microscope
1 000 000x - No mechanical stage, objective
i. Scanning electron lens
microscopes (SEM) - Observe large specimen, low
- Use to observe SURFACE of the magnification
specimen - Using light reflected from
- Provide 3D view surface of specimen