Bio 22 - General Zoology

MTh 7-10AM

Rohani B. Cena, DVM, MSc

Rm108 Institute of Molecular Biology and
Biotechnology, National Institutes of Health, UP
Manila
MWTh 8-10AM
MICROSCOPY
Types of Microscope
Simple Light Microscope (Uses single lens)

Compound Microscope (Uses a set of lenses)

Stereoscopic/Dissecting Microscope
Fluorescence Microscope
Phase-contrast Microscope
Differential Interference Microscope

Electron Microscope(Uses electron beams on electromagnetic lenses)

Scanning Electron Microscope (SEM)
Transmission Electron Microscope(TEM)
Simple Light Microscope
Compound Light Microscope
Stereoscopic/Dissecting Microscope

Sample View
Fluorescence Microscope

Sample View
Phase Contrast Microscope

Sample View
Differential Interference Microscope

Sample View
Scanning Electron Microscope

Sample View
Transmission Electron Microscope

Sample View
Mechanical Parts

- Base -Nosepiece
- Pillar -Dust shield
- Inclination Joint -Coarse Adjustment Knob
- Arm -Fine Adjustment Knob
- Stage -Condenser Adjustment knob
- Draw tube -Iris Diaphragm Lever
Optical Parts
Terms to Know
Magnifying Power Numerical Aperture
10x, 40x, 100x
0.30, 0.65, 1.30
-ability to magnify the
- Light refracting capability
view of an object

Resolving Power
-ability to reveal adjacent details as separate and distinct.
-typical compound microscope has a resolving power of 0.2-0.4 nm

Working Distance
- distance between the lens of the objective and the object on
the slide when it is in focus.
 Differences between the Old and New Models

Old New
Objectives (3) LPO, HPO, OIO (4) Scanning, LPO,
HPO and OIO
Not spring-loaded Spring-loaded
Adjustment Separate Joined
Knobs
Illumination Ambient Light Light Bulb
(Light Source) using Mirror
Stage Clips Immovable Movable
 Differences between Compound Microscope and
Stereoscope/Scanning Microscope

Compound Stereoscope
Magnification 10x-100x 4x

Orientation Separate Joined

Movement of object Inverted Same

across field
Type of Image Virtual Real
Produced
Formulas
Computing for the Linear Magnification
Magnifying Power x Magnifying Power
of Objective of Ocular Lens

Example:
Magnifying Power of HPO = 40x
Magnifying power of Ocular =10x

40 x 10= 400x
When using the Ocular micrometer:

Calibration Factor must first be computed,

Number of Lines in Stage Micrometer x 0.01mm
Number of Lines in Ocular Micrometer
Take note: Lines in the ocular are usually smaller than in the stage
Example:
Number of Lines in stage = 5 Ocular

Stage
Number of Lines in ocular = 9

5/9 x 0.01mm = 0.0055mm = length per space in

the ocular micrometer
Computing for the
Actual Size of the Object
Actual Size:
Number of spaces in x Calibration
Ocular Micrometer Factor

Example:
Number of spaces in ocular micrometer = 12
Computed Calibration Factor = 0.0055mm

12 x 0.0055mm = 0.0648mm = Actual size of object (mm)

In nanometers:
0.0648 mm x 100 nm = 6.48 nm = Actual size of object (nm)
1mm
 Computing for the Magnification of
the Illustration
Size of Drawing / Actual Size of Object

Example:
Size of Drawing = 3 cm or 30mm
3cm
Actual size of object = 0.0648 mm

30mm/0.0648mm = 462.96x
or 463x