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
10x, 40x, 100x
-ability to magnify the
view of an object

Numerical Aperture
0.30, 0.65, 1.30
- Light refracting capability

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
Knobs
Illumination
(Light Source)
Stage Clips

Separate

Joined

Ambient Light
using Mirror
Immovable

Light Bulb
Movable

Differences between Compound Microscope and

Stereoscope/Scanning Microscope

Compound

Stereoscope

Magnification

10x-100x

4x

Orientation

Separate

Joined

Movement of object
across field

Inverted

Same

Type of Image
Produced

Virtual

Real

Formulas

Computing for the Linear Magnification

Magnifying Power
of Objective

Magnifying Power
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
Number of Lines in ocular = 9

Ocular
Stage

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
1mm

= 6.48 nm = Actual size of object (nm)

Computing for the Magnification of

the Illustration
Size of Drawing

/ Actual Size of Object

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

30mm/0.0648mm = 462.96x
or 463x

3cm