Exercise 1

MICROSCOPY
PhBioSci 1
Maria Athena Monica M. Emnacen, RPh

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MICROSCOPE
• Micro (“small”), scope (“view”)
• instrument that magnifies objects too small to
be seen by the naked eye
 KINDS OF MICROSCOPE
• LIGHT MICROSCOPE
• ELECTRON MICROSCOPE
 KINDS OF MICROSCOPE
• LIGHT MICROSCOPE
– “light” refers to the process by which light
transmit the image to the eye.
– 2 basic types:
• DISSECTING MICROSCOPE
• COMPOUND MICROSCOPE
 KINDS OF MICROSCOPE
• LIGHT MICROSCOPE
• DISSECTING MICROSCOPE
– Permits viewing of opaque objects
– Most microscope can magnify up to 30x
PARTS OF DISSECTING MICROSCOPE

EYEPIECE
STAGE CLIP

STAGE

STAGE PLATE

ADJUSTING
KNOB
ARM
LIGHT
SOURCE

BASE
 KINDS OF MICROSCOPE
• LIGHT MICROSCOPE
• COMPOUND MICROSCOPE
– Optical instrument with a two-lens system
(objectives and eyepiece or ocular), controllable
illumination system and geared focusing
mechanism
– Requires that the material being examined be
sliced thinly for light to pass through
– Magnify the specimen up to 1500x
PARTS OF THE COMPOUND
MICROSCOPE
 PARTS OF THE COMPOUND
MICROSCOPE
• SUBDIVIDED into 3 PARTS:
– MECHANICAL PARTS
– MAGNIFYING PARTS
– ILLUMINATING PARTS
 PARTS OF THE COMPOUND
MICROSCOPE
• MECHANICAL PARTS:
1. BASE
- usually U- or V-Shaped
- Where the microscope firmly rests
2. ARM
- connects the base and the body tube together
- serves as a handle for the carrying of a microscope
3. STAGE
- the platform that holds/ supports the slide containing
the specimens
- has an opening in the center that allows light to pass
from below into the specimen for study
 PARTS OF THE COMPOUND
MICROSCOPE
• MECHANICAL PARTS:
4. PILLAR – the support or post of the base where the arm
is attached
5. STAGE CLIPS – hold the specimen firmly on the stage
6. SUBSTAGE – found below the stage
- holds the Abbe condenser above and the iris diaphragm
below
7. BODY TUBE – cylinder that attaches the dram tube into
the microscope
- acts as the passage way of light from the objective to
the eyepiece
 PARTS OF THE COMPOUND
MICROSCOPE
• MECHANICAL PARTS:
8. COURSE ADJUSTMENT – upper, larger knobs for the slow
movement of the body tube when focusing under the
low-power objective (LPO)
9. FINE ADJUSTMENT – for the slow movement of the body
when focusing under the high-power objective (HPO)
and other high power lenses.
10. REVOLVING NOSEPIECE – Carries the objectives
- turned to select the appropriate objective
- lenses must be “clicked” in place for the field to be
visible
11. DRAW TUBE – smaller cylinder attached to the upper
part of the body tube that holds the ocular
 PARTS OF THE COMPOUND
MICROSCOPE
• MAGNIFYING PARTS:
1. EYEPIECE or OCULAR – detachable cylinder located at
the upper end of the tube
- may have line inside that serves as a pointer and
rotates as the ocular is rotated.
2. OBJECTIVES – has two or three objectives attached to
the revolving nosepiece
– Scanner
– LPO
– HPO
– Oil immersion
 PARTS OF THE COMPOUND
MICROSCOPE
• MAGNIFYING PARTS:
2. OBJECTIVES
– Scanner
» Shortest cylinder with a very large opening
» Provided with a large lens for very low magnification
» Used to observe much wider field of the object
» Marked with “4x”
– Low-Power Objective (LPO)
» Shorter cylinder with a large lens opening
» Used to observe the general outline and locate various
parts of a specimen
» Focused using the course adjustment
» 10x magnification
 PARTS OF THE COMPOUND
MICROSCOPE
• MAGNIFYING PARTS:
2. OBJECTIVES
– High-Power Objective (HPO)
» Longer cylinder with a small lens opening
» Equipped with lenses of higher magnifying power and
used to study the detailed parts of a specimen.
» Focused by using the fine adjustment knob
» 40x magnification
– Oil Immersion Objective
» Longest cylinder with a very small lens opening
» Equipped with very small lenses for a very high
magnification and used to study details of specific parts
of the specimen
» A small drop of special oil is added so that the light
passing through it is uniformly distributed or is
dispersed.
 PARTS OF THE COMPOUND
MICROSCOPE
• ILLUMINATING PARTS:
1. MIRROR– Planar on one side and concave on the
other
- used to reflect light and lenses of the eye
- concave side of the mirror is used for the natural light
while the flat side used for artificial light
- held by a mirror rack
CAUTION: Never used direct sunlight
2. CONDENSER – Located on a substage held in place by
a rack.
- used to condense or concentrate the light reflected
from the mirror to the object or specimen being
examined.
 KINDS OF MICROSCOPE
• ELECTRON MICROSCOPE
– Can magnify very small details with a high resolution.
– High magnifying capacity is due to the use of elections
rather than the light to scatter off materials
– Magnifying at levels of more or less 500, 000x the
original size
– 4 basic types:
• TRANSMISSION ELECTRON MICROSCOPE
• SCANNING ELECTRON MICROSCOPE
• SCANNING TRANSMISSION ELECTRON MICROSCOPE
• SCANNING TUNNELING MICROSCOPE
 KINDS OF MICROSCOPE
• ELECTRON MICROSCOPE
• TRANSMISSION ELECTRON
MICROSCOPE (TEM)
– Uses high voltage electron beam
emitted by a cathode and
formed by magnetic lenses
– Using a very thin section of a
specimen, this electron beam
can carry and transmit
information about the internal
structure of a sample
– Magnify the specimen up to
more or less 200,000x
 KINDS OF MICROSCOPE
TRANSMISSION ELECTRON MICROSCOPE (TEM)
KINDS OF MICROSCOPE
• ELECTRON MICROSCOPE
• SCANNING ELECTRON MICROSCOPE (SEM)
– Make use of beam transmission
– Produces images by detecting secondary
electrons which are emitted from the surface
due to excitations by the primary electron
beam.
– The surface of a specimen is bombarded with
electrons to allow scientist to see a 3-
dimensional image of the specimen’s surface.
– Magnify 3, 000x to 10, 000x the original size
 KINDS OF MICROSCOPE
• ELECTRON MICROSCOPE
• SCANNING TRANSMISSION ELECTRON
MICROSCOPE (STEM)
– Specific type of TEM in which the electrons still pass
through the specimen
 KINDS OF MICROSCOPE
• ELECTRON MICROSCOPE
• SCANNING TUNNELING MICROSCOPE (STM)
– Newest machine that can reveal individual atoms
only 3 Angstrom across
– Works on the principle of electron tunneling
(tendency of electrons to jump between the tip of
a fine metal needle and the surface to be studied.
– Produces 3-dimensional image by scanning the
surface f the sample with a beam of electrons
 KINDS OF MICROSCOPE
• ELECTRON MICROSCOPE
• SCANNING TUNNELING MICROSCOPE