INTRODUCTION TO MICROSCOPY

THINK SMALL
 INTRODUCTION TO MICROSCOPY
• Our scopes are BRIGHT FIELD LIGHT
MICROSCOPE
• The scope uses visible light to create an
image, lenses that magnify the image, and the
image is displayed against a bright light
background. Microscopes magnify and
resolve a specimen.
• MAGNIFICATION: HOW BIG?
• **RESOLUTION: HOW CLEAR?
 Parts of the Microscope
HEAD

ARM
SLIDE CLIPS:
SECURES SLIDES

LIGHT
SOURCE BASE
 Parts of the Microscope

CONDENSER:
GATHERS LIGHT AND LIGHT
DIRECTS LIGHT ONTO
THE SPECIMEN SOURCE
 Parts of the Microscope

CONDENSER:
GATHERS LIGHT AND
DIRECTS LIGHT ONTO
THE SPECIMEN

IRIS DIAPHRAGM LIGHT

LEVER: SOURCE
CONTROLS LIGHT INTENSITY
 ADJUSTMENT KNOBS
Parts of the Microscope MOVE STAGE UP
AND DOWN

COARSE ADJUSTMENT
KNOB: MOVES STAGE UP
AND DOWN IN LARGE
INCREMENTS

FINE ADJUSTMENT KNOB:

STAGE ADJUSTMENT KNOBS: MOVES STAGE UP AND DOWN IN
MOVES THE STAGE RIGHT AND LEFT, SMALL INCREMENTS
FORWARD AND BACK.
 Parts of the Microscope
OCULARS:
Located on the head

REVOLVING NOSEPIECE:
HOLDS THE OBJECTIVE
LENSES
 A COMPOUND SCOPE: TWO SOURCES OF MAGNIFICATION
OR TWO LOCATIONS WHERE MAGNIFYING LENSES ARE FOUND

OCULARS
HOLDS SECOND SET
OF MAGNIFYING
LENSES

OBJECTIVE LENSES
FIRST SET OF MAGNIFYING LENSES,
 INTRODUCTION TO MICROSCOPY
• MAGNIFICATION OF A SPECIMEN is achieved with
MAGNIFYING LENSES
• MICROSCOPES WITH TWO SOURCES OF
MAGNIFYING LENSES ARE CALLED COMPOUND
MICROSCOPES
• NLC scopes are compound scopes.
• Two sources of magnification
– 1- objective lenses (4 types)
– 2- ocular lenses
 INTRODUCTION TO MICROSCOPY
• TOTAL MAGNIFICATION = TM

• TM ACHIEVED BY MULTIPLYING THE

MAGNIFICATION OF THE TWO SOURCES
OF MAGNIFYING LENSES
 INTRODUCTION TO MICROSCOPY
• TOTAL MAGNIFICATION = TM
TM = OBJECTIVE MAG. X OCULAR MAG. (10x)
• 4 OBJECTIVE LENSES available on our scopes:
– SCANNING LENS (4x),
– LOW POWER LENS (10x),
– HIGH POWER LENS (40x),
– OIL IMMERSION LENS (100x)
 INTRODUCTION TO MICROSCOPY
• TOTAL MAGNIFICATION (TM)
• Ocular X Objective = TM
• 10x x 4x = 40X
• 10X x 10x = 100x
• 10x x 40x = 400x
• 10x x 100x = 1000x
 INTRODUCTION TO MICROSCOPY
• RESOLUTION: HOW WELL CAN A MICROSCOPE
RESOLVE TWO POINTS OR SEPARATE TWO
STRUCTURES THAT ARE CLOSE TOGETHER

POOR RESOLUTION
Due to large light
wavelengths

GOOD RESOLUTION
SMALL WAVELEGTHS
The smaller the wavelength
the better the resolution.
 INTRODUCTION TO MICROSCOPY
• RESOLUTION WITH A LIGHT MICROSCOPE
CAN BE IMPROVED BY:
1. BLUE FILTER UNDER CONDENSER

BLUE LIGHT HAS

SMALL
WAVELENGHTS

2. USE OF OIL IMMERSION OBJECTIVE

 INSTRUCTIONS ON HOW TO USE A
MICROSCOPE
• CLEAN SCOPE
– Ocular, 40X and 100X objectives only,
• CLEAN PREPARED SLIDE
• Put 4X OBJECTIVE IN PLACE, POWER ON
• PLACE SLIDE ON STAGE AND HOLD WITH
MECHANICAL ARM (CLIPS)
• After setting slide on stage, bring stage all the
way up
 How to use a Microscope

SCANNING 4X
BRING STAGE DOWN OBJECTIVE IN
To place slid on stage PLACE

CONDENSER
ALL THE WAY UP
INSTRUCTIONS ON HOW TO USE A
MICROSCOPE
• MOVE STAGE ALL THE WAY UP
• 4x OBJECTIVE IN PLACE

• WHILE LOOKING THROUGH OCCULARS

• USE THE COARSE ADJUSTMENT KNOB TO MOVE THE
STAGE DOWN SLOWLY TO BRING SPECIMEN
TOWARD YOU AND IN FOCUS.
 How to use a Microscope

1 SCANNING 4X
OBJECTIVE IN
PLACE

STAGE UP
2

Move Coase
Adjustment Knob
CONDENSER DOWN slowly
ALL THE WAY UP
while looking through
oculars
INSTRUCTIONS ON HOW TO USE A
MICROSCOPE
• DO YOU SEE SOMETHING?
• FINE FOCUS WITH FINE ADJUSTMENT KNOB
until image is clear
• Center the specimen in the field of view.
• To make sure you are focused on the
specimen, MOVE SLIDE SIDEWAYS WITH STAGE
KNOBS. If image does not move when you
move the slide, then you are not focused on
specimen.
 How to use a Microscope
Use Fine
adjustment
knob to “fine
focus” image

Stage knobs are

use to move
slide sideways,
up, and down.
“Fine focus” a specimen means to
make the image sharp and clear.
 INSTRUCTIONS ON HOW TO USE A
MICROSCOPE
• Once FOCUSED ON SPECIMEN CLEARLY
• Center the specimen
• Switch to the next highest magnification
• USE FINE FOCUS AGAIN
• **Do not touch coarse adjustment knob
again. Only use with scanning (4x) lens.
INSTRUCTIONS ON HOW TO USE A MICROSCOPE

• REMEMBER: Do not touch the COARSE

ADJUSTMENT KNOB again after the initial
steps with the SCANNING 4X LENS.
• PARCENTRAL FEATURE OF THE MICROSCOPE
– HOLDS SPECIMEN ALMOST CENTERED IN THE FIELD
OF VIEW AS YOU CHANGE MAGNIFICATION.
• PARFOCAL FEATURE OF THE MICROSCOPE.
HOLDS SPECIMEN IN FOCUS AS YOU CHANGE
OBJECTIVES TO HIGHER MAGNIFICATION.
 INSTRUCTIONS ON HOW TO USE A MICROSCOPE
WORKING DISTANCE

• The WORKING DISTANCE IS:

– the distance between objective and slide.

As you increase magnification:

1.- ARE THE OBJECTIVES LONGER OR SHORTER?
2.- IS THE WORKING DISTANCE LARGER OR SMALLER?
• The Working Distance DECREASES as you
INCREASE MAGNIFICATION
 INSTRUCTIONS ON HOW TO USE A MICROSCOPE
FIELD OF VIEW

• The Diameter of the field of view refers to the

diameter of the circle of bright light against
which the specimen is seen.

• The Diameter of the field of view

– DECREASES as you INCREASE MAGNIFICATION
• ALWAYS MOVE SPECIMEN TO MIDDLE OF FIELD OF
VIEW BEFORE SWITCHING OBJECTIVES
 INSTRUCTIONS AFTER USING THE
COMPOUND LIGHT MICROSCOPE
• After using microscope:
• - Clean prepared slide, before placing back in
the correct slide tray
• - Clean Oculars, 40X, and then 100X lens with
lens paper & small amount of lens cleaner
• - Lower stage down,
• -Rotate the 4X objective in place
• - Wrap cord around arm, and store in
corresponding shelf with microscope number.