Lab 5

Microscopy
Lab Manual pages 71-85
 What is Microscopy?
• Microscopy: the technical field of using
microscopes to view samples or objects.
Very important tool in science
which led to many scientific
discoveries.
•Anton van Leeuwenhoek, 16th
century viewer of bacteria
•Robert Hooke, first person to
coin the term “cell”
 Types of Microscopes
• Optical/Light
Microscopes, which
include compound and
dissecting/stereo
microscopes

• Uses light and lenses to

magnify images
Dissecting Compound

• Simple vs. Compound

Scopes
Types of Microscopes
• Non-optical microscope
• Scanning Electron
Microscope
(SEM)/Transmission
Electron Microscope (TEM)
• Produce images by
scanning the sample with a
high energy beam of
electrons
 How They Work
Optical
Microscope Non-optical
 Optical vs. Non-optical
Characteristics
  Optical Non-optical
Magnification 40-1200X 25-250,000X

Possible Samples Can be living or dead Must be trimmed and

coated/fixed

Resolution Moderate Very High

Benefits Less expensive Very wide depth of

field (3D Images)

Limitations Limited Depth of field Very Expensive and

more difficult to
operate

Depth of field: vertical distance that remains in focus at one time

(decreases as magnification increases in light microscopes)
 Parts of the Light Microscope
Ocular

Eyepiece,
closest piece to
the eye
 Parts of the Light Microscope
Ocular
Objective

The lens or
mirror of a
microscope
 Parts of the Light Microscope
Ocular
Objective

Stage

Platform where
slides are
placed (can be
adjusted up or
down)
 Parts of the Light Microscope
Ocular
Objective

Stage

Stage Controls

Holds the slides

in place for
viewing
 Parts of the Light Microscope
Ocular
Objective

Stage

Stage Controls
Condenser
Lens that
focuses light on
the specimen
 Parts of the Light Microscope
Ocular
Objective

Stage

Stage Controls
Condenser
Function is to stop
the flow of light Diaphragm
(can be adjusted
left or right)
 Parts of the Light Microscope
Ocular
Objective

Stage

Stage Controls
Condenser
Controls the
strength of the Diaphragm
light being used Light Intensity Control
 Parts of the Light Microscope
Ocular
Objective

Stage
Coarse
Stage Controls Focus Knob
Condenser
Fine
Used to focus in Focus Knob
on specific
Diaphragm
features of the Light Intensity Control
sample
 Parts of the Light Microscope
Ocular
Objective
Power
Stage On/Off
Coarse
Stage Controls Focus Knob
Condenser
Fine
Used to power Focus Knob
the microscope
Diaphragm
on and off Light Intensity Control
 Magnification
Magnify/Magnificaton: to enlarge something in
appearance.
3 Objectives:
1. Scanning (4X)
2. Low (10X)
3. High (40X)
The ocular lens (eyepiece) is 10X
Total magnification= objective x ocular
 Magnification
Magnify/Magnificaton: to enlarge something in
appearance.
3 Objectives:
1. Scanning (4X)
2. Low (10X)
3. High (40X)
The ocular lens (eyepiece) is 10X
Total magnification= objective x ocular
 Magnification
Magnify/Magnificaton: to enlarge something in
appearance.
3 Objectives:
1. Scanning (4X)
2. Low (10X)
3. High (40X)
The ocular lens (eyepiece) is 10X
Total magnification= objective x ocular
 Staining
Stains are used in microscopy to increase contrast or color in
specific components of a specimen

Stained Red Blood Cells Gram Positive Bacteria:

streptococcus pneumoniae
Clonorchis sinensis
Mosquito
Lice
 Handling of Microscope
• Use lens paper ONLY to clean microscope
lenses
• Start ALL observations on the lowest
powered objective (4X)
• Only use coarse focus on the low power
(10X) objective and fine focus on high
power (40X) objective
Lab Materials and Trouble Shooting

Microscope Slides Cover Slips

 Today’s Lab
• PART A:
– Table 1, pg.73
– “e” slide (pgs. 74-75);
– # 12, (Desmids OR Rizophus)
– Preparing Wet Mount : 1 drop of Chlorella algae (pg. 76)
– Using a Biological Stain (78): WARNING: Calberla will
stain!!!; gloves can be worn if needed; 2 drops of pollen on
slide, but only stain 1; during the 5 min it takes to stain you
can start on Part B
– Table 2, pg. 79
 Today’s Lab
• PART B: The Dissecting Microscope
– Figure 1
– “e” slide
– Pg. 71; Total of 4 specimens: Duckweed, Peacock
feather, Beetle, or Printed Photo (view and draw
only 1 specimen)
 Today’s Lab
• Part C: The Electron Microscope (81)
– Examine the photomicrographs in the packet, to
fill in specimens for Table 3.
– For questions below Table 3, use
photomicrograph (Cat Flea)

– All of this must be checked off before you leave

today.
 Today’s Lab
• PART D: Clean Up (pg. 74)
– Wash hands
– TURN OFF ALL SCOPES AND BE SURE THEY ARE SET TO
THE SCANNING (LOWEST) OBJECTIVE
– Clean all microscope stages
– Rinse all slides with DI water
– Used cover slips go into the Broken Glass Container (NOT
IN THE TRASH)
– Transfer pipets can be trashed
– Clean cubbies and throw away any other trash
 HOMEWORK
• Work on Allelopathy Lab Reports
• Read Lichens and the Environment Lab.
• Remember Next Week IS AN OUTSIDE LAB