1. Laboratory Introduction

2. My Rules for the Laboratory
Be ready for the laboratory exercise you are performing that day! Read the exercise introduction.
Be on time for the laboratory! I will start the laboratory ON TIME with a lecture or a discussion covering what we are doing that day (a foundation).
I will check, periodically, to see if you are keeping up with doing lab exercises and reports.

3. Figure 3.1 Why does the abundance of water allow life to exist on the planet Earth?

4. Aseptic Technique
Whenever working in the lab, special care should be taken…
Use a soft hand; a gentle touch
Everything you use should be sterile and clean
Solutions should always be prepared fresh
Transfer-ware should never be reused
Containing-ware should be clean
Cadaver specimens are handled with gloves

6. Now lets look around the lab and identify our reagents and equipment…
Microscopes and Slides
Skeletal and Muscular models
Neurological Models
Tissue specimens

7. “Care and Use of the Microscope”
Laboratory Exercise
“Care and Use of the Microscope”

8. Care and Use of the Microscope
In this exercise, we will study the application of a compound and a dissection microscope, know their parts and function, and learn to use and take care of them.
These terms should be familiar to you: magnification, resolution, stereoscopic- (dissection), simple-, compound-, and electron- microscopy (scanning or transmission).

9. What procedures are we doing?
We will know the parts of-, the proper use of- and the clean up procedure for a bright-field compound and dissection microscope.
You will examine a prepared slide and a slide with the letter “e” affixed to it using a compound microscope.
You will examine a sample of your own hair using a dissection microscope.

10. Resources available…
Anatomy and Physiology Laboratory Study Pages; submenu: Microscope

11. Laboratory Lecture

12. Concept: Cell Biology tools - microscopy & chemistry
The quality of an image depends on
Magnification, the ratio of an object’s image size to its real size
Resolution, the measure of the clarity of the image, or the minimum distance of two distinguishable points
Contrast, visible differences in parts of the sample (can be enhanced by stains/markers)

13. -Electron Microscope 2 nm
Scale of Resolution
-Naked Eye .2 mm
-Light Microscope .2 mm
-Electron Microscope 2 nm
10 m
Human height
1 m
Length of some nerve and muscle cells
0.1 m
Unaided eye
Chicken egg
1 cm
Frog egg
1 mm
100 µm
Most plant and animal cells
Light microscope
10 µm
Nucleus
Most bacteria
1 µm
Mitochondrion
Figure 6.2 The size range of cells
Smallest bacteria
Electron microscope
100 nm
Viruses
Ribosomes
10 nm
Proteins
Lipids
1 nm
Small molecules
0.1 nm
Atoms

14. Light Microscopy
In a light microscope (LM), visible light passes through a specimen and then through glass lenses, which magnify the image
Various techniques enhance contrast and enable cell components to be stained or labeled
Most subcellular structures, including organelles (membrane-enclosed compartments), are too small to be resolved by a Light Microscope

15. Viewing Techniques for: Imaging
Naked Eye
Light Microscope
Electron Microscope
TECHNIQUE
RESULTS
(a) Standard/Brightfield
(unstained specimen)
Imaging
50 µm
(b) Standard/Brightfield
(stained specimen)
Imaging w/ stain
(c) Phase-contrast
Imaging & density
The microscope manipulates optics to improve contrast between the structures
(d) Differential-interference-
contrast (Nomarski)
Imaging & optics
(e) Fluorescence
Imaging w/ Labeling
Figure 6.3a-d Light microscopy
50 µm
(f) Confocal
Imaging & focal planes
50 µm

16. Electron microscopy
Two basic types of electron microscopes (EMs) are used to study subcellular structures
Scanning electron microscopes (SEMs) focus a beam of electrons onto the surface of a heavy metal stained specimen, providing images that look 3-D
Transmission electron microscopes (TEMs) focus a beam of electrons through a heavy metal stained specimen, used mainly to study the internal structure of cells

17. (b) Transmission electron
TECHNIQUE
RESULTS
Cilia
1 µm
(a) Scanning electron
microscopy (SEM)
SURFACE
Longitudinal
section of
cilium
Cross section
of cilium
1 µm
Figure 6.4 Electron microscopy
(b) Transmission electron
microscopy (TEM)
SECTION OR SLICE

18. A quick note…
I have presented an introductory review of laboratory Exercise 1. This is because the class has just started and I want you to get off on the “good foot.” From now on, you are responsible for reading the introduction before the exercise. All the stuff we do in lab is covered in theory during a lecture (BIOL2301) and is also covered on the internet on the Anatomy and Physiology Lab Pages (let’s take a look at awesome central!):

19. Lets get familiar with each other; we will be working in groups to complete exercises (3 groups total in this class – groups A, B and C).
Finally, lets look in the laboratory manual... Within your group, who is the group leader for the Exercise? Who is going to do which experiment(s) of that Exercise? You must decide amongst yourselves. Next step: decide who is going to get the materials to complete the exercise. This is collaboration!