1. Microscopical analysis
BIOL30002 Experimental Reproductive Physiology
BIOL30002 Experimental Reproductive Physiology
Techniques: Microscopy
Microscopical analysis
Structure   Function
Gross anatomy (macroscopic) tells us what is happening on a gross scale
Microscopical anatomy tells us what is happening on a cellular scale

2. Microscopy Scale limited by detection method
visible light: wavelength ~ 0.38 – 0.74 μm
 cannot reveal structures less than about 1 μm
(organelles)
Electron beam: at 10kV  μm
(large molecules)
Synchrotron beam:  μm
(small molecules)
Atomic force microscopy  atomic scale

3. Types of microscopy Light microscopy
thin sections – paraffin; frozen – 4-10 μm
good for tissue/cellular level
relatively low cost
many ways to stain to highlight things of interest
Confocal microscopy
light microscopy with special optics
can get higher resolution
can build up 3-D structure
Transmission Electron microscopy
extremely thin sections (0.05 μm)
good for cellular and subcellular level
expensive machinery
more limited staining options (heavy metals)
Scanning EM
shows surface structures

4. Interpreting Microscope Slides
2-D vs

5. histology light microscopy vs TEM vs SEM vs confocal stains Resources:
haematoxylin & eosin - good general purpose
trichrome - good connective tissue discrimination
PAS - sugar containing components
lipophilic dyes - fats and oils
nucleic acid stains
enzyme histochemistry
immunohistochemistry
Resources:
Kerr (2000) Atlas of Functional Histology
Wheater et al, (1987) Functional Histology

6. Basic tissue types blood connective tissue, bone fat (adipose tissue)
muscle
nerves
epithelium (epithelia)

7. epithelia columnar cuboidal squamous … lumen
Junctions between cells  barrier
columnar
cuboidal
squamous …
lumen
uterine luminal epithelium

8. pituitary
stalk (with blood vessels)
posterior
(nerve ends)
anterior

9. testis
epididymis
testis
fat
epididymis

10. BIOL30002 Experimental Reproductive Physiology
Techniques: Microscopy
seminiferous tubules
lumen
blood
germ cells

11. BIOL30002 Experimental Reproductive Physiology
ovary
Techniques: Microscopy
antral follicle
oocyte

12. Drawings Use a SHARP pencil
Tissue outlines – don’t attempt elaborate shading unless you are very skilled
Use appropriate size scales
Don’t spend forever replicating fine detail – just do a bit to show
Label and annotate with observations and info from reference material