General rules in biological experiments

u  No eating and drinking in the lab

u  Always keep your bags and other belongings in/on
the locker on the window side. Never place it on the
floor because there is a danger of chemical
contamination.
u  Excessive makeup should be refrained. Do not refrain
from things that may scratch the microscope lens,
especially mascara.
u  Wear clothes easy to move. No high-heeled shoes,
sandals and mules, and highly exposed clothing.
u  No use or charging your mobile phones in the lab. It
may be confiscated.
How to use microscope
 Before starting, check
body tube�

1)  is the body tube stable?

2)  is the objective lens

screwed in tightly?

3)  any abnormalities?

Objective lens�
Stage�

Do NOT touch the lens！

 Connecting the electric cable

Power switch

OFF

dimmer control dial

Before connecting the electric cable, make sure

1.  the power switch is in OFF position(⭕),
2.  and the dimmer control dial is fully turned toward the
front
 Connecting the electric cable

insert the power cord

to the AC-IN socket on
the back of the
microscope and insert
the plug to a power
outlet on the lab bench.

AC-IN socket
Microscope operation procedures

•  Follow the instruction on IV-9 of

Biological Science in the textbook.
•  Samples are on the bench.
•  Pay particular attention to IV-9 (7)-(11)

Do not let the objective lens collide

with the sample!
 Get a focus with low mag. lens (10x)
place the 10x objective lens in the light path by rotating the revolver.
Lower the stage with the focus knob

Revolver

Do NOT rotate with

holding a lens!

Focus knob
(fine adjustment)
Focus knob
(coarse adjustment)
 Setting a sample slide�
place a slide firmly on the sample holder�
①� ③�

②� ④�

①② move the sample clip

③ place a slide glass
④ move the clip back to the original position,
then confirm the slide is stably fixed
 Adjusting the position of the slide�
Move the stage with the stage movement knob
to place your sample is right above the condenser lens

Stage movement knob�

sample�
Rotate the focus handle slowly to raise the stage while looking at the
microscope from the side, so that the spacing between the objective
lens and cover glass gets to within 5mm, being careful not to hit the
cover glass with the objective lens.

Focus knob
(coarse adjustment)

looking from the side�

Focus knob
(fine adjustment)
Turn on the power switch.
Slowly rotate the dial clockwise and while looking through
the ocular lens to adjust to the appropriate brightness.

Power switch
　 ：ON
○：OFF�

Power switch

Dimmer control dial

Look through the ocular lens, and move the observation
tube to adjust the distance between ocular lenses to
your eyes.

Two fields are off-centre You see only one field

Look through the ocular lens and rotate the focus knob
slowly to lower the stage until focused on the sample
Once something appears in the field of vision, turn the stage movement
knob a little, and if what is seen moves then the microscope is almost
focused on the sample surface

Focus knob
(coarse adjustment)
Stage movement knob�

Focus knob
(fine adjustment)

Do not rotate the focus handle in the wrong direction!

the cover glass and objective lens will strike against each other.
 Next, place the 40x lens in the light path.

Rotate the revolver to place the 40x lens in the light path.

DO NOT move the stage!

If it is focused properly with 10x lens, 40x objective
lens will not hit the sample slide. Do not move the
focus knob when rotating the revolver.

After setting 40x lens, move the stage to place

the specimen you wish to observe in the centre.
- You will see the central area of the field is now magnified.
- If not focused properly, move the focus knob (fine
adjustment) slowly until the focus is perfect.
- If you find the field too dark, adjust the brightness with
the iris diaphragm

Focus knob
(fine adjustment) Iris diaphragm
Observing animal tissue
 Objective of this class
•  Acquire good understanding of basic handling of a
microscope and how to perform microscopy.

•  Observe tissues of flog back skin. You are expected to

learn how a collection of cells form a tissue.
•  Sketch an epithelial tissue and a connecting tissue that
form skin. Epithelial tissue is Pay attention to how the
two tissues are organised.
Epithelial tissue: The cells form continuous sheets which are attached to
each other at many locations.
Connective tissue: Connective tissue is a group of unstructured cells and
connects between tissues.
Skin tissue of back side of flog (low mag.)
Epidermis Mucous gland�
(epithelium)�
Pigment cell� Efferent duct�
Skin1
(surface)�

Granular gland�

Stratum
spongiosum�

Stratum Spongey layer�

compactum�
Stratum spongiosum�
Skin2
(inside)� Stratum compactum�
 Skin surface1
Skin is made primarily of
epithelial tissue and Epidermis
connective tissue and (epithelium)�
covers the body surface.
The outside layer M
consisting of epithelial
海綿層
tissues is called
Epithelium. The inner
layer called Stratum
Stratum spongiosum
spongiosum consists of (sponge layer)�
connective tissues and
develop secretory glands. G
 Skin surface 2
Epidermis has multi-layered Horny layer�

structure (stratified squamous Migratory layer�

epithelium). Germinal layer�

The lowest layer is the germinal

layer (GL), where cells
Epithelium�
proliferate (mitotic cell division).
Proliferated cells move to the
migratory layer and then form
the horny layer where old cells
come off as dirt.
With this mechanism, skin can protects human
body from pathogens etc.
 Deep inside the skin（connective tissues）
Granular gland�
Spongy layer�

Compact layer�

Compact layer�

Below the sponge layer deep inside the skin is a bellows-like

connective tissues called a compact layer.
 Report assignment
(Basic 1)
Measuring lengths under microscope with a
micrometer and sketching animal tissues

How to use micrometer
Adjusting angle of ocular micrometer
•  Ocular micrometer is already set in one of ocular lens (usually
in the right)

•  Rotate the top ring part (yellow arrow) to adjust angle of ocular
micrometer.

10

20

30

40
0 10 20 30 40 50 60 70 80 90 100

50

60

70

80

90

10
0
Length measurement using ocular micrometer
In actual measurement,
(1)  rotate the ocular micrometer.
(2)  move the position of the sample using the stage moving knob
together to place the scale and the sample in parallel.
(3) align one end of sample to a scale line of ocular micrometer.
(4) read the scale to count the number of scales for the sample.
(5) then, convert to length.
* When reading the scale, read up to 1/10 of the minimum scale.

0 0
10 10

90 90

80 80

70 70

60 60
0 10 20 30 40 50 60 70 80 90 100
50 50

40 40

30 30

20 20

10 10

0 0

Rotate ocular micrometer� Move the sample�

Assignment 3（Measure the thickness of epithelium）
Take measurement of thickness of epidermis (one point) in the skin
using a micrometer. Use 40x objective lens and set one minimum
scale to 2.5µm. Convert your reading to the actual thickness (○○. ○
µm) and write it at the top of your sketch (on one of sketch papers at
the end of the text).

HL
ML
GL

All assignments (1-7) are uploaded to the Moodle system.

Do all assignments after the class.
 Assignment 4 & 5（sketch）
[Assignment 5] observe a slide of skin
tissue and draw a sketch.

Sketch should include all tissues and

structures described under the skin
海綿層
images in page IV-31, 32, except for
⑥efferent duct and ⑧blood vessel.
顆粒腺

上皮
2. switch to 40x objective (high
mag.) and add all the details of
tissues and structures.
海綿層

[Assignment 6] insert names of each

tissues and structures. Use leader lines
as in the example where appropriate.

Your sketch should include at least one detailed drawing for all structures and tissues.
 How to sketch (Assignment 4)

First, decide an area you want to sketch.

Choose an area that incudes at least

one each of all tissues and structures Low mag. image (with 10x objective lens)
in IV-31-32.

The thickened region is often a good

area for this assignment.
 Assignment 5（sketch）
1. draw outline of the sketching area
with the 10x objective (low-mag.) and
set the position and size of each
tissue. Double-check your sketch
meets all requirements of the
assignment. 海綿層

顆粒腺

上皮
2. switch to 40x objective (high
mag.) and add all the details of
tissues and structures.
海綿層

Your sketch should include at least one detailed drawing for all structures and tissues.
 Assignment 6（insert a scale bar）
Suppose the thickness of the epithelium is 27 µm.

海綿層

1.  Measure the thickness of the 顆粒腺

epithelium in your sketch in
mm. 上皮

2.  Calculate the length

equivalent to 100µm in your 海綿層
sketch and insert a scale bar
of 100µm under your
sketch.
 Time table (Basic 1)

Microscopy and sketching

Setting up a Introduction Tidy up and

miscroscope of today’s leave
assignments HL
•  ML and sketching.
You can leave once you finish microscopy
But remember to take the sketch paper home and finish all other
assignments. GL
•  Start tiding up after 80 min even if you have not finished the
assignments and leave the room at the end of the period
(90min). You may capture microscope images of the specimen
with your phone etc. to bring back to your home.
 At the end of today’s experiment

•  Remove the specimen slide from the
microscope.
•  Set the dimmer control dial to the lowest and
switch off the main power.
•  Wash your hands at the side sink.
•  Collect all your belongings (pens, report papers
etc.) and leave the room quietly.
•  Leave the microscope and other equipment on
the bench.
•  Avoid unnecessary touching of equipment.
Staffs will disinfect them.
 Diopter adjustment
1. Set the diopter adjustment ring in the central position
2. looking through the right objective lens with your right eye
(keep the left eye closed)
3. Rotate the focus knob to focus on the sample
Pay attention to the rotating direction!

Focus knob
(coarse adjustment)
Stage movement knob�

Focus knob
(fine adjustment)
 Supplement

•  Refer the next slide only when you need.