EXPERIMENT 3.

1
PREPARING A SLIDE OF PLANT TISSUES
OBJECTIVES
1. To make a thin slice by using hand-cutting method
2. To prepare, stain and make a aslide of plant sapmle
3. To illustrate the observation of plant sample

MATERIALS
Samples
A young dicot
Apparatus
Compound microscope
Slide and coverslip
Chemicals
Eosin

METHOD
1. About 2 cm of a young root of balsam is obtained. It is placed on microscope
slide or plate
2. 2. The root is cut very thin through the region of maturation
3. The thinnest section is selected and it is transferred to a slide that contain a
drop of water.
4. The slide is observed by using compound microscope. The observation is
sketched and labelled in the space provided in Table 3.1
5. The tissue system in young root system; vascular cylinder is identified.
6. Step 1 until 4 is repeated by using stem of dicot.








EXPERIMENT 3.2
PREPARING A SLIDE OF INSECT TISSUES
OBJECTIVES
1. To make a slide by using squash technique
2. To prepare,stain and make a slide on insect sample
3. To illustrate the observation of insect sample.
MATERIALS
Sample
A male locust (Example: locust, Valanga nigricornis)

Apparatus
Dissecting and compound microscope
Slide and cover slip
Small dissecting dish
Pin
Glass rod
Hotplate or Bunsen burner
Filter paper

Chemicals
Acetic-orcein
Canada balsam

METHODS
1. A freshly killed male locust was pinned out on a small dissecting dish with
dorsal side uppermost. The wings were pinned out on each side.
2. The abdomen was opened up by a mid-dorsal longitudinal cut. The body wall
was pinned back.
3. The testis was identified by using a dissecting microscope. Together with fat,
these made up an oval body lied above the gut in abdominal segment
(segments 5 and 6). The testes were transferred on a microscope slide.
4. The fat (yellow) was removed as much as can, only the white tubules of the
testes were leaved. Two or three were sufficient.
5. Several dropped of acetic-orcein stain was added and was put on cover slip.
6. Then, was covered with filter paper and was squashed gently. This helped the
chromosomes spread.
7. The slide was warmed up on a hot plate or Bunsen burner for about 10
seconds for the staining intensified.
8. The slide was observed using microscope, and specimen was sketched. A
photo was taken using digital microscope. The stages of meiosis were
examined. Chromosomes were drew and labeled in Table 3.2.
The slide was washed with a jet of water for the excess dye removed, then air
dried the material once again and was mounted in Canada balsam or similar
media.
















DISCUSSION

In Experiment 3.1, we have to make a thin slice of plant tissue sample
(Canada balsam) by using hand-cutting method and we also have to prepare
it as a slide that to be observed under light microscope. During the
experiment, we took two part of the plant, which are soft stem and root. We
chose soft stem as the sample because it makes the cutting method to be
easier. As a result, we can observe the structure of xylem and phloem in both
part of the tissue.

The characteristics of water-carrying cells(xylem) are it is in the middle
of the cell. The xylem of most vascular plants included tracheid, tube-shaped
cells that carry water and minerals up from roots. Furthermore, food-carrying
cells(phloem) placed at the outer layer of the cell. The phloem has cells
arranged into tubes that distribute sugars, amino acids, and other organic
product. Roots are organs that absorb water and nutrients from the soil.
Roots also anchor vascular plants, hence allowing the shoot system to grow
taller.

In Experiment 3.2, we have to make a slide of incests testis
(grasshopper) by using squash technique and preparing a slide for observing
under light microscope. First, we have to choose a male grasshopper to be
examined. Then we have to pin it on the blue wooden board. Next, by using
dissecting knife, we cut of longitudinally its abdomen and the testis (needle-
like shape) is taken out by using forceps and it was placed on the glass slide.
Then we added several drops of acetin-orcein stain and put on the cover slip.
The observations were portrayed it in the Table 3.2.
The precautions steps that we should engage in the slide-making process are
we have to cut as thin as possible the plant tissue so that it will be easier to
observe under light microscope. Furthermore, for experiment 3.2, once should
cut the insects abdomen longitudinally using a knife. Besides that, during
preparing a slide, it has to be warmed up 10 s on Bunsen burner accurately.

CONSLUSION
Using hand-cutting method for making a thin slice of tissue sample is not the
best way as the thinness of the tissue probably cannot be obtain accurately.







INTRODUCTION

Examining thin section of specimens using light microscope will be the
major tool to study cells, tissues and organ morphology. Histology involves
the preparation of the tissues for examination using a microscope.

Light microscope utilizes specific arrangements of lenses to magnify an
image. An electric bulb is the light source used to illuminate the tissue section.
The image is further magnified by the ocular lens of the eyepiece. This lens
focus the image on the retina of the eye. The total magnification of the image
depends in the magnifying powers of the objectives lens and that of the ocular
lens.

Quality of the image depends not only on magnifying capability of the lens but
also on its resolution. The resolution means of the lens to show that two
distinctive objects are separated.