Cellular biology

Practical Problem 1: How to observe cells under a microscope

1. Objectives

- To know how to use a properly use a microscope

- To distinguish several cell types under microscope.

2. Introduction

All living organisms are made up of cells.

Cells are the smallest part of a living organism and are around 0.01 mm - 0.03 mm long.

To look at a cell close up we need a microscope.

Microscope, instrument that produces enlarged images of small objects, allowing the observer an
exceedingly close view of minute
structures at a scale convenient for
examination and analysis. Although
optical microscopes are the subject of
this practical, an image may also be
enlarged by many other wave forms,
including acoustic, X-ray, or electron
beam, and be received by direct or
digital imaging or by a combination of
these methods. The microscope may
provide a dynamic image (as with
conventional optical instruments) or
one that is static (as with conventional
scanning electron microscopes). The
most familiar type of microscope is the optical, or light, microscope, in which glass lenses are used
to form the image. Optical microscopes can be simple, consisting of a single lens, or compound,
consisting of several optical components in line. The hand magnifying glass can magnify about 3
to 20×. Single-lensed simple microscopes can magnify up to 300×—and are capable of revealing
bacteria—while compound microscopes can magnify up to 2,000×. A simple microscope can
resolve below 1 micrometer (μm; one millionth of a meter); a compound microscope can resolve
down to about 0.2 μm.

3. Procedure
3.1 How to use a microscope
Move the stage (the flat ledge the slide sits on) down to its lowest position.
Place the glass slide onto the stage. Be careful pushing it under the clips that the cover
slide doesn't move or crack.

Select the lowest power objective lens.

Turn the coarse focus knob slowly until you are able to see the cells.

Turn the fine focus knob slowly until the cells are in focus and you can see them clearly.

Repeat steps 1-5 using the higher power magnification to see the cells in more detail.

3.2 Calculating magnification

To see an object, the eye piece lens and the objective lens magnification are multiplied
together to give the total magnification.

Total magnification = eye piece lens magnification × objective lens magnification

For example: 10 × 20 = 200

Magnification = 200

3.3 Observe cells under a microscope

Try to observe several cell types under the microscope
4. Report
Describe which cells types were observed under the microscope:
Total magnification:
Determine the kingdom to which the cells belong:

Make a comparison of these cells.

 Cellular biology
Practical Problem 2: Observing cell division and tonicity of the
cell
1. Objectives

- To observe cell division stages from onion root tips

- To better understand the tonicity of the cell

2. Protocol
2.1 Protocol for chromosome spreads

Step 1: Collect the onion root tips

Step 2: Pretreatment the root tips with HCl 2M in 10 minutes, at 40oC. Then wash by water 2
times.
Step 3: Staining the root tips with 1 drop of 1% Toludin blue for at least 2 minutes
Step 4: Remove the excess stain, wash 2 times by water
Step 5: Cut the root tips in to 3mm piece
Step 6: Gently transfer 1 piece of root tip in 10 µl of 45% acid acetic laying on the slide. Put the
cover slip and observe under microscope with 40x or/and 100x objectives with oil immersion.

2.2 Test the tonicity of the cell

Tonicity describes the capacity of a cell to lose or gain water. It depends on the quantity of solute
that does not penetrate the membrane. Tonicity delimits the magnitude and direction of osmosis
and results in three possible scenarios that alter the volume of a cell: hypertonicity, hypotonicity,
and isotonicity. Due to differences in structure and physiology, tonicity of plant cells is different
from that of animal cells in some scenarios.

To test the tonicity of the plant cell, we can perform the following procedure

Step 1: Prepare 10ml Sodium chloride solution at difference concentrations: NaCl 3M, NaCl 0.3M,
NaCl 0.03 M, and water

Step 2: Transfer 1 thin layer of onion on the slide

Step 3: Cover the onion samples by 1 drop of sodium solution. Wait for 5 minutes

Step 4: Put the cover slip and observe under the microscope.
3. Report
Describe what were observed under the microscope:

- Base on your observation, which cells are in the dividing stage?

- Describe the shape of cells in different solutions. What is your idea about the
effectiveness of ion concentrations on cells?