Laboratory Practice 3.

Observation and comparison of cells under a light microscope.

Introduction to Experimental Science Clarisa Blancas #579387

Andrea Díaz Conty Colunga Andrea Rodriguez De La Cerda #57944
Room 1202
Second semester
 Name: Clarisa Blancas , Andrea Rodriguez De la Cerda ID:584396
Date: 13/03/2019 Room:1202
Laboratory Practice 3. Demonstration/Quantitative. ​Observation and comparison of cells under a
light microscope.
Purpose:
To use a light microscope to observe different types of cells. Register obtained data and draw cell
structures.

Introduction:
According to Robinson (2018), plant cells are classified as eukaryotic cells, which contain a
nucleus. Plant cells differ from other eukaryotic cells because they contain chlorophyll and
organelles are the major parts of the cell. Plant cells are different from animal cells. Plant cells
have rigid cell walls, unlike animal cells that have more flexible cell membranes. ​The main
structural differences between plant and animal cells lie in a few additional structures found in
plant cells. These structures are: chloroplasts, the cell wall, and vacuoles.

Our investigation question is “What is the type of onion with the largest cells?

The importance of this is investigation is to understand the importance of cell length and differ
between the cells´ lengths of spring and white onion. Onions have an epidermal cell, which has
a ​simple structure, is very transparent, and its cells are easily visible under a microscope. That
is great to use in order to understand ​plant anatomy​, which is the study of the internal​ ​structure
of ​plants​. ​This experiment helps to learn how to differienciate between animal and plant cells,
since plants cells are normally larger. According to Bailey (2018) ​the normal length for an
animal cell varies from 10 to 30 micrometers, while for a plant cell is from 10 to 100
micrometers.

Theoretical framework​:
According to Daniel J, Cosgrove (2005), plants cells have complex walls,they are eukaryotic
cells, meaning that they have nucleus with specialized structures called organelles that carry
out different functions. Animals, fungi, between others also have eukaryotic cells, meanwhile
bacteria and archaea have simpler prokaryotic cells.

According to Biology Dictionary Editors (​2017), ​plant cells are differentiated from the cells of
other organisms by their cell walls, chloroplasts, and central vacuole.The plant cell has many
different parts. Each part of the cell has a specialized function. As previously mentioned plant
cells have a cell wall, a large vacuole, it has chloroplasts. None of these can be found in
animal cells, since animal cells do not have a cell wall or chloroplast and they either have a
small or no vacuole. We can identify some characteristics that are in both types of cells, like,
they both own mitochondrion, golgi apparatus, and a nucleus.

According to the Better Health Channel (​2015)​, spring onions have vivid green stems.The
vegetable also gives an onion flavour and crunch when raw. The Egyptians, Greeks, Romans
and Chinese all planted and used this vegetable. Spring onion appeared in Chinese history
since 2000 BC. This vegetable is known with powers to cure colds, and improve poor eyesight.
In the other hand we have white onions, white onions due to the compact nature of their cell
structure, they do not store quite as long as other varieties.

H​ypothesis:
Spring onion cells will be bigger in size that the white onion cells.

Dependent variable: ​What is measured, specifying units, measuring instruments, and uncertainty.

Size of cell measured with Logger Pro (+- micrometers).

Independent variable: ​What is manipulated or changed, the factor that is tested in the experiment.

Type of onion (Spring and white onion).

Controlled variables: ​The conditions that must remain constant and the ways to control them
(specifying units, measuring instruments, and uncertainty).

The same dye (methyl in blue) to stain the onion samples, the same amount of drops added to the
onion samples (1 drop), the same microscope was used to observe the onion samples, and same
app used to measure the onions´ cells (Logger Pro).

Materials and substances:

● Compound light microscope
● Glass slides
● Cover slips
● Forceps
● 1 white onion
● 1 spring onion
● 1 banana
● 1 slice of liver (cow or chicken)
● 1 carrot
● 1 grass leaf
● Pond water
● Scalpel
● Spatula
● Toothpicks
● Methylene blue dye
● Iodine dye
● Distilled water
● Digital camera or mobile phone camera
● Computer with Logger Pro software from Vernier App. 3.10
Safety precautions:

Remember to always follow laboratory rules and policies before beginning the practice.
Have special care while manipulating the dyes to avoid staining your hands. The professor
will provide precise instructions.

Methodology:
The method used to observe cellular structures using a compound light microscope is generally
the same for the different types of tissues.
1. Place the cells on the microscope slides in a layer that is no more than one cell thick.
2. Add a drop of water or dye as required.
3. Carefully place a cover slip over the drop. Try to avoid trapping air bubbles between the slide
and the cover slip.
4. Remove excess liquid or dye by placing the microscope slide inside a folded paper towel and
apply a light pressure on it.
5. Begin to focus observing the sample using the smallest objective lens, the one with the lowest
power.
6. Move the stage to obtain the most promising areas in the middle of the sample and then use
higher power objective lenses.
7. Draw and label the observed cells. (Mindorff, D. 2007)
8. The tissues to observe are the following:
NOTE: All the lab teams will work with both types of onions, besides, the lab teacher will assign another
tissue per table.
Tissues Required dye Observations

White onion Methylene blue Separate one of the internal layers of the onion and detach
the soft thin membrane that is adhered to the concave
interior layer. Take a picture of the focused image using the
10X objective lens, this way this image will be magnified
100X its actual size.

Spring onion Methylene blue Separate one of the internal layers of the onion and detach
the soft membrane that is adhered to the concave interior
layer. Take a picture of the focused image using the
objective lens 10X, that is, this image will be magnified 100X
its actual size.

Banana Iodine/Lugol Peel the banana and with a toothpick scrape the internal
pulp, without seeds. Smear the pulp on the toothpick onto a
microscope slide.

Carrot Methylene blue With a scalpel, make a very thin cut and place it on the
microscope slide.

Liver Methylene blue With a toothpick, scrape the liver. Smear the sample on the
toothpick onto a microscope slide.

Cheek Methylene blue Each student will gently scrape the inside lining of one cheek
 with a toothpick. Smear the sample on the toothpick onto a
microscope slide.

Grass Drop of water Place a small leaf directly on a microscope slide.

Pond water No dye Place one drop of pond water directly on a microscope slide.

Preparated slide Immersion oil Place the prepared slide directly on the microscope stage.

Draw, color, and label your observations of the structures the cells of each tissue:
Second Session: ​To be​ ​completed by the student.

In this session the student will obtain, register, process data and write a lab report that includes a
conclusion and an evaluation of the methodology used to estimate cell size of a specific plant tissue. Also,
compare the cell size of the two different tissues.

1. Process the white and the spring onion photographs obtained in the first session.
2. Image processing:
● Open the onions photographs and save the images in the computer as a .jpg files, without
changing the magnification or zoom. Do not increase or decrease the image size.
● Verify that the selected field contains at least 10 complete cells in it.
● Open the saved picture with Paint software. Use a metric ruler and the mouse to draw a 1 cm
line right below image. Open a text box below the line and write the length the scale line
represents: 100 micra. Save as a .jpeg file.
● Open the Logger Pro software Version 3.9 that is set up in the lab computers. Retrieve each
photograph of the onions with: ​Insertar / Imagen / con foto análisis.
● To establish the scale, click on the ​yellow horizontal ruler and drag the cursor with the
mouse along the scale line and let it go. Write the size that the scale represents in the text
box: 100 micra and click on ​Aceptar.
● To measure each cell, click on the yellow ​vertical ruler (with the red arrow) and drag the
cursor with the mouse from one end of a cell to the other, and let it go.
● Make the measurements of the length of the 10 selected cells.
● The labels with the lengths of the cells can be dragged out of the field where the cells are.
● Now, ​Print/Page the screen with the image of the 10 measured cells and save in a Word
document.
● Record the data in Table 1 of the lab report. Attach the print/page images to the lab report.
● Construct a graph which shows the mean values of the cells lengths or widths and includes all
the requirements.

Chemical disposal​:
When completing the lab practice, discard the samples as indicated by the professor.

Raw data registration:​.

Table1. The length of the spring and white onion´s cells. (micrometers)

Sample White onion Spring onion

Length in µm (± 0.1 µm) Length in µm (± 0.1 µm)

1 145.2 163.9

2 121.9 141.9

3 125.3 188.8

4 170.7 192.5

5 166.1 188.8
 6 196.9 136.9

7 130.1 107.9

8 138.7 140.8

9 128.1 81.7

10 149.5 166.6
AVERAGE ​ ​ 147.2 150.9

comment: ​We can observe from the table above that the length of the cells in the spring onion column
and as well as the white onion column have no pattern nor they follow a rule. The data that is recorded
shows different values and at the end of the table we can observe that the final averages, spring onion
obtained a higher value than white onion by a difference of 3.7 micrometers

Result processing and analysis:

Graph 1: The average length of the spring and white onion´s cells.

comment:
The averages observed in the graph above are clearly identified by the height of the bars, that
the two types of onions can be differed in the color that they are shown. The color blue is
identifying the white onion and the color green identifies the spring onion. We can also observe
that the average length is greater for the spring onion than the white onion. There is a value
difference between them of 3.7.

Conclusions:
The hypothesis was accepted, since as stated: spring onion cells will be bigger in size that the
white onion cells. In the majority of the 10 cells that were measured, the spring onion cells´
lenght was larger than the white onion´s one. The mean length of the spring onion cells´ was
150.9 micrometers (+- 1 micrometer) and for the white onion´s was 147.2 micrometers (+- 1
micrometer). By doing this experiement, we can learn how plant cells look like by identyfying
their cell wall under the microscope and also by observing the lenght of the cells are inside the
plants´ cell range as stated in the theorical framework. This helps us to know how to
differienciate animal and plant cells according to their size. It was proven correct that the thin
layer of the onion (epidermal cell) is very easy to use in order to understand a plant´s cell
structure because of its transparancy and large cells.

References​:
Allot, A & Mindorff, D. (2015). ​Biología. Programa del diploma​. Reino Unido: Oxford University Press.

Cosgrove, D. J. (2005, November 01). Growth of the plant cell wall. Retrieved from
https://www.nature.com/articles/nrm1746

Department of Health & Human Services. (2015, October 17). Spring onions. Retrieved from
https://www.betterhealth.vic.gov.au/health/ingredientsprofiles/Spring-onions

Seasons, Colors, Flavors, & Sizes of Onion. (n.d.). Retrieved from

https://www.onions-usa.org/all-about-onions/colors-flavor-availability-and-sizes-of-onions

Biology Dictionary ​Editors. (2017, April 29). Plant Cell - Definition, Functions and Structure.
Retrieved from ​https://biologydictionary.net/plant-cell/

Robinson, P. (2019, March 02). The Cell Structure of an Onion. Retrieved from
https://sciencing.com/cell-structure-onion-5438440.html

Bailey, R. (2018, October 10). Differences Between Plant and Animal Cells. Retrieved from
https://www.thoughtco.com/animal-cells-vs-plant-cells-373375