LAB REPORT

EXPERIMENT 2

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 CELL STAINING AND MEASURING CELL SIZE
Safa Iwani binti Anuar Razif, Aqila binti Azahar, Nur Aina Syazwin binti Ramli, Nur Inas
Insyirah binti Madzlan & Muhammad Daniel Hakim bin Rosli (PI080S38)

INTRODUCTION

Cells are basic blocks built of all the living things. Human body is composed of many
trillions of cells. Cells have a lot of parts, which are every each if them have their own
different function. There are some cells contain a number of functional structures called
organelles which found in eukaryotic cells. Organelles carry out functions such as making
protein, processing chemicals and generating energy for the cells. Shapes of cells are
depending on the particular functions that they have to do.

From this experiment, we observed that the structure of stained plant and animal cells
using microscope since their sizes are too small for our human eyes to see. We used plant
and animal cells because they consist of certain similar and different part within cells. The
both of the cells are differ in shape, so, we could compare the differences between these two
cells.

OBJECTIVES
1. To prepare a wet mount plant cells and animal cells.

2. To compare stained plant and animal cells under the microscope.

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MATERIALS AND METHOD

Materials
Allium sp.(onion), iodine stain, Methylene blue
Apparatus
Scalpel, graticule, forceps, stage micrometer, microscope slide, microscope, cover slip,
toothpick

Method
Dependant variables:
1. Image of plant cells
2. Image of animal cells
Independent variables:
1. Plant cells
2. Animal cells
Controlled variables:
1. type of animal cells
2. type of plant cells
Staining and observing plant cells
We cut a small section of an onion, Allium sp. Next, we peeled off the transparent and thin
epidermis of the onion by using forceps. We placed the onion epidermal in the center of a
microscope slide. We put one drop of iodine stain on the epidermis which produced a “wet
mount”. We placed one edge of a cover slip to one side of stain and slowly lowered the
cover slip. We also touched the edges of cover slip with paper towel to remove excess stain.
After that, we examine the onion cells under a light microscope.

Staining and observing animal cells

We added one drop of methylene blue in the center of a microscope slide. We used a
toothpick to gently scrape the inside of cheek. We smeared and mixed the cells in the
methylene blue solution. We placed one edge of a cover slip to one side of stain and slowly
lowered the cover slip. We also touched the edges of cover slip with paper towel to remove
excess stain. After that, we examine the cheek cells under a light microscope.

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DATA AND RESULTS

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DISCUSSION
For the onion cells, composed many cells which would have chloroplast present to the
process of photosynthesis. The general shape of an onion cell is square or rectangular. This
is because onion cell has a cell wall which maintain its shape. However, the onion cells do
not have chloroplast since onions are below ground and aren’t exposed to light too much.
Then, chloroplasts are not needed. When observed the onion cell, small dot can be seen.
There are no spaces between the onion skin cells because they were pushed together. The
Iodine stain are used in this experiment so the cell could be clearly seen under the
microscope and its part can be distinguished. The presence of starch in it make the color of
Iodine changes into blue dark. The number of cells that are seen in the field of view under
low power magnification were higher than the number of cells in the field of view under
higher magnification. The structure of cells looked clearer and able to study the cell when
observed the onion cells at the higher magnification. The nucleus, the cytoplasm and the cell
wall were the organelles that was observed under the microscope. The cheek cell, an
example of an animal cell, usually has an oval shape and also has no spaces between cells
because the cells were pushed together completely like the onion cells. Cells are
transparent. The methylene blue was drop in the cheek cell to make the cells appear clearly
and the nucleus pop up so the cells are easier to analysis. When observed the cheek cells at
the higher magnification, the cheek cells were more spread out from each other and they
had a round shape. The organelles that visible in this cell were the nucleus, the cytoplasm
and the cell membrane. However, mitochondria, lysosomes and endoplasmic reticulum are
present in cheek cells that are not visible in a light microscope. From the observation, we
can identify that both cells are Eukaryote because they have nucleus and organelles are
surrounded by a membrane. The different between cheek cells and onion cells are that the
chloroplast and cell wall only exist in the onion cells while the cheek cell does not .

CONCLUSION

From the result of the experiment, we can conclude that there is a difference between
stained plant and animal cells under the microscope. Cheek cells as one example of an
animal cell, do not show any sign of cell wall that surrounded the cell, as an animal cell
(cheek cells) only surrounded by semi-permeable cell membrane while for the plant cell
(onion cells), we can see that the cells are surrounded by the cell wall. Therefore, these
cause difference shape between the animal cell and animal cell due to the presence of the
cell wall in the plant cell. Cheek cells (animal cell) are in irregular shape while onion cells
(plant cell) are a rectangular shape. As for the similarities between plant cell (onion cells)
and animal cell (cheek cells), both are Eukaryotes as each of them has a nucleus. For the
measuring cell size, in the view under higher magnification, the structure of cells is more
clearer and sharper than lower magnification.

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REFERENCES

https://www.microscopemaster.com/onion-cells-microscope.html

https://schoolworkhelper.net/plant-animal-cells-staining-lab-answers/

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LAB REPORT

EXPERIMENT 3

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INVESTIGATING CELL MEMBRANE PERMEABILITY USING TUMERIC (Curcuma longa)

Safa Iwani binti Anuar Razif, Aqila binti Azahar, Nur Aina Syazwin binti Ramli, Nur Inas
Insyirah binti Madzlan & Muhammad Daniel Hakim bin Rosli (PI080S38)

INTRODUCTION
Small changes of the composition of cell membranes can strongly affect the functioning of
intrinsic membrane proteins, such as ion and water channels, which regulate the chemical
and physical balance in cells. Changes may occur due to the introduction of short-chain
alcohols, other anesthetics, at membrane surfaces.

Another aspect to the effect of alcohols appears in a more applied context. In the process
of producing alcoholic beverages, wine in particular, yeasts like Saccharomyces cerevisiae
have to sustain high ethanol concentrations without losing their viability. There is no
satisfactory understanding this effect. High alcohols concentration changes the membrane
structure and force transmembrane proteins into unfavorable conformation. In these
conformations, proteins cannot fulfill their functions and thus the yields drop dramatically.

For anesthetics, the situation is slightly better. Molecular dynamics simulations is used to
study molecular level mechanisms of general anesthesia using halothane as a specific
anesthetic. The concluded that the global effects of anesthetics due to generic interactions
mechanisms, are important and lead to modulations in the functions of channels and
proteins.

Phospholipid bilayers can be considered as a first approximation to understand the

behavior of cell membranes under the influence of alcohol, and much information can be
extracted from such systems. Our simulations show that ethanol is able to pass through the
bilayer much more easily than methanol. This can be explained by hydrophobic nature of the
carbon tail of ethanol, making passing through the hydrophobic chain regions of lipid bilayers
easier. Ethanol molecules condense near the interface region between lipids and the
surrounding water, there is sharply increased density of ethanol near the region of the
interface.

OBJECTIVES
1. To investigate the effect of ethanol on the structure of cell membrane of Curcuma
longa.
MATERIALS AND METHOD

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Materials
Turmeric (Curcuma longa), ethanol, distilled water
Apparatus
Scalpel, ruler, cork borer, 100 ml beakers, 10 ml measuring cylinders, tile

Method
Dependant variables:
1. Effect of ethanol on the structure of cell membrane of Curcuma longa.
Independent variables:
1. Concentration of ethanol
Controlled variables:
1. The type of cell membrane
2. The volume of solution in each beaker

For the experiment

We used a cork borer to cut six cylinders of fresh Curcuma longa tissue, and placed on a
tile. The cylinders were cut using a scalpel and a ruler to ensure they had the same length.
Then, the cylinders were soaked in water for five minutes. We made a 1:4 dilution of ethanol
and labelled three small beakers: (i) water (ii) 1:4 dilution ethanol (iii) 100% ethanol. We filled
each beaker with its respective solution, 10 ml each. Next, we removed the cylinders from
water and placed them onto paper towel to dry. Two Curcuma longa cylinders were added in
each beaker and we let them stand for 15 minutes. Lastly, we observe the colour of solution
in beakers.

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DATA AND RESULTS

Diagram 1

Solution Water 25% ethanol 100% ethanol

Observation There were no color The color of the 25% The color of the
changes in the ethanol changes to 100% ethanol
beakers the yellow changes to the
bright yellow

Table 1

Diagram 1 and Table 1 above show that after 15 minutes, distilled water in the first beaker
does not affect the permeability of turmeric ’s cell membrane. Water can pass through the
process osmosis easily. Meanwhile, in the second beaker the color of 25% ethanol changes
to the yellow. It was brighter than the first beaker. As for the third beaker, the color of 100%
ethanol solution changes to the bright yellow. It was the brightest compared to the others.

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DISCUSSION

In this experiment, we investigated the effect of ethanol on the structure of cell membrane
of Curcuma longa (Turmeric). We use different concentration of ethanol; water, 25% ethanol
and 95% ethanol on same length of turmeric. We know that the fluid mosaic model states
that a cell membrane is composed mainly of a phospholipid bilayer and embedded protein
molecules and these molecules are essential to maintain the structure and function of the
membrane, and the integrity of the contents within the cell. Plasma membrane only allow
selective materials to pass through it and this membrane can be denatured by specific
solvents, such as ethanol. Organelle that contains water is vacuole and it is surrounded by
membrane called tonoplast. In Curcuma longa, the vacuole contains a water-soluble yellow
pigment, curcumin, which gives the turmeric its color. Since, the pigment is water-soluble
and not lipid-soluble, so it remains in vacuole when the cells are in good conditions. If the
tonoplast is damaged, the contents of the vacuole will spill out into the surrounding
environment. In the case of the Curcuma longa, when the membrane is damaged, the bright
yellow pigment will leak out into the surrounding environment. The intensity of color in the
environment should be proportional to the amount of cellular damage sustained by the
turmeric.

In the first beaker, two Curcuma longa cylinders were placed into 10 ml of distilled water and
soaked it for 15 minutes. Water should not have any effect on the permeability of turmeric’s
plasma membrane and water also can easily pass through the plasma membrane by
osmosis and should not cause any damage to it, but after 15 minutes we observed that the
color of the distilled water turned slightly yellow. This happened because of the cell
membrane of Curcuma longa was already damaged when it was cut into using the cork
borer causing the yellow pigment to spill out and dissolved in the water

Next, in the second beaker, another two Curcuma longa cylinders were placed into 10 ml of
25% ethanol solution. After 15 minutes, the color of the solution turned yellow and brighter
than the first beaker. In this experiment, we found that ethanol has effect on the structural
properties of the membrane. Ethanol is a nonpolar solvent, and the phospholipid bilayer of
the turmeric plasma membrane is also nonpolar. Nonpolar solutes can dissolve in nonpolar
solvent and lipids dissolve in alcohol thus, the plasma membrane can be disrupted as it is
dissolved by the ethanol solution. Ethanol is able to form hydrogen bonds with the lipids in
lipid bilayer, and these hydrogen bonds reduce the order parameter of the lipid hydrocarbon
chains that means that the structure will be altered and there are more gaps in the
phospholipid bilayer, thus the membrane will be more permeable and more of the turmeric’s
yellow pigment will leak out into the ethanol solvent, causing the solution to become more
intense yellow color.

Lastly, when the Curcuma longa cylinders were soaked in 100% ethanol solution, the color
of the solution changes into bright yellow and the brightest compared to the others after 15
minutes. The color become brighter than the previous result because there are more
curcumin pigments that leaked out. The higher the concentration of ethanol, stronger the
effects of ethanol towards the plasma membrane. This is because higher concentration of

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ethanol damages the lipid and protein structure of phospholipid bilayer. Ethanol will dissolve
the lipids and also protein structure of plasma membrane and this will make the cell
membrane to become very fragile. This occurrence again causes the phospholipids
surrounding the protein to move about as there is more room and produce large gaps in the
layer to allow big molecules to pass. The cholesterol, as previously stated, was not soluble in
water but soluble in ethanol, such that in high concentrations it actually begins to dissolve.
As the cholesterol is no longer present and is used to hold the phospholipid together, the
bilayer become more fluid. Also, as cholesterol generally helps to reduce fluidity, once
removed there becomes large gaps in between the relatively freely moving phospholipids
causing larger molecules to be able to diffuse out, similar to the curcumin, therefore the cell
membrane is increasingly more permeable. The concentration in this beaker is higher than
the previous beaker, so the damage to the cell membrane is worse and much larger gaps
and holes are formed. Thus, more pigments leak out resulting in much brighter coloured
solution.

CONCLUSION

Based on the result, there is a clear relationship between denaturing solvent ( ethanol) and
the effect of ethanol on the structure of the cell membrane of Curcuma longa. Ethanol is a
non-polar solvent and disrupts the structure of cell membrane as it increases membrane
fluidity. The greater the ethanol concentration, the stronger more the effect to the cell
membrane. The ethanol concentration influenced the cell membrane structure as this leads
to more gaps in the phospholipid bilayer and caused membrane to become more permeable
and fluidity. For the final result, the beaker which Curcuma longa in 100% ethanol shows the
brightest colour in the solution compared to others. Therefore, 100% of ethanol has the
overall greatest effect on membrane permeability of Curcuma longa than water and 25%
ethanol.

REFERENCES

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https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1367264/#:~:text=Ethanol%20is%20able
%20to%20form,of%20ethanol%20through%20the%20bilayer

https://www.quora.com/What-is-the-effect-of-ethanol-on-the-beetroot-cell%E2%80%99s-
membrane

https://pubmed.ncbi.nlm.nih.gov/3526990/#:~:text=Abstract,most%20easily%20disordered
%20by%20ethanol.&text=Ethanol%20may%20favor%20the%20uptake,thus%20reducing
%20its%20own%20effect

http://www.markedbyteachers.com/as-and-a-level/science/the-effect-of-ethanol-
concentration-on-the-permeability-of-beetroot-cell-membranes-to-betalain.html

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