BIOL160 Lab Report 4

Movement In And Out of the Cells

By

: Liu Ming Jen [1102H12720]

Lecturer

: Ms. Bessie Ong

Experiment Date : March 14th, 2012

Introduction There are two ways that molecules can move across the plasma membrane which is active transport and passive transport. Active transport is the movement of molecules or ions from a region of lower concentration to a region of higher concentration (against the concentration gradient) across the plasma membrane, with the use of cellular energy (Gan, 2006, pg.63). Passive transport is the movement of

substances in cells which does not require energy. Passive transport is divided into three: simple diffusion, facilitated diffusion and osmosis. Simple diffusion is the net movement of molecules or ions from a region of high concentration to a region of lower concentration (according the concentration gradient). Simple diffusion is a physical process occurs in liquids or gases. Lipid soluble molecules, water, oxygen and carbon dioxide move across plasma membrane by simple diffusion (Gan, 2006, pg.57). Facilitated diffusion is the movement of hydrophilic molecules or ions across the plasma membrane with the help of transport proteins. The transport proteins called carrier proteins have binding sites that can combine reversibly with specific molecules and transport them across the plasma membrane. The carrier proteins are specific and can only combine with certain molecules. Carrier protein enables macromolecules to move across the plasma membrane at a faster rate (Gan, 2006, pg.62). Osmosis is the net movement of water molecules from a region of low solute concentration to a region of high solute concentration through a semi-permeable membrane. Semi-permeable membranes are very thin layers of material such as cell membranes which allow some things to pass through them but prevent other things from passing through. Cell membranes will allow small molecules like oxygen, water, carbon dioxide, ammonia, glucose and amino-acids to pass through. Cell membranes will not allow larger molecules like sucrose, starch and protein to pass through (Punchon, 2009). For example, water molecules move across the phospholipid bilayer and pores in the plasma membrane by the process of osmosis (Gan, 2006, pg.58). The similarity of simple diffusion, facilitated diffusion and osmosis is the molecules move down the concentration gradient and the cell does not required to expand energy. The molecules will continue to move from regions of higher concentration to region of lower concentration until dynamic equilibrium is reached. The molecules are evenly dispersed and the concentration gradient no longer exists in dynamic equilibrium state. Simple diffusion differs from osmosis because simple diffusion involves the movement of any molecules but osmosis only involves movement of water molecules. Besides, osmosis only occurs through a semipermeable membrane while simple diffusion does not require a semi-permeable membrane. Facilitated diffusion is different from both simple diffusion and osmosis because the molecules in facilitated diffusion diffuse passively into the cell only with the help of transport proteins (Gan, 2006, pg.60). Hypotonic solutions are the solutions that contain a higher concentration of water than the cytoplasm. A hypotonic environment is created when the solution outside of the cell is hypotonic in comparison to the cytoplasm of the cell. For example, the cell has 5% solute and 95% water while outside the cell is distilled water which has 0% solute and 100% water. In this situation, water outside of the cell will move by osmosis to the inside of the cell in an attempt to equalize the concentrations (LearnNowBC, 2009). The cell gains water and this will cause swelling and an

increase in internal pressure. This may cause lysis in animal cell and the cell may eventually burst. This condition that happens to red blood cell is known as haemolysis. In plant cell, the cell is said to be turgid and firm. The plant cell does not burst because the rigid cell wall is strong enough to resist the high pressure inside the cell (Gan, 2006, pg.67). Hypertonic solutions contain a higher concentration of solute than the cell. A hypertonic environment is created when the solution outside of the cell is hypertonic in comparison to the cytoplasm of the cell. For example, the cell has 1% solute and 99% water while the solution outside the cell has 5% solute and 95% water. Since the concentration of water is higher inside the cell, there is a net movement of water by osmosis from inside to the outside of the cell in an attempt to equalize the concentrations. As a result, water leaves the cell. This causes shrinkage of the cell as its internal pressure decreases. An animal cell that shrunk in this manner is said to be crenated (LearnNowBC, 2009). While in plant cell, this phenomenon is called plasmolysis which is the shrinking of the cytoplasm due to osmosis (Gan, 2006, pg.68). Isotonic solutions contain the same concentration of dissolved solute as the cell. An isotonic environment is created when the solution outside of the cell is isotonic with the cytoplasm of the cell. In this situation, the solution outside of the cell and the cytoplasm of the cell would both be 1% solute and 99% water. Water is found to have equal concentrations both inside and outside of the cell (LearnNowBC, 2009). Water diffuses in and out of the cell at equal rates therefore there is no net movement of water or change in cell volume. Both animal and plant cells retain their normal shape. Hypothesis Experiment a): If the rate of diffusion of the molecules pass the permeable membrane is related to the size of molecules, then the percentage of glucose molecuses diffused would be more than the potassium iodide molecules Experiment b): If the effect of plasmolysis in the onion cell is related to the concentration of sucrose solution, then the onion cell immersed in the 2.0M sucrose solution will have higher plasmolysis than the one in 1.0M sucrose solution.

Material and Method A) Movement through an artificial semi-permeable membrane A dialysis bag was prepared by folding over 1 2 cm of one end of a piece of dialysis tubing that had been soaked in water for few minutes. The folded end was tied with a rubber band, forming a bag. The opposite end of the bag was rolled between fingers until it was opened and the bag was half filled with 10% glucose solution. Then an equal amount of starch solution was added into the bag. The bag was held closed and its contents were mixed. The color of the mixture was recorded. The outside of the bag was carefully rinsed in tap water. A small beaker was half filled with water and several drops of KI-iodine solution were added to the water until it is visibly yellow. The bag was placed in the beaker so that the untied end of the bag was hung over the edge of the beaker. The mixture was not allowed to spill out of the bag. The bag was left in the beaker for about 30 minutes. After 30 min, the bag was carefully removed and the color changes were recorded. Benedicts test was performed for the presence of reducing sugar in the solution. About 3 cm3 of the mixture in the dialysis bag was removed into a test tube. Similarly about 3 cm3 of KIiodine mixture in the beaker was moved into a separate test tube. Both test tubes were then labeled. Half cm3 of Benedicts solution was added to each tube. The test tubes were heated in boiling water for about 10 min and the results were recorded.

B) Movement through a cell membrane One of the fleshy scale leaves of an onion was removed. With a sharp razor blade, the inner epidermis was cut into three squares of approximately 5 mm. The three squares were placed on a slide and each was flooded with distilled water for 5 minutes. After 5 min, the second square was placed on another slide and was flooded with 1.0 M sucrose solution, while the third piece was placed on the third slide and flooded with 2.0 M sucrose solution. The epidermis in the sucrose solution was left for at least 10 min. After 10 min, each piece of epidermis was covered with a cover slip and excess liquid was blotted off before the conditions of the cells were observed under the microscope.

Results A) Movement through an artificial semi-permeable membrane. Table 1: Results for Benedicts test on the solutions Solution Dialysis Beaker Original Colorless yellow Before Benedicts solution is added Light Blue Yellow After Benedicts solution is added Dark blue Blue After being heated Yellow Brick red

B) Movement through a cell membrane Refer to Figure 1.1, 1.2 and 1.3

Discussion A) Movement through an artificial semi-permeable membrane This experiment is basically based on the osmosis that says water diffuses through a selectively permeable membrane from where it has higher concentration of water to where its concentration is lower. In the respect, the aim of this experiment is to testify if two solutions of glucose and starch of different molecular mass are separated by the membrane to pass through the pore of the membrane. (this experiment is specifically to prove that the size of iodine molecules is smaller than the size of starch molecules.) The change of the color from colorless to dark blue in the dialysis bag proves that the KI-iodine molecules in the beaker are small enough to pass through the selectively permeable tubing and move into dialysis bag. In other words, KI-iodine molecules are able to diffuse through semi permeable membrane and react with starch. However, the color of the solution in the beaker remains the same after being experimented and it directly means that the starch molecules are too large to diffuse through the pores on the membrane of the bag compare to iodine molecules. After the Benedicts test is performed, it is proven that glucose molecules can easily diffuse through the membrane of the bag and enter the solution in the beaker as glucose molecules are small enough to pass through. The color change of the mixture to yellowish brown precipitate has shown the presence of glucose molecules. I also predict that glucose molecules are smaller in size if compared with KI molecules. This is because the KI molecules pass through the bag in a relatively slow rate unlike the glucose molecules that pass through the bag into the solution in the beaker in a faster rate as the size of the solute is highly correlated with its molecular weight. The heavier, larger solute moves more slowly along the concentration gradient than smaller lighter solutes. Thus, dialysis is most effective in removing small solutes and less effective in removing larger solutes. After being left for 30 minutes, dark blue precipitates were formed in the initial pure milky white mixture of glucose and starch solutions in the dialysis bag. It is predicted that the KI-iodine molecules moved through the dialysis bag from the beaker water and reacted with starch in the dialysis bag. KI-iodine turns from yellow to dark blue in the presence of starch a type of polysaccharide. However, the beaker water remained yellow, indicating no starch is present outside the bag. This showed that starch did not diffuse into the water through the semi-permeable dialysis bag. Once investigating the presence of starch in the two mixtures (inside and outside the dialysis bag) had been carried out, Benedicts test followed to see if glucose is present in both mixtures. The mixture from the dialysis bag turned dirty yellow and that taken from the beaker water turned light brick red after boiling. It is recalled that in Benedicts test, sample turns brick red in the presence of glucose. The results indicated that excess glucose solution flowed out from the dialysis bag into the beaker of water diffusion of glucose molecules through the membrane occurred. The difference in color showed that the concentration of glucose molecules is relatively higher in the beaker water. Since starch molecules are too large to diffuse through the pores of dialysis bag, only glucose molecules can do a part in lowering the concentration gradient between the mixtures in the bag and beaker. In order to even out the concentrations, more glucose molecules had diffused into the beaker. As

a result, the color of the mixture in the beaker (light brick red) turned more significantly as compared to the one in the tube (dirty yellow).

B) Movement through a cell membrane The distilled water is hypotonic to the cell sap of the epidermal cells, which means the net inflow of water by osmosis into the vacuoles causes them to swell. It means the cells are highly turgid at this point. The vacuoles and cytoplasm exert an outward force against the plasma membrane and cell wall but the cell do not burst because of the rigid cell walls. For the cell immerse in the 1M sucrose solution. The cell is isotonic to the cell sap of the epidermal cells which means the cells do not lose of gain water. As for the cell immerse in the 2M sucrose solution, the cell is hypertonic to the cell sap of the epidermal cells. This means there is a net outflow of water from the vacuoles by osmosis. The plant cells are plasmolysed as a result. Plasmolysis is the contraction of cells within plants due to the loss of water through osmosis (Campbell, 2009). It is the cell membrane peeling off of the cell wall and the vacuole collapsing and it occurs when a plant cell's membrane shrinks away from its cell wall. This occurs when water is drawn out of the cell and into the extracellular fluid. The movement of water occurs across the membrane flowing from a region of high water concentration to a region of low water concentration outside the cell. The cells showed different percentage of plasmolysis in the two sucrose concentration is due to the two sucrose solution gave two different environment (hypotonic, hypertonic) to the cells.

Plasmolysis is a process, which the plasma membrane shrinks and pulls away from the cell wall in a plant cell. Plant wilts if its cells are plasmolyzed. However, plasmolysis only occurs in extreme conditions and rarely happens in nature. In the experiment, only one out of three cells were being plasmolyzed, with another two turned turgid. They all started from turgid condition, as they were all been flooded with distilled water at first. The cells in the experiment showed different % of plasmolysis in the two sucrose solutions due to the different osmotic pressure outside the cell. In higher concentrated solution (also called hypertonic solution), the external osmosis pressure is higher. This higher external osmotic pressure causes more water to flow out of the cell (immersed in 2.0 M sucrose solution) and results in greater shrinkage of the plasma membrane (pulling more away from cell wall) as compared to the cell placed in 1.0 M of sucrose solution. In these hypertonic solutions, the plant cells were weakened and looked very unhealthy. As for the cells flooded with only distilled water (hypotonic solution), they looked very healthy and pretty. Despite the high osmotic pressure in the cell, which caused excessive amount of water to enter it, the strong cell wall provided the shape and prevented the plant cell from bursting. The protoplasm of the cell is stretched and touching the cell wall, giving the cell its turgid characteristics.

In conclusion, glucose molecules are small enough to diffuse through the pores on the dialysis bag. However, starch molecules proved too large to diffuse through the pores. Diffusion (spreading out of particles) of glucose occurred due to high concentration gradient high inside the bag but low outside the bag. On the other hand, high osmotic pressure will force water to move to the more concentrated region through osmosis.

Work Cited

Cool School Online Content Development. LearnNowBC .Hypertonic, Isotonic, and Hypotonic Environments. http://www.coolschool.ca/lor/BI12/unit4/U04L06.htm. Retrieved on 19th, March 2012. Gan, W.Y. 2006. Exploring Biology. Shah Alam. Pernerbit Fajar Bakti Sdn. Bhd. Pg57-68. Osmosis. Nigel.D.Purchon. Explanation. http://www.purchon.com/biology/osmosis.htm. Retrieved on 19th, March 2012. Campbell, N.A. and Reece, J.B. and Simon, E.J. 2007. Essential Biology. Third Edition. San Francisco: PEARSON, Benjamin Cummings. p38-42.