passive transport lab experiment

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Egg Osmosis Sample 2 lab

Introduction: Transport can be either passive or active. Passive transport is the movement of substances across the membrane without any input of energy by the cell. Active transport is the movement of materials where a cell is required to expend energy. In the case of this lab the discussion will be centered on passive transport. The simplest type of passive transport is diffusion . Diffusion is the movement of molecules from an area of higher to an area of lower concentration without any energy input. Diffusion is driven by the kinetic energy found in the molecules. Diffusion will eventually cause the concentration of molecules to be the same throughout the space the molecules occupy, causing a state of equilibrium to exist. Another type of passive transport is that of osmosis . Osmosis is the movement of water across a semi-permeable membrane. The process by which osmosis occurs is when water molecules diffuse across a cell membrane from an area of higher concentration to an area of lower concentration. The direction of osmosis depends on the relative concentration of the solutes on the two sides. In osmosis, water can travel in three different ways. If the molecules outside the cell are lower than the concentration in the cytosol, the solution is said to be hypotonic to the cytosol, in this process, water diffuses into the cell until equilibrium is established. If the molecules outside the cell are higher than the concentration in the cytosol, the solution is said to be hypertonic to the cytosol, in this process, water diffuses out of the cell until equilibrium exists. If the molecules outside and inside the cell are equal, the solution is said to be isotonic to the cytosol, in this process, water diffuses into and out of the cell at equal rates, causing no net movement of water. In osmosis the cell is selectively permeable , meaning that it only allows certain substances to be transferred into and out of the cell. In osmosis, the proteins only on the surface are called peripheral proteins, which form carbohydrate chains whose purpose is used like antennae for communication. Embedded in the peripheral proteins are integral proteins that can either be solid or have a pore called channel proteins . Channel proteins allow glucose, or food that all living things need to live, pass through.

Hypothesis: In the syrup solution, there will be a net movement of molecules out of the egg, and in the water solution, the molecules will diffuse in and out of the cell at equal rates.

Materials: The materials used in this lab were 2 fresh eggs in the shell, an overhead marker, 400 ml of water, graduated cylinder, 1 large beaker, 2 medium beakers, 1 small beaker, white vinegar, Karo syrup, distilled water, pencil, paper, lab apron, lab goggles, saran wrap, masking tape, plastic tray, tongs, electronic balance, osmosis lab sheet, and computer.

Methods: On day 1 , measure the masses of both the eggs with the shell. Label 1 beaker vinegar , and then use the graduated cylinder to measure 400 mL of vinegar to put in the labeled beaker. Place both eggs in the solution (place a small beaker on top of the eggs, if necessary) then cover. Let the eggs stand for 24 hours or more to remove the shell.

On day 2 , record the observations of what happened to the eggs in the vinegar solution. Carefully, remove the eggs from the vinegar, gently rinsing the eggs off in water. Clean the beakers used for the vinegar solution preparing them for the syrup solution, and then label the 2 medium beakers syrup. Before the eggs are placed in the syrup solution record the mass of both eggs then put it on the datasheet. After that has been done, place the eggs in the beaker, pouring enough syrup to cover the eggs, cover them loosely and let them stand for 24 hours.

On day 3 , record the observations of the egg from the syrup solution. Carefully, remove the eggs from the beakers, gently rinsing the syrup off of the eggs. Pour the remaining syrup in the container provided by the teacher. Clean the two beakers used in the syrup solution, preparing them for the water solution. Before the eggs are placed in the water solution record the mass of both eggs then put it on the datasheet. After that has been done, using a graduated cylinder, measure out 200 mL of water for each beaker. Place the eggs in the water solution, cover and let stand 24 hours.

On day 4 , record the observations of the egg from the water solution. Carefully remove the eggs from the beakers, gently rinsing them off. Mass both of the eggs. After the teacher has came and looked at the eggs, discard in the proper place.

Table 1- Egg 1 Data

Table 2- Egg 2 Data

1. When the egg was placed in the water in which direction did the water molecules move? The water molecules moved in the egg.

2. On what evidence do you base this? The molecules moved in, because the size of the egg increased

3. How do you explain the volume of liquid remaining when the egg was removed from the syrup? Since, the cell is selectively permeable, it only allowed a certain amount of the syrup to be present in the cell, just enough to shrink it and also equilibrium was reached..

4. When the egg was placed in the water after being removed from the syrup in which direction did the water move? The water moved in.

5. Why did the water molecules travel better inside the cell than the syrup molecules? The water molecules traveled better into the cell because smaller molecules travel better than other larger molecules.

6. What was the purpose of placing the egg in vinegar? The vinegar solution was only used to remove the shell off the egg.

Error Analysis: A possible error in this lab occurred by having to leave the egg in vinegar for two days instead of one to remove the shell. This caused the egg to initially take in more water.

Discussion and Conclusion: Based on the data collected and the results of the experiment, the hypothesis was correct. The egg appeared shriveled after removing it from the syrup because of the movement of water out of the egg. The syrup solution was hypertonic so water moved out of the egg from an area where water was more concentrated to the outside of the egg where water was less concentrated due to the high amount of sugar or solute. The acetic acid in vinegar did remove the shell from the egg, because the egg required two days to completely remove the shell, some water did move into the egg causing its initial mass without the shell to be higher than the egg’s mass with its shell. Whenever the egg was transferred from the syrup to the distilled water, the concentration of water outside the shriveled egg was greater than the water concentration inside the egg; therefore, water moved into the egg until equilibrium was reached. At that point, movement into and out of the egg continued with no net movement of water molecules. Additional research to see if the egg would have went through cytolysis in another 24 or more hours in the water solution would have been interesting.

5.2 Passive Transport

Learning objectives.

By the end of this section, you will be able to do the following:

Explain why and how passive transport occurs
Understand the osmosis and diffusion processes
Define tonicity and its relevance to passive transport

Plasma membranes must allow certain substances to enter and leave a cell, and prevent some harmful materials from entering and some essential materials from leaving. In other words, plasma membranes are selectively permeable (semipermeable)—they allow some substances to pass through, but not others. If they were to lose this selectivity, the cell would no longer be able to sustain itself, and it would be destroyed. Some cells require larger amounts of specific substances. They must have a way of obtaining these materials from extracellular fluids. This may happen passively, as certain materials move back and forth, or the cell may have special mechanisms that facilitate transport. Some materials are so important to a cell that it spends some of its energy, hydrolyzing adenosine triphosphate (ATP), to obtain these materials. Red blood cells use some of their energy doing just that. Most cells spend the majority of their energy to maintain an imbalance of sodium and potassium ions between the cell's interior and exterior, as well as on protein synthesis.

The most direct forms of membrane transport are passive. Passive transport is a naturally occurring phenomenon and does not require the cell to exert any of its energy to accomplish the movement. In passive transport, substances move from an area of higher concentration to an area of lower concentration. A physical space in which there is a single substance concentration range has a concentration gradient .

Selective Permeability

Plasma membranes are asymmetric: the membrane's interior is not identical to its exterior. There is a considerable difference between the array of phospholipids and proteins between the two leaflets that form a membrane. On the membrane's interior, some proteins serve to anchor the membrane to cytoskeleton's fibers. There are peripheral proteins on the membrane's exterior that bind extracellular matrix elements. Carbohydrates, attached to lipids or proteins, are also on the plasma membrane's exterior surface. These carbohydrate complexes help the cell bind required substances in the extracellular fluid. This adds considerably to plasma membrane's selective nature ( Figure 5.7 ).

Recall that plasma membranes are amphiphilic: They have hydrophilic and hydrophobic regions. This characteristic helps move some materials through the membrane and hinders the movement of others. Non-polar and lipid-soluble material with a low molecular weight can easily slip through the membrane's hydrophobic lipid core. Substances such as the fat-soluble vitamins A, D, E, and K readily pass through the plasma membranes in the digestive tract and other tissues. Fat-soluble drugs and hormones also gain easy entry into cells and readily transport themselves into the body’s tissues and organs. Oxygen and carbon dioxide molecules have no charge and pass through membranes by simple diffusion.

Polar substances present problems for the membrane. While some polar molecules connect easily with the cell's outside, they cannot readily pass through the plasma membrane's lipid core. Additionally, while small ions could easily slip through the spaces in the membrane's mosaic, their charge prevents them from doing so. Ions such as sodium, potassium, calcium, and chloride must have special means of penetrating plasma membranes. Simple sugars and amino acids also need the help of various transmembrane proteins (channels) to transport themselves across plasma membranes.

Diffusion is a passive process of transport. A single substance moves from a high concentration to a low concentration area until the concentration is equal across a space. You are familiar with diffusion of substances through the air. For example, think about someone opening a bottle of ammonia in a room filled with people. The ammonia gas is at its highest concentration in the bottle. Its lowest concentration is at the room's edges. The ammonia vapor will diffuse, or spread away, from the bottle, and gradually, increasingly more people will smell the ammonia as it spreads. Materials move within the cell’s cytosol by diffusion, and certain materials move through the plasma membrane by diffusion ( Figure 5.8 ). Diffusion expends no energy. On the contrary, concentration gradients are a form of potential energy, which dissipates as the gradient is eliminated.

Each separate substance in a medium, such as the extracellular fluid, has its own concentration gradient, independent of other materials' concentration gradients. In addition, each substance will diffuse according to that gradient. Within a system, there will be different diffusion rates of various substances in the medium.

Factors That Affect Diffusion

Molecules move constantly in a random manner, at a rate that depends on their mass, their environment, and the amount of thermal energy they possess, which in turn is a function of temperature. This movement accounts for molecule diffusion through whatever medium in which they are localized. A substance moves into any space available to it until it evenly distributes itself throughout. After a substance has diffused completely through a space, removing its concentration gradient, molecules will still move around in the space, but there will be no net movement of the number of molecules from one area to another. We call this lack of a concentration gradient in which the substance has no net movement dynamic equilibrium. While diffusion will go forward in the presence of a substance's concentration gradient, several factors affect the diffusion rate.

Extent of the concentration gradient: The greater the difference in concentration, the more rapid the diffusion. The closer the distribution of the material gets to equilibrium, the slower the diffusion rate.
Mass of the molecules diffusing: Heavier molecules move more slowly; therefore, they diffuse more slowly. The reverse is true for lighter molecules.
Temperature: Higher temperatures increase the energy and therefore the molecules' movement, increasing the diffusion rate. Lower temperatures decrease the molecules' energy, thus decreasing the diffusion rate.
Solvent density: As the density of a solvent increases, the diffusion rate decreases. The molecules slow down because they have a more difficult time passing through the denser medium. If the medium is less dense, diffusion increases. Because cells primarily use diffusion to move materials within the cytoplasm, any increase in the cytoplasm’s density will inhibit the movement of the materials. An example of this is a person experiencing dehydration. As the body’s cells lose water, the diffusion rate decreases in the cytoplasm, and the cells’ functions deteriorate. Neurons tend to be very sensitive to this effect. Dehydration frequently leads to unconsciousness and possibly coma because of the decrease in diffusion rate within the cells.
Solubility: As we discussed earlier, nonpolar or lipid-soluble materials pass through plasma membranes more easily than polar materials, allowing a faster diffusion rate.
Surface area and plasma membrane thickness: Increased surface area increases the diffusion rate; whereas, a thicker membrane reduces it.
Distance travelled: The greater the distance that a substance must travel, the slower the diffusion rate. This places an upper limitation on cell size. A large, spherical cell will die because nutrients or waste cannot reach or leave the cell's center, respectively. Therefore, cells must either be small in size, as in the case of many prokaryotes, or be flattened, as with many single-celled eukaryotes.

A variation of diffusion is the process of filtration. In filtration, material moves according to its concentration gradient through a membrane. Sometimes pressure enhances the diffusion rate, causing the substances to filter more rapidly. This occurs in the kidney, where blood pressure forces large amounts of water and accompanying dissolved substances, or solutes , out of the blood and into the renal tubules. The diffusion rate in this instance is almost totally dependent on pressure. One of the effects of high blood pressure is the appearance of protein in the urine, which abnormally high pressure "squeezes through".

Facilitated transport

In facilitated transport , or facilitated diffusion, materials diffuse across the plasma membrane with the help of membrane proteins. A concentration gradient exists that would allow these materials to diffuse into the cell without expending cellular energy. However, these materials are polar molecule ions that the cell membrane's hydrophobic parts repel. Facilitated transport proteins shield these materials from the membrane's repulsive force, allowing them to diffuse into the cell.

The transported material first attaches to protein or glycoprotein receptors on the plasma membrane's exterior surface. This allows removal of material from the extracellular fluid that the cell needs. The substances then pass to specific integral proteins that facilitate their passage. Some of these integral proteins are collections of beta-pleated sheets that form a pore or channel through the phospholipid bilayer. Others are carrier proteins which bind with the substance and aid its diffusion through the membrane.

The integral proteins involved in facilitated transport are transport proteins , and they function as either channels for the material or carriers. In both cases, they are transmembrane proteins. Channels are specific for the transported substance. Channel proteins have hydrophilic domains exposed to the intracellular and extracellular fluids. In addition, they have a hydrophilic channel through their core that provides a hydrated opening through the membrane layers ( Figure 5.9 ). Passage through the channel allows polar compounds to avoid the plasma membrane's nonpolar central layer that would otherwise slow or prevent their entry into the cell. Aquaporins are channel proteins that allow water to pass through the membrane at a very high rate.

Channel proteins are either open at all times or they are “gated,” which controls the channel's opening. When a particular ion attaches to the channel protein it may control the opening, or other mechanisms or substances may be involved. In some tissues, sodium and chloride ions pass freely through open channels; whereas, in other tissues a gate must open to allow passage. An example of this occurs in the kidney, where there are both channel forms in different parts of the renal tubules. Cells involved in transmitting electrical impulses, such as nerve and muscle cells, have gated channels for sodium, potassium, and calcium in their membranes. Opening and closing these channels changes the relative concentrations on opposing sides of the membrane of these ions, resulting in facilitating electrical transmission along membranes (in the case of nerve cells) or in muscle contraction (in the case of muscle cells).

Carrier Proteins

Another type of protein embedded in the plasma membrane is a carrier protein . This aptly named protein binds a substance and, thus triggers a change of its own shape, moving the bound molecule from the cell's outside to its interior ( Figure 5.10 ); depending on the gradient, the material may move in the opposite direction. Carrier proteins are typically specific for a single substance. This selectivity adds to the plasma membrane's overall selectivity. Scientists poorly understand the exact mechanism for the change of shape. Proteins can change shape when their hydrogen bonds are affected, but this may not fully explain this mechanism. Each carrier protein is specific to one substance, and there are a finite number of these proteins in any membrane. This can cause problems in transporting enough material for the cell to function properly. When all of the proteins are bound to their ligands, they are saturated and the rate of transport is at its maximum. Increasing the concentration gradient at this point will not result in an increased transport rate.

An example of this process occurs in the kidney. In one part, the kidney filters glucose, water, salts, ions, and amino acids that the body requires. This filtrate, which includes glucose, then reabsorbs in another part of the kidney. Because there are only a finite number of carrier proteins for glucose, if more glucose is present than the proteins can handle, the excess is not transported and the body excretes this through urine. In a diabetic individual, the term is “spilling glucose into the urine.” A different group of carrier proteins, glucose transport proteins, or GLUTs, are involved in transporting glucose and other hexose sugars through plasma membranes within the body.

Channel and carrier proteins transport material at different rates. Channel proteins transport much more quickly than carrier proteins. Channel proteins facilitate diffusion at a rate of tens of millions of molecules per second; whereas, carrier proteins work at a rate of a thousand to a million molecules per second.

Osmosis is the movement of free water molecules through a semipermeable membrane according to the water's concentration gradient across the membrane, which is inversely proportional to the solutes' concentration. While diffusion transports material across membranes and within cells, osmosis transports only water across a membrane and the membrane limits the solutes' diffusion in the water. Not surprisingly, the aquaporins that facilitate water movement play a large role in osmosis, most prominently in red blood cells and the membranes of kidney tubules.

Osmosis is a special case of diffusion. Water, like other substances, moves from an area of high concentration of free water molecules to one of low free water molecule concentration. An obvious question is what makes water move at all? Imagine a beaker with a semipermeable membrane separating the two sides or halves ( Figure 5.11 ). On both sides of the membrane the water level is the same, but there are different dissolved substance concentrations, or solute , that cannot cross the membrane (otherwise the solute crossing the membrane would balance concentrations on each side). If the solution's volume on both sides of the membrane is the same, but the solute's concentrations are different, then there are different amounts of water, the solvent, on either side of the membrane.

To illustrate this, imagine two full water glasses. One has a single teaspoon of sugar in it; whereas, the second one contains one-quarter cup of sugar. If the total volume of the solutions in both cups is the same, which cup contains more water? Because the large sugar amount in the second cup takes up much more space than the teaspoon of sugar in the first cup, the first cup has more water in it.

Returning to the beaker example, recall that it has a solute mixture on either side of the membrane. A principle of diffusion is that the molecules move around and will spread evenly throughout the medium if they can. However, only the material capable of getting through the membrane will diffuse through it. In this example, the solute cannot diffuse through the membrane, but the water can. Water has a concentration gradient in this system. Thus, water will diffuse down its concentration gradient, crossing the membrane to the side where it is less concentrated. This diffusion of water through the membrane—osmosis—will continue until the water's concentration gradient goes to zero or until the water's hydrostatic pressure balances the osmotic pressure. Osmosis proceeds constantly in living systems.

Tonicity describes how an extracellular solution can change a cell's volume by affecting osmosis. A solution's tonicity often directly correlates with the solution's osmolarity. Osmolarity describes the solution's total solute concentration. A solution with low osmolarity has a greater number of water molecules relative to the number of solute particles. A solution with high osmolarity has fewer water molecules with respect to solute particles. In a situation in which a membrane permeable to water, though not to the solute separates two different osmolarities, water will move from the membrane's side with lower osmolarity (and more water) to the side with higher osmolarity (and less water). This effect makes sense if you remember that the solute cannot move across the membrane, and thus the only component in the system that can move—the water—moves along its own concentration gradient. An important distinction that concerns living systems is that osmolarity measures the number of particles (which may be molecules) in a solution. Therefore, a solution that is cloudy with cells may have a lower osmolarity than a solution that is clear, if the second solution contains more dissolved molecules than there are cells.

Hypotonic Solutions

Scientists use three terms—hypotonic, isotonic, and hypertonic—to relate the cell's osmolarity to the extracellular fluid's osmolarity that contains the cells. In a hypotonic situation, the extracellular fluid has lower osmolarity than the fluid inside the cell, and water enters the cell. (In living systems, the point of reference is always the cytoplasm, so the prefix hypo - means that the extracellular fluid has a lower solute concentration, or a lower osmolarity, than the cell cytoplasm.) It also means that the extracellular fluid has a higher water concentration in the solution than does the cell. In this situation, water will follow its concentration gradient and enter the cell.

Hypertonic Solutions

As for a hypertonic solution, the prefix hyper - refers to the extracellular fluid having a higher osmolarity than the cell’s cytoplasm; therefore, the fluid contains less water than the cell does. Because the cell has a relatively higher water concentration, water will leave the cell.

Isotonic Solutions

In an isotonic solution, the extracellular fluid has the same osmolarity as the cell. If the cell's osmolarity matches that of the extracellular fluid, there will be no net movement of water into or out of the cell, although water will still move in and out. Blood cells and plant cells in hypertonic, isotonic, and hypotonic solutions take on characteristic appearances ( Figure 5.12 ).

Visual Connection

A doctor injects a patient with what the doctor thinks is an isotonic saline solution. The patient dies, and an autopsy reveals that many red blood cells have been destroyed. Do you think the solution the doctor injected was really isotonic?

Link to Learning

For a video illustrating the diffusion process in solutions, visit this site .

Tonicity in Living Systems

In a hypotonic environment, water enters a cell, and the cell swells. In an isotonic condition, the relative solute and solvent concentrations are equal on both membrane sides. There is no net water movement; therefore, there is no change in the cell's size. In a hypertonic solution, water leaves a cell and the cell shrinks. If either the hypo- or hyper- condition goes to excess, the cell’s functions become compromised, and the cell may be destroyed.

A red blood cell will burst, or lyse, when it swells beyond the plasma membrane’s capability to expand. Remember, the membrane resembles a mosaic, with discrete spaces between the molecules comprising it. If the cell swells, and the spaces between the lipids and proteins become too large, the cell will break apart.

In contrast, when excessive water amounts leave a red blood cell, the cell shrinks, or crenates. This has the effect of concentrating the solutes left in the cell, making the cytosol denser and interfering with diffusion within the cell. The cell’s ability to function will be compromised and may also result in the cell's death.

Various living things have ways of controlling the effects of osmosis—a mechanism we call osmoregulation. Some organisms, such as plants, fungi, bacteria, and some protists, have cell walls that surround the plasma membrane and prevent cell lysis in a hypotonic solution. The plasma membrane can only expand to the cell wall's limit, so the cell will not lyse. The cytoplasm in plants is always slightly hypertonic to the cellular environment, and water will always enter a cell if water is available. This water inflow produces turgor pressure, which stiffens the plant's cell walls ( Figure 5.13 ). In nonwoody plants, turgor pressure supports the plant. Conversely, if you do not water the plant, the extracellular fluid will become hypertonic, causing water to leave the cell. In this condition, the cell does not shrink because the cell wall is not flexible. However, the cell membrane detaches from the wall and constricts the cytoplasm. We call this plasmolysis . Plants lose turgor pressure in this condition and wilt ( Figure 5.14 ).

Tonicity is a concern for all living things. For example, paramecia and amoebas, which are protists that lack cell walls, have contractile vacuoles. This vesicle collects excess water from the cell and pumps it out, keeping the cell from lysing as it takes on water from its environment ( Figure 5.15 ).

Many marine invertebrates have internal salt levels matched to their environments, making them isotonic with the water in which they live. Fish, however, must spend approximately five percent of their metabolic energy maintaining osmotic homeostasis. Freshwater fish live in an environment that is hypotonic to their cells. These fish actively take in salt through their gills and excrete diluted urine to rid themselves of excess water. Saltwater fish live in the reverse environment, which is hypertonic to their cells, and they secrete salt through their gills and excrete highly concentrated urine.

In vertebrates, the kidneys regulate the water amount in the body. Osmoreceptors are specialized cells in the brain that monitor solute concentration in the blood. If the solute levels increase beyond a certain range, a hormone releases that slows water loss through the kidney and dilutes the blood to safer levels. Animals also have high albumin concentrations, which the liver produces, in their blood. This protein is too large to pass easily through plasma membranes and is a major factor in controlling the osmotic pressures applied to tissues.

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Access for free at https://openstax.org/books/biology-2e/pages/1-introduction

Authors: Mary Ann Clark, Matthew Douglas, Jung Choi
Publisher/website: OpenStax
Book title: Biology 2e
Publication date: Mar 28, 2018
Location: Houston, Texas
Book URL: https://openstax.org/books/biology-2e/pages/1-introduction
Section URL: https://openstax.org/books/biology-2e/pages/5-2-passive-transport

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