movement of substance through cell membrane experiment demonstrating diffusion

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Exp. 1 Movement of substance through cell membrane

Hospitality (bshtm), western philippines university.

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Name: _____________________________ Date Performed: ____________ Year and Course: ____________________ Date Submitted: ____________ Subject title: ________________________ Code number: _______________ Name of Professor: ___________________ Score: ______________________

Exercise no. 1 Movement of Substance through Cell Membrane Experiment Demonstrating Diffusion

I. INTRODUCTION

If we put a teaspoon of instant ice tea on the surface of a glass of water, the molecules soon spread throughout the solution. The molecules of both the solute (ice tea) and the solvent (water) are propelled by random molecular motion. The initially concentrated tea becomes more and more dilute. This process of the net movement of a solute with the gradient (from an area of high concentration to an area of low concentration) is called diffusion.

If the solute can pass through the membrane, diffusion will occur with net transport of material from the region of initial high concentration to the region of initial low concentration, and substance will equilibrate across the cell membrane. After a while, the concentration of the substance will be the same on both sides of the membrane; the system will be at equilibrium, and no more net change will occur.

II. OBJECTIVES At the end of the experiment, each student will be able to:

III. MATERIALS:

2 – 250ml beaker, 1-500ml beaker, 1-1000ml beaker, 5 test tubes, test tube rack, test tube brush, test tube holder, 3 x10 cellophane bag, 1-rubber band, iron ring, 3 pipette, 3-medicine dropper, water bath, electric stove, 1-25ml graduated cylinder, 1m string, ruler

5g powder gelatin, 1ml methylene blue, 2g KMnO4, 2 g NaCl, 5% glucose soln, 2g albumin powder, 2ml Nitric acid, 2ml silver nitrate, 2ml Benedict’s soln.

IV. PROCEDURE:

Movement of dye through the gel

Preliminary preparation ( a day or two before the laboratory period ): add 5 gram (or 1 tsp) of gelatin to 25ml of cold water. Let stand for 5 minutes. Add 75 ml of boiling water and stir until dissolved. Pour about 15 ml of the solution into a test tube. Fill the remaining solution to another test tube refrigerate until gelled.

Place a drop of methylene blue on the surface of the gel prepared ahead of time. Set aside at room temperature. At the end of 1 hour, 2 hours, 6 hours,12 hours and 24 hours, observe whether the dye has move and if so in which direction and how far.

Diffusion of KMnO 4

• Place a few crystal of potassium permanganate on the bottom of a beaker half filled with water. Observe.

The dialyzing membrane

Prepare a bag of 3 x 10 inch of cellophane

Prepare a solution as follows in a large beaker (500ml): to 100ml of water add ½ tsp of NaCI plus 50ml of 5% glucose, and the uncooked albumin and place them inside the cellophane bag. Both NaCI and glucose are crystalloids or true solutes, whereas albumin is a colloidal solute. ( Better look at these terms up if you are not sure what they mean)

Tie a string around the top of the bag and suspend it in a 1000ml beaker with distilled water. You may use an iron ring and stand for this set-up. If the cellophane bag is very thick you may pinch a very small hole in the bag and suspend it in the distilled water. Let it stand for about an hour and then the test indicated in steps a, b, and c.

a. Test for albumin: Using a pipette, pour about 5 ml of the fluid into the test tube. Add a few drops of nitric acid. Note whether coagulation occurs. Nitric acid coagulates albumin

__________________________________________________________________

• Did the result in step 7, 8 and 9 indicate that the crystalloids glucose and NaCl diffused through the dialyzing membrane? Why?
• Did the colloidal solute albumin diffuse through the membrane in this experiment?
• From the experiments you performed in steps 2 and 3, explain the net diffusion of solutes to its concentration.
• Give at least three (3) factors that affect the movement of materials into the cell through the cell membrane.

VI. CONCLUSION

• Multiple Choice

Course : Hospitality (BSHTM)

University : western philippines university.

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• A-Level Biology Revision Notes >
• CIE A-level Biology Revision Notes

Investigating Transport Across Membranes (A-level Biology)

Investigating transport across membranes, investigating diffusion.

We can investigate how diffusion occurs in biological cells by using cubes of agar jelly. The basic concept of this experiment is outlined below:

Table of Contents

• The agar jelly contains a pH indicator. We can make up agar jelly with an alkaline solution (e.g. sodium hydroxide) and add a few drops of phenolphthalein to it before the jelly sets. Phenolphthalein is a pH indicator which turns pink in the presence of alkaline solutions, thus, the jelly will have a bright pink colour.
• The agar jelly is placed in an acidic solution. Once the jelly has set, we can cut it up into cubes and place it in an acidic solution, such as dilute hydrochloric acid.
• The agar jelly is neutralised by the diffusion of the acid. The acidic solution will slowly diffuse into the agar jelly and neutralise the alkaline solution. As it does, the jelly will lose its pink colour and become colourless, as phenolphthalein turns colourless in non-alkaline environments.

We can alter different parts of this experiment to model how different factors affect the rate of diffusion.

Investigating the effects of surface area on diffusion

• Cut the agar jelly into different sized cubes to investigate the effects of surface area . Cut the jelly into cubes of different sizes and work out each cube’s surface area to volume ratio . For example, a cube with 2cm edges will have a surface area to volume ratio of 3:1.
• Place the cubes in the same volume and concentration of acid. Put the cubes into containers which hold the same volume and concentration of hydrochloric acid. Then measure the time it takes for the different cubes to go colourless.
• The cube with the largest surface area: volume ratio will go colourless the quickest. The cube with the largest surface area: volume ratio has the greatest amount of space available for the hydrochloric acid to diffuse into the jelly so it will be neutralised the fastest.

Investigating the effects of concentration on diffusion

• Place the agar jelly cubes in different concentrations of acid. Cut the agar jelly into equal sized cubes and put them in different containers, each with a different concentration of hydrochloric acid. Measure the time it takes for the different cubes to go colourless.
• The cube placed in the highest concentration of acid will go colourless the quickest. The cube placed in the container with the highest concentration will have the greatest concentration of acid being diffused into the jelly per minute. As such, it will go colourless the quickest.

Investigating the effects of temperature on diffusion

• Place the agar jelly cubes in different temperatures. Cut the agar jelly into equal sized cubes and put them in different containers, each with the same concentration of hydrochloric acid. Put the containers in water baths heated to different temperatures. Be careful not to heat the water baths over 65° as the agar jelly will melt.
• The cube placed in the highest temperature of acid will go colourless the quickest. As high temperatures speed up the rate of diffusion, the cube in the hottest container will be neutralised the quickest.

Investigating Osmosis

Osmosis is the movement of water molecules from an area of high water potential to an of low water potential by osmosis. Water potential is determined by the concentration of solutes in the solutions on either side of the cell membrane.

Investigations using plant tissue

This experiment involves placing plant tissue, e.g. potato cylinders, in varying concentrations of sucrose solutions to determine the water potential of the plant tissues.

• Prepare the different concentrations of sucrose solutions . Using distilled water and 1M sucrose solution, prepare a series of dilutions such that you now have 0.0, 0.2, 0.4, 0.6, 0.8 and 1.0M sucrose. Place 5cm 3 of each dilution into separate beakers.
• Prepare equal sized pieces of potato chips. Using a cork borer, cut out 18 pieces of potato chips, all of equal sizes.
• Weigh the mass of the potato chips. Dry the potato chips gently with a paper towel. Divide them into groups of three and weigh each group.
• Place each group of potato chips in each solution . The potato chips should be left in the solutions for a minimum of 20 minutes.   All groups should be left in the solution for the same amount of time.
• Weigh the mass of the potato chips again. Once your desired amount of time has passed, remove the chips from the solutions, and dry them gently using a paper towel. Reweigh each group again.
• Calculate % change in mass. Using the mass of the potato chips before and after being placed in the solution, calculate the % change in its mass.
• Plot the % change in mass on a calibration curve. The calibration curve helps us determine the water potential in the potato sample. Plot the % change in mass against concentration of sucrose solution.   The point at which the curve crosses the x axis is when the sucrose solution is isotonic with the potato samples i.e. the water potential of the sucrose solution is the same as the water potential of the potatoes. At this point, there is no movement of water in or out of the potato. Overall:
• The potato samples in the dilute solutions will have a net increase in mass – the water potential is greater in the potato than in the sucrose solution, so water moves into the potato samples via osmosis.
• The potato samples in the concentrated solutions will have a net decrease in mass – the water potential is lower in the potato than in the sucrose solution, so water moves out of the potato samples via osmosis.

Investigations using Visking tubing

Visking tubing is an artificial membrane that is selectively permeable as it has many microscopic pores. This allows smaller molecules such as water and glucose to pass through it, while larger molecules such as starch and sucrose are unable to cross the membrane.

• Prepare three equal-sized pieces of Visking tubings. Run the tubing under tap water to soften it and knot each tubing on one end to create a bag.
• Place a rubber bung at the open end of the Visking tubing. Find rubber bungs with an opening in the centre that will fit the open end of the Visking tubing. Then seal the tubing using the bung and fix it in place using a rubber band.
• Prepare sucrose solutions with concentrations of 0.5M and 1.0M. You may wish to add a food dye to the 0.5M solution so that it is easier to see later on.
• Pipette in the 0.5M sucrose solution. Using a pipette or a syringe, fill each tubing through the opening of the rubber bung with the 0.5M sucrose solution. Make sure it is filled completely to the brim with no air bubbles.
• Insert capillary tubes into each of the tubings . Insert a capillary tube through the rubber bung’s opening. Mark the level at which the sucrose solution has risen to in the capillary tube.
• Place each Visking tubing into containers of different solutions. Prepare three beakers, each containing distilled water, 0.5M sucrose, and 1.0M sucrose. Place each Visking tubing into each of the beakers and leave them in for the same amount of time.
• Measure the change in liquid level. Mark the new liquid level on the capillary tube before removing the Visking tubing from its beaker. Measure the change in the liquid level. Overall:
• The liquid level of the Visking tubing placed in distilled water will have risen as the sucrose solution in the tubing is hypertonic to the water i.e. the sucrose is more concentrated. Thus, there is net movement of water into the Visking tubing via osmosis.
• The liquid level of the Visking tubing placed in 0.5M sucrose will remain the same as the solution inside the tubing and outside the tubing are isotonic i.e. the solutions are the same concentration.
• The liquid level of the Visking tubing placed in 1.0M sucrose will have decreased as the solution inside the tubing is hypotonic to the solution outside the tubing i.e. the solution inside the tubing is less concentrated.

Transport across membranes is the movement of substances such as ions, molecules, and fluids from one side of a biological membrane to the other. This process is crucial for maintaining cellular homeostasis and allowing cells to exchange materials with their environment.

Investigating transport across membranes is important because it helps us understand the mechanisms by which cells regulate the flow of substances in and out of the cell. This is essential for understanding cellular processes such as metabolic reactions, waste removal, and communication between cells.

There are several methods used to investigate transport across membranes, including: Diffusion experiments to study the movement of substances through the lipid bilayer Osmosis experiments to study the movement of water across a semi-permeable membrane Active transport experiments to study the movement of substances against a concentration gradient with the use of energy Electrochemical experiments to study the movement of ions across the membrane

Factors that can affect transport across membranes include the size of the substance being transported, the charge of the substance, the concentration gradient, and the presence of specific transport proteins.

Transport across membranes can be measured in a variety of ways, including measuring changes in substance concentration, changes in electrical potential, and changes in fluid movement.

The limitations of investigating transport across membranes include the difficulty of obtaining pure and intact biological membranes, the potential for damage to the membrane during experimentation, and the limitations of experimental techniques.

In A-Level Biology, knowledge of transport across membranes can be applied to understand cellular processes such as the movement of nutrients and waste, the regulation of cell volume, and the communication between cells. This knowledge is also important for understanding diseases and disorders related to the malfunction of transport processes, such as cystic fibrosis and diabetes.

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CIE 1 Cell structure

Roles of atp (a-level biology), atp as an energy source (a-level biology), the synthesis and hydrolysis of atp (a-level biology), the structure of atp (a-level biology), magnification and resolution (a-level biology), calculating cell size (a-level biology), studying cells: confocal microscopes (a-level biology), studying cells: electron microscopes (a-level biology), studying cells: light microscopes (a-level biology), life cycle and replication of viruses (a-level biology), cie 10 infectious disease, bacteria, antibiotics, and other medicines (a-level biology), pathogens and infectious diseases (a-level biology), cie 11 immunity, types of immunity and vaccinations (a-level biology), structure and function of antibodies (a-level biology), the adaptive immune response (a-level biology), introduction to the immune system (a-level biology), primary defences against pathogens (a-level biology), cie 12 energy and respiration, anaerobic respiration in mammals, plants and fungi (a-level 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biodiversity calculations (a-level biology), introducing biodiversity (a-level biology), the three domain system (a-level biology), phylogeny and classification (a-level biology), classifying organisms (a-level biology), cie 19 genetic technology, cie 2 biological molecules, properties of water (a-level biology), structure of water (a-level biology), test for lipids and proteins (a-level biology), tests for carbohydrates (a-level biology), protein structures: globular and fibrous proteins (a-level biology), protein structures: tertiary and quaternary structures (a-level biology), protein structures: primary and secondary structures (a-level biology), protein formation (a-level biology), proteins and amino acids: an introduction (a-level biology), phospholipid bilayer (a-level biology), cie 3 enzymes, enzymes: inhibitors (a-level biology), enzymes: rates of reaction (a-level biology), enzymes: intracellular and extracellular forms (a-level biology), enzymes: mechanism of action (a-level 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Experiment Diffusion through Membranes Experiments​

Diffusion through membranes.

Experiment #1A from Advanced Biology with Vernier

Introduction

Diffusion is a process that allows ions or molecules to move from where they are more concentrated to where they are less concentrated. This process accounts for the movement of many small molecules across a cell membrane. Diffusion is the process by which cells acquire food and exchange waste products. Oxygen, for instance, might diffuse in pond water for use by fish and other aquatic animals. When animals use oxygen, more oxygen will diffuse to replace it from the neighboring environment. Waste products released by aquatic animals are diluted by diffusion and dispersed throughout the pond.

It is important to consider how the rate of diffusion of particles might be affected or altered.

• Diffusion may be affected by the steepness of the concentration gradient (the difference between the number of ions or molecules in one region of a substance and that in an adjoining region).
• Diffusion might be affected by other different, neighboring particles. For instance, if oxygen diffuses towards a single-celled pond organism at a certain rate, will that rate be altered if some other molecule suddenly surrounded the organism?

One way to measure the rate of diffusion of ions is to monitor their concentration in solution over a period of time. Since ions are electrically charged, water solutions containing ions will conduct electricity. A Conductivity Probe is capable of monitoring ions in solution. This probe however, will not measure the amount of electrically neutral molecules dissolved in water. Salts, such as sodium chloride, produce ions when they dissolve in water. If you place a salt solution in a container such as dialysis tubing, the salt can travel through the very small holes in the tubing. When dialysis tubing containing a solution of salt ions is placed into a beaker of water, the ions can diffuse out of the tubing and into the surrounding water. In this way, you will be able to measure the diffusion of salts in a solution of water and determine how concentration gradients and the presence of other particles affect the diffusion of the salt across a membrane.

In this experiment, you will

• Use a Conductivity Probe to measure the ionic concentration of various solutions.
• Study the effect of concentration gradients on the rate of diffusion.
• Determine if the diffusion rate for a molecule is affected by the presence of a second molecule.

Sensors and Equipment

This experiment features the following sensors and equipment. Additional equipment may be required.

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This experiment is #1A of Advanced Biology with Vernier . The experiment in the book includes student instructions as well as instructor information for set up, helpful hints, and sample graphs and data.