Osmosis and Diffusion

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1 Osmosis and Diffusion Per bench Osmosis DI Water bottle ml DI water 200 ml 5% Sucrose Solution 150 ml 25% Sucrose Solution (3) Dialysis Tubes (1) Pipette (5 ml) (3) 200 ml Beaker (1) Digital Scale (6) Strings 15 cm Each (1) Pipette pump (1) Weigh boats (1) Sharpie marker Brownian motion (1) Microscope (1) Microscopes slides (1) Cover slips Diffusion (2) 250 ml beakers 10 Drops 1 M (NaOH) 20 Drops of Iodine 3 drops Phenolphthalein (2) Dialysis Tubes (4) 15 cm Strings 10 ml Starch mixture 1 Pipette Effect of Molecular Weight on Diffusion Potassium Permanganate Methyl Orange Malachite Green Methylene Blue (1) Agar Plate* Ruler Per Class 6 DI water Bottles 1.5 L 5% Sucrose Solution 1.5 L 25% Sucrose Solution (30) Pre-cut Dialysis Tubes in water (6) Pipette (5 ml) (18) 250 ml Beaker (6) Digital Scales and weigh boats String (6) Pipette pumps Microscope slides Cover slips (6) Sharpie markers Carmine Red two small flasks with droppers (6) 1 M (NaOH) small dropper bottles (6) Iodine bottles (6) Phenolphthalein bottles (6) Potassium Permanganate (6) Methyl Orange (6) Malachite Green (6) Methylene Blue (6) Agar Plates* (6) Rulers Dicalcified Egg Demonstration (3) Decalcified Eggs (3) 400 ml Beaker DI Water( included above) 250 ml 25% Sucrose Solution (included above) 250 ml 50% Sucrose Solution Dicalcified Egg Demonstration Procedure 1. The instructor will need three student volunteers for this experiment. 2. The initial weight of the eggs will be recorded to the nearest 0.1 gram and be placed in three different solutions: DI water, 25% sucrose, and 50% sucrose. 3. One hour later, the student volunteers will remove the eggs from the labeled beakers, blot them dry with a paper towel, and record the final weight to the nearest 0.1 gram. 0

2 Osmosis and Diffusion Abstract: This lab is composed of instructor demonstrations, as well as student run experiments meant to demonstrate the various principles regarding osmosis and diffusion. For the instructor demonstrations, you will participate in a discussion that covers the concepts critical to understanding the demonstration, and formulate a hypothesis derived from the discussion. You will also utilize common laboratory techniques to explore the effect of solute concentration on the rate of osmosis. You will look at selective permeability of membranes, the effect of molecular weight on diffusion and the effect that different concentrations of solute will have on a membrane. You will also understand terminology such as Brownian Motion, Semi-permeable membrane, isotonic, hypertonic and hypotonic solutions Fig. 1.0 Water (Solvent molecules) follows the salts (solute molecules) Osmosis (25 minutes working on the lab, 1 hour waiting) In this exercise, you will investigate the effect of solute concentration on osmosis. Remember, osmosis is the diffusion of water across a semi-permeable membrane, from a higher concentration of water to a lower concentration of water. It may help you to remember higher concentrations of water are associated with lower concentrations of solute, and vice versa. Solvent Molecules will flow through a semipermeable membrane to a region of higher solute concentration. Fig. 1.1 Pink dots represent Fig 1.1 Solute molecules are represented in pink, solvent molecules (water) are represented in blue. Make three identical cell models by following these directions. 1. Obtain: Three 250 ml beakers and label them 1, 2, and 3 Six pieces of string Three pieces of pre-soaked dialysis tubing 2. In beaker # 1 add 150 ml of distilled water, bag #1 will be placed into it. 1

3 3. In beaker #2 add 150 ml of 5% sucrose solution, bag #2 will be placed into it. 4. In beaker # 3 add 150 ml of 25% sucrose solution, bag #3 will be placed into it. 5. Fold and tie off one end of the dialysis tubing with the strings provided. 6. Open the other end of the dialysis tubing by rolling it between your fingers. 7. Using the pipette fill each of the bags with 10mL of 5% sucrose solution. 8. Fold and tie off the open end with one of the strings; blot off any excess solution, weigh each bag and label them as bag #1(0%), bag #2(5%) and bag #3(25%). Record the initial weights to the nearest 0.1 gram. 9. Place the bags in their respective beakers at the same time or note the respective start times. 10. After one hour, remove the bags, blot them dry and weigh again. Record the final weight to the nearest 0.1 gram. Brownian motion Simple Diffusion is a process in which molecules go through a semipermeable membrane to reach equal concentrations between both areas of the membrane. In the following experiment you will look at molecules vibration caused by Brownian motion. Dye molecules will bump into water molecules making them run randomly and dissipate from one another. 1. Add a drop of carmine red and watch what happens as the dye moves outward from its source. 2. Place cover slip over drop of dye and observe under high power. Look for vibrating bubbles. The goal is to see the dye particles which are large compared to water molecules. The dye particles will appear as vibrating clear bubbles. For some reason, this phenomenon of Brownian motion is easy to observe on some slides and virtually impossible on others, so please share good views with others and be willing to make new preparations, as this is quickly and easily done. Too much or too little dye does not usually work well. 2

4 Diffusion of Substances through a Semipermeable Membrane (15 min working in the lab, 25 minutes waiting) A semipermeable membrane will restrict flow of molecules or ions that go through it. Fig 1.2 Molecules flow from an area of high concentration to an area of low concentration to reach equilibrium in the system, entropy between molecules will dissipate molecules between the semipermeable membranes. 1. Obtain two 250 ml beakers and label them A and B. Four pieces of string Two pieces of pre-soaked dialysis tubing 2. In beaker A add 200 ml of tap water and add 10 drops of the base, 1 M sodium hydroxide (NaOH). Bag A will be placed into beaker A. 3. Fill beaker B with 200 ml of tap water and add 20 drops of iodine. Bag B will be placed into the beaker B. 4. Fill bag A with 10 ml tap water and 3 drops of phenolphthalein, and place it in a beaker A containing a solution of water and the base, sodium hydroxide (NaOH). Phenolphthalein is a base indicator that turns pink in basic solutions. 5. Fill bag B with 10 ml starch mixture and place it in a beaker B containing a solution of water and iodine. Iodine is a starch indicator that turns dark blue in the presence of starch. 6. Observe color change in the two bags contents and the surrounding solutions. 3

5 Osmosis of Decalcified Eggs Placed in Isotonic, Hypertonic, Hypotonic Solutions 4. The instructor will need three student volunteers for this experiment. 5. The initial weight of the eggs will be recorded to the nearest 0.1 gram and be placed in three different solutions: DI water, 25% sucrose, and 50% sucrose. 6. One hour later, the student volunteers will remove the eggs from the labeled beakers, blot them dry with a paper towel, and record the final weight to the nearest 0.1 gram. Effect of Molecular Weight on Diffusion 1. Create four holes on an agar plate and fill one with each of the following solutions, potassium permanganate (molecular weight = 158g/mole), methyl orange (molecular weight = 327g/mole), malachite green (molecular weight = 929g/mole, and methylene blue (molecular weight = 374g/mole) at the same time and allow them to diffuse. 2. Observe and measure the radius of the halos of color with a ruler for all demos. The halos show how far the substance has diffused through the agar from its original position. Biggest halo to smallest. Green (929)> Orange (327)> Blue (374)> Permanganate (158) *****WE MIGHT HAVE BAD POTASSIUM PERMANGANATE**** 4

6 Osmosis and Diffusion Name Osmosis a. Complete this table with data from your trial. Bags with 5% sucrose Initial Weight Final Weight Change in Weight % change = (finalw-initialw)/initialw*100 Descriptive Term (solution to bag) Bag #1 in 10.9g 11.8g 8.25% Hypotonic Distilled water Bag #2 in 5% sucrose 10.9g 10.9g 0% Isotonic Bag #3 in 25% sucrose 10.3g 7.2g % Hypertonic b. Did water move into, out of, or remain the same in the bags? Beaker #1 Into the bag Beaker #2 Remain Beaker #3 Out of the bag c. If a 10% sucrose cell model is placed into a 20% sucrose solution, which direction will water move? Water will flow into the 20% sucrose solution. d. Based on your observations of the cell models, is the dialysis tubing permeable to sucrose? Why? Yes, because glucose molecules will not move into the bag. Simple diffusion and Brownian motion a. Define diffusion. The movement of molecules from a higher concentration to a lower concentration. b. What causes the dye particles to vibrate, that is, why do dead molecules dance? What is name given to this driving force of diffusion? Brownian Motion 5

7 Diffusion of Substances through a Selectively Permeable Membrane a. Make a diagram of the initial set-ups for beakers 1 and 2, labeling what was present in each cell model and in each beaker solution. b. Describe the appearance of the bag and of the beaker solution in beaker 1 after time has passed. The bag containing phenolphthalein turned pink from inside. c. In beaker 1 which substances were able to pass through the selectively permeable membrane? Base Sodium Hydroxide (NaOH) d. Describe the appearance of the bag and of the beaker solution in beaker 2 after time has passed. The bag was grey-white but after 20 minutes the edges became black. e. In beaker 2 which substances were able to pass through the selectively permeable membrane? Iodine f. Why are some substances able to pass while others are restricted? Molecular weight of the molecules. g. Is the transport in these cases considered to be diffusion, osmosis or both? Why? 6

8 Osmosis of decalcified chicken eggs. a. Complete this table with data from your trial. Egg Initial Weight Final Weight Change in Weight % change = (finalw-initialw)/initialw*100 Descriptive Term In distilled water % Hypotonic In 25% sucrose % Isotonic In 50% sucrose % Hypertonic b. Did the water move into or out of the eggs in Beaker 1? The water moved into the egg. Beaker 2? The water moved out of the egg slightly. Beaker 3? The water moved out of the egg. c. Which egg gained the most weight? Why? The egg in distilled water gained the most weight because water is a hypotonic solution and the water followed the solutes inside the egg. Effect of Molecular Rate on Diffusion. a. Which would you predict would have a faster diffusion rate? Why? Potassium permanganate because it has a smaller molecular weight. b. Record the radius of the halo of The potassium permanganate: The methyl orange: The malachite green: The methylene blue: c. Which substance, the potassium permanganate or the methyl orange, had the largest halo? Is this what you predicted? Explain 7

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