Osmosis background Osmosis is the passive transport of water and occurs due to differences in the total amount of dissolved

molecules (solute) between two areas (Lluka 2010). Water will move, by osmosis, from areas of low solute concentration to areas of high solute concentration, until equilibrium is reached (Figure 1). It is not the concentration of specific molecules which determines osmotic movement, but rather the total concentration of all dissolved substances (the osmolarity). In living organisms, the direction and rate of osmotic movement has profound effects on bodily function (Good et al. 2008).

Figure 1: water moves by osmosis to equilibrate solute concentrations (Campbell et al. 2009).

In order to understand and predict the osmotic movement of water, it is essential to be able to accurately calculate the concentrations of different solutions. Concentration is typically expressed as molarity. Molar mass is the weight of 1 mole of a chemical compound; dissolving this mass in 1 litre of water will yield a 1 molar (M) solution. In order to make a 1 M calcium chloride (CaCl2) solution: 40.1 g mol-1 (molar mass of Ca) + (2 * 35.4 g mol-1) (molar mass Cl) = 110.9 g added to 1 l of water If a cell with an intracellular fluid (ICF) concentration of 150 mM is placed into a solution with an extracellular fluid (ECF) concentration of 150 mM, the ECF is isotonic to the ICF; the pressure on either side of the cell membrane is equal and the amount of water leaving and entering the cell will be equivalent. If the ECF has a lower concentration than the ICF (is hypotonic to the ICF) the amount of water entering the cell will be larger than that leaving the cell. If the ECF has a higher concentration than the ICF (is hypertonic to the ICF) the amount of water leaving the cell will be larger than that entering the cell.

Maintenance of constant body water content is essential to life: too little water can result in cellular dehydration and shrinking, and eventual death; too much water can lead to cell swelling, potentially breaking the cellular membrane, resulting in death. The introductory lectures have described the plasma membrane of cells. The plasma membrane acts as a barrier which effectively separates the ICF and ECF compartments. However, the membrane does permit the movement of certain substances between the ICF and ECF. The precise regulation of the membrane permeability underlies not only the osmotic movement of water, but also the physiological processes of cells. Linked to the concept of molarity is that of osmolarity. Osmolarity is a measure of the combined concentration of all solutes dissolved in a given volume of solution; the greater the concentration of dissolved ions, the greater the osmolarity. Osmolarity (in the units of Osm l-1) can be calculated in the following manner: Osmolarity (Osm l-1) = Concentration (M) * Number of ions solute dissociates into

Taking the example of a 1M calcium chloride solution again, the osmolarity would be calculated as follows: Osmolarity Osmolarity = = 1 * -1 3 Osm l 3 (1 Ca + 2 Cl)

Note: you will also need to refer to the guide on working scientifically in order to prepare for the osmosis practical class and pre-test. Experimental design: Now that you have a good understanding of the background material and how to work scientifically, you can design your own experiment ready for the practical class. You should have an outline of your experiment before you come to class, this will enable you to discuss materials and methods with your group members and tutor at the start of the class. Try to design a simple experiment that will measure an effect of both hypotonic and hypertonic ECF solutions on sheep red blood cells. You should base your hypotheses and materials and methods on what is achievable in the 3-hour time frame (it is better to have time spare at the end). The equipment below provides a range of different methods which can be used; you are not expected to use all of the equipment provided: Micropipettes accurately dispenses volumes of liquids. Whole sheep blood. NaCl (sodium chloride). 1-decimal place balance to accurately measure the mass of a compound. Distilled water. 100 ml volumetric flask accurately measures 100 ml of liquid to dissolve compounds.

Micro centrifuge separates whole blood into component parts for pipetting of ECF. Spectrophotometer measures the amount of colour in ECF. Haematocrit centrifuge - separates whole blood into component parts for Critocap reader. Critocap reader measures amount of space taken up by red blood cells in whole blood.

References: Campbell, N. A., J. B. Reece and N. Meyers (2009). Biology. Frenchs Forest, NSW, Pearson Education Asutralia. Good, J. P., A. Wells and N. Hazon (2008). "Measurement of blood volume in the elasmobranch fish Scyliorhinus canicula following acute and long-term salinity transfers." Journal of Fish Biology 73(6): 1301-1313. Lluka, L. (2010). Lecture 1.1: Osmosis. BIOL1040 Module 1 Lectures, The University of Queensland.