Effect of Solvent Exposure and Temperature on Beet Cell Membrane

The Effect of Solvent Exposure and Temperature on common beet Cell Membrane Integrity Introduction Cell membranes atomic number 18 resilient to proper carrelular function Cell membranes consist of a lipid bilayer made up of phospholipids Phospholipids are amphipathic molecules meaning they have a threefold spirit in that they show both hydrophilic (water-loving) and hydrophobic (water-fearing) properties The amphipathic nature of phospholipids bewilders them to spontaneously form bilayers in water based solutions o The inward and outer surfaces of the bilayers are hydrophilic and screwing interact with the aqueous environment, piece the core of the bilayer is hydrophobic and able to exclude water and other glacial molecules o Because of these properties, phospholipids bilayers are able to serve multiple functions including ? act as barriers to protect the jail cellular phoneular telephone from the outside environment ? house proteins and carbohydrates necessary for cell to cell and extracellular communication ? rganize and provide surface areas for metabolic reactions magical spell the phospholipids bilayer is a exceedingly organized system with multiple functions, it is also highly dynamic and can be functionally described using the swimming mosaic model (FMM) o FMM characterizes the phospholipids bilayer of the cell membrane as highly dynamic and fluid meaning that the position (location) of any given phospholipids in the membrane is not fixed but fluid thus, individual phospholipids can change positions are needed based on the current conditions. o Similarly proteins and carbohydrates are in general mobile in the membrane as well. This experiment was aimed at investigating the conditions that affect cell membrane single. o Because cell membranes are part of a living system they are sensitive to environmental conditions o Like most biological systems, cell membranes show a relatively condense range of function with keep an eye on to pH, temperature, and motion-picture show to organic resolvents The experimental model employed was red beets blushing(a) beet cells contain large, membrane bound central vacuoles that house a blusher, betacyanin o When cell and vacuole membranes are in tact, betacyanin in contained within the cell o If the cell and vacuole membranes are amputateed, betacyanin can leak in the touch solution turning it pink o Higher amounts of membrane trauma go forth lead to to a greater extent leakage of betacyanin hue from the cells. The more pigment present the high the absorbance reading. o Therefore, betacyanin release, as determined by bar absorbance, serves as a proxy for quantifying membrane cost. In this experiment, we tried the military unit of changing devil environmental conditions on cell membrane integrity.First, we tested the sum of temperature on cell membranes o To accomplish this we exposed beet cells to temperatures ranging from -5(C to 70(C o Because cell membra nes maintain integrity under a narrow range of conditions, we hypothesize that pic to higher temperatures (50-70(C) and extreme low temperatures (-5(C) entrust cause severe damage to the cell membranes which will result in higher amounts of betacyanin being released from the cells. We also tested the effect of exposing beet cells to two different solvents o Methanol and acetone are organic solvents equal of dissolving hydrophobic species such as phospholipids o Acetone is generally considered a stronger organic solvent compared with methanol o Thus, we hypothesize that exposure to acetone will cause more damage to beet cell membranes compared with exposure to methanol.Additionally, we hypothesize that higher concentrations of acetone will cause more damage than frown concentrations. Similarly, higher concentrations of methanol will also cause more damage. Results (the data numbers are for example only and meant to shine the general trend in the class data from individual grou ps will vary) Experiment 1 The effect of temperature on cell membrane integrity o We tested the effect of exposing beet cells to various temperatures ranging from -5(C to 70(C. (Question) o To do this, uniform pieces of beet were placed in empty test thermionic valves and so incubated at a specific temperature. (How the question was addressed) ? spare-time activity this water was added to the test tube and the cells were incubated for 20 minutes. ? After incubation, the water was transferred to a clean tube and absorbance at 425 nm was determined. o Absorbace readings were similar at temperatures of 5? C, 25? C, 40? C, 55? C, and 70? C (A425 0. 25-0. 30) ( mental image 1) (Experimental data) o Incubation at -5? C showed a dramatic affix in absorbance (A460 = 0. 95) (Figure 1) (Experimental data) o We shut down from this data that temperatures ranging from 5-70? C have only minor effects on cell membrane integrity. In contrast exposure of cells to a temperature under freezing c auses much more membrane damage. (Conclusion) Experiment 2 The effect of solvent on cell membrane integrity o We tested whether exposure to two different organic solvents, methanol and acetone, disrupted cell membrane integrity (Question) o Beet cells were incubated in 10%, 25%, and 50% methanol or 10%, 25%, and 50% acetone. Additionally, additionally cells were also incubated in saline solution, a solvent that mimics physiological conditions. (How the question was addressed) ? by-line exposure to solvent, membrane damage was determined by measuring absorbance. o Incubation in 10% or 25% methanol yielded absorbance readings of 0. 2 and 0. 25 respectively however, incubation in 50% methanol showed an increase in absorbance to 0. 5. Figure 2) (Experimental data) o Incubation in 10% or 25% acetone produced higher absorbance readings compared to the same concentrations of methanol (0. 4 and 0. 42 respectively). Similar to 50% methanol, 50% acetone yielded the highest absorbance readin g of 0. 8 (Figure 2). (Experimental data) o Taken together, these data show that acetone causes more membrane damage than methanol and that concentrations of 50% solvent are more damaging than lower concentrations. (Conclusion) Overall Conclusion We hypothesized that temperatures outside of the normal physiological range would disrupt membrane integrity causing betacyanin to leak out of the cells. Our data shows that our scheme was partly correct, low temperature, -5? C, inflicts the most damage to cell membranes while higher temperatures, ranging from 55-70? C induce little damage when compared with physiological temperatures. We also hypothesized that exposure to high concentration organic solvents would disrupt membrane integrity and that acetone, being a stronger solvent, would show more damage than methanol. o In this skid our hypothesis was support. Acetone showed more damage as measured by betacyanin release than methanol. Additionally, the highest concentration of each s olvent showed the most damage. Data pic Figure 1 The effect of temperature on cell membrane integrity.Cells were exposed to temperatures ranging from -5C to 70C. Following incubation at the indicated temperature, water was added and the cells were allowed to incubate at room temperature for 20 minutes. Damage to cell membrane integrity was assessed by quantifying leakage of the pigment betacyanin via measuring light absorbance at 460 nm. pic Figure 2 The effect of solvent on cell membrane integrity. Cells were incubated in the presence of methanol, acetone, or saline at the indicated concentrations. Following incubation, damage to cell membrane integrity was assessed by quantifying leakage of the pigment betacyanin via measuring light absorbance at 460 nm.