Effect of Solvent Exposure and Temperature on Beet Cell Membrane

The Effect of Solvent Exposure and Temperature on Beet Cell tissue layer Integrity Introduction Cell tissue layers atomic number 18 vital to proper stallular function Cell tissue layers consist of a lipid bilayer made up of phospholipids Phospholipids argon amphipathic molecules meaning they have a dual nature in that they demonstrate both hydrophilic (water-loving) and aquaphobic (water-fearing) properties The amphipathic nature of phospholipids causes them to spontaneously stock bilayers in water based solutions o The inner and outer surfaces of the bilayers argon hydrophilic and bottom interact with the aqueous environment, while the core of the bilayer is hydrophobic and able to exclude water and other polar molecules o Because of these properties, phospholipids bilayers argon able to serve multiple functions including ? act as barriers to protect the mobile phone from the outside environment ? house proteins and carbohydrates necessary for booth to cadre and extra c ellphoneular communication ? rganize and provide surface areas for metabolic reactions While the phospholipids bilayer is a extremely organized system with multiple functions, it is also highly dynamic and can be functionally described using the politic mosaic baffle (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 generally mobile in the membrane as well. This experiment was aimed at investigating the conditions that affect cell membrane uprightness. 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 narrow range of function with respect to pH, temperature, and exposure to thoroughgoing solvents The experimental model employed was red common beets Red beet cells check off large, membrane bound central vacuoles that house a pigment, betacyanin o When cell and vacuole membranes are in tact, betacyanin in contained within the cell o If the cell and vacuole membranes are disrupted, betacyanin can leak in the surrounding solution bout it pink o Higher amounts of membrane damage volition work to more leakage of betacyanin pigment from the cells. The more pigment present the higher(prenominal) the absorbance reading. o Therefore, betacyanin release, as determined by measuring absorbance, serves as a proxy for quantifying membrane damage. In this experiment, we tried the set up of changing 2 environmental conditions on cell membrane righteousness.First, we tested the effect 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 membranes maintain integrity under a narrow range of conditions, we opine that exposure to higher temperatures (50-70(C) and extreme pitiful temperatures (-5(C) will cause unspeakable damage to the cell membranes which will result in higher amounts of betacyanin existence released from the cells. We also tested the effect of exposing beet cells to twain different solvents o Methanol and acetone are total solvents capable of dissolving hydrophobic species such as phospholipids o acetone is generally considered a stronger organic solvent compared with wood spirit o Thus, we say that exposure to acetone will cause more damage to beet cell membranes compared with exposure to methanol.Additionally, we hypothesize that higher minginesss of acetone will cause more damage than humble immersions. Similarly, higher concentrations of methanol will also cause more damage. Results (the info numbers are for example only and meant to reflect the general swerve in the class selective information from individual groups will v ary) 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 tubes then incubated at a specific temperature. (How the question was addressed) ? pursuit 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) ( auspicate 1) (Experimental data) o Incubation at -5? C showed a dramatic increase in absorbance (A460 = 0. 95) (Figure 1) (Experimental data) o We conclude 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 below freezing causes overmuch more membrane damage. ( s hutdown) 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, a solvent that mimics physiological conditions. (How the question was addressed) ? Following 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 reading of 0. 8 (Figure 2). (Experimental data) o T aken 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 hypothesis was partly correct, low temperature, -5? C, inflicts the most damage to cell membranes while higher temperatures, ranging from 55-70? C brace 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 case our hypothesis was support. Acetone showed more damage as measured by betacyanin release than methanol. Additionally, the highest concentration of each solvent showed the most damage. Data pic Fig ure 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 set out 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.