Effect of Solvent Exposure and Temperature on Beet Cell Membrane

The Effect of Solvent pictorial matter and Temperature on Beet Cell tissue layer Integrity Introduction Cell membranes atomic number 18 vital to proper jail prison jail cellular matter Cell membranes consist of a lipoid bilayer made up of phospholipids Phospholipids atomic number 18 amphipathic molecules means they have a dual reputation in that they represent both deliquescent (water-loving) and hydrophobic (water-fearing) properties The amphipathic nature of phospholipids causes them to spontaneously fig bilayers in water based solutions o The inner and outer surfaces of the bilayers are hydrophilic and can interact with the aqueous environment, date the core of the bilayer is hydrophobic and able to eliminate water and other polar molecules o Because of these properties, phospholipids bilayers are able to serve threefold designs including ? act as barriers to protect the cell from the outside environment ? house proteins and carbohydrates needful for cell to cell and extracellular communion ? rganize and provide surface areas for metabolic reactions maculation the phospholipids bilayer is a upliftedly organized musical arrangement with multiple functions, it is as well as highly slashing and can be functionally draw using the fluid mosaic precedent (FMM) o FMM characterizes the phospholipids bilayer of the cell membrane as highly dynamic and fluid marrow that the position (location) of any given phospholipids in the membrane is not fixed nevertheless fluid thus, individual phospholipids can tack 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 study the conditions that affect cell membrane fair play. o Because cell membranes are part of a living system they are tenuous to environmental conditions o Like al approximately biological systems, cell membranes bespeak a relatively narrow range of fun ction with respect to pH, temperature, and pic to organic resultants The experimental model employed was red beets ablaze(p) beet cells contain large, membrane bound central vacuoles that house a key, 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 turning it go o Higher amounts of membrane harm forget lead to to a greater extent than effluence of betacyanin pigment from the cells. The more(prenominal) pigment look the higher(prenominal) the absorbance reading. o Therefore, betacyanin release, as intractable by touchstone absorbance, serves as a proxy for quantifying membrane ill-use. In this experiment, we well-tried the onus of changing two environmental conditions on cell membrane uprightness.First, we well-tried the effect of temperature on cell membranes o To accomplish this we exposed beet cells to temperature s ranging from -5(C to 70(C o Because cell membranes maintain rightfulness to a outseter place a narrow range of conditions, we say that photo to higher temperatures (50-70(C) and extreme low temperatures (-5(C) will cause severe toll to the cell membranes which will result in higher amounts of betacyanin being released from the cells. We also time-tested the effect of exposing beet cells to two assorted resolving powers o Methanol and acetone are organic resolutions capable of dissolving hydrophobic species such as phospholipids o dimethyl ketone is generally considered a stronger organic solvent compared with methyl alcohol o Thus, we hypothesize that delineation to acetone will cause more vilify to beet cell membranes compared with image to methyl alcohol.Additionally, we hypothesize that higher concentrations of acetone will cause more damage than lower berth concentrations. Similarly, higher concentrations of methanol will also cause more damage. Results (the s elective information total are for example only and meant to beam the general trend in the course of instruction data from individual groups will vary) experimentation 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 move in empty test tubings consequently incubated at a specific temperature. (How the research was addressed) ? pursual this water was added to the test tube and the cells were incubated for 20 minutes. ? After brooding, 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) ( fig 1) (Experimental data) o brooding at -5? C showed a dramatic add-on in absorbance (A460 = 0. 95) (Figure 1) (Experimental data) o We conclude from this data that temperatures ranging from 5-70? C have only peasant effects on cell membrane integrity. In contrast exposure of cells to a temperature below freezing causes 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, a solvent that mimics physiologic 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 enlarge 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 metha nol (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 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 hypothesis 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 case our hypothesis was support. Acetone showe d more damage as measured by betacyanin release than methanol. Additionally, the highest concentration of each solvent 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 means 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.