Introduction Sold commercially under the tradename Sevin, carbaryl (C12H11NO2) is an insecticide that is commonly applied to corn, soybean, and cotton (EPA, 2000). It is a cholinesterase inhibitor that kills insects by preventing the breakdown of acetylcholine, a neurotransmitter. Neurons are stimulated when they release acetylcholine and transmit a message to the next neuron (Waymire, n.d.). However, in order for a neuron to return to its initial, unstimulated state, the acetylcholine in its receptor must be broken down into acetyl and choline. Cholinesterase is the chief enzyme that is responsible for the reduction of acetylcholine (Waymire, n.d.). Because cholinesterase is inhibited upon the application of carbaryl, acetylcholine is prevented
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Four tubes were treated with 5 µL of commercial carbaryl at concentrations of 0 M (control), 1.19 mM, 11.9 mM, and 119 mM. The other four tubes were treated with 5 µL of pure carbaryl at concentrations of 0 M (control), 5 µM, 50 µM, and 500 µM. The treated microcentrifuge tubes were then allowed to incubate in a refrigerator at 4 °C for one hour. After the one-hour period elapsed, the treated hemocytes were dyed with 5 µL of trypan blue and viewed under the microscope using a hemocytometer. Under the microscope, the dead hemocytes appeared dyed blue, whereas the alive ones were translucent. A cell count was performed in multiple locations of each quadrant in the hemocytometer calculate the percent of cells that were dead after one-hour treatment with carbaryl. There were a total of sixteen trials of cell counts for each treatment.
Cell Viability Assay on …show more content…
Each well was then rinsed out with 500 µL of distilled water before adding 300 µL of Hema-3 fixative, which fixed the cells to the bottom of the plates. After allowing the wells to sit for a minute, the fixative solution was removed and 300 µL of Hema-3 Solution II, a type of cell dye, were added to each well of the plate. After another minute, the dye was removed and the plate was rinsed under a trickle of water until no more visible dye was left over. The plate was then examined under a microscope at 4 times magnification, and in each well, the number of cells that stuck to the bottom of the plate were counted for a total of 10 trials per
Four tubes were treated with 5 µL of commercial carbaryl at concentrations of 0 M (control), 1.19 mM, 11.9 mM, and 119 mM. The other four tubes were treated with 5 µL of pure carbaryl at concentrations of 0 M (control), 5 µM, 50 µM, and 500 µM. The treated microcentrifuge tubes were then allowed to incubate in a refrigerator at 4 °C for one hour. After the one-hour period elapsed, the treated hemocytes were dyed with 5 µL of trypan blue and viewed under the microscope using a hemocytometer. Under the microscope, the dead hemocytes appeared dyed blue, whereas the alive ones were translucent. A cell count was performed in multiple locations of each quadrant in the hemocytometer calculate the percent of cells that were dead after one-hour treatment with carbaryl. There were a total of sixteen trials of cell counts for each treatment.
Cell Viability Assay on …show more content…
Each well was then rinsed out with 500 µL of distilled water before adding 300 µL of Hema-3 fixative, which fixed the cells to the bottom of the plates. After allowing the wells to sit for a minute, the fixative solution was removed and 300 µL of Hema-3 Solution II, a type of cell dye, were added to each well of the plate. After another minute, the dye was removed and the plate was rinsed under a trickle of water until no more visible dye was left over. The plate was then examined under a microscope at 4 times magnification, and in each well, the number of cells that stuck to the bottom of the plate were counted for a total of 10 trials per