Lab Note #4 Second experiment Since the previous result was not clear and cells did not get reduced as much as I thought, I decided to the second experiment by putting an increased concentration of PLA2 and 5FU on colorectal cells. Method: 1. I took plates with cells(WIDR and HCT15) out of a culture medium. 2. I got rid of liquid media using a suction machine. 3. I put 500ul of TE buffer so that cells on the bottom of the plates float. After I put them on the centrifuge I waited for 5 minutes. (If the cells are on TE buffer too long, cells can die.) 4. After 5 minutes, I slightly shook the plate with TE buffer and cells so that the cells are detached from the bottom of the plates. 5. I put them on two 1.5ml tubes. 6. I put the tubes on a centrifuge that spin in a speed of 7500 rpm for 2 minutes. Then I can saw cells on the bottom of the tube separated from TE buffer. 7. I sucked TE buffer carefully so that the cells don't get sucked up together. 8. I put 200ul of RPMI and d
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Lab Note #3 The result Experiment date: 2020. 8. 31 Material: Microscope, cell counting machine, the result of the first experiment 1. Observing the result of the last experiment Through a microscope, I observed colorectal cells that were treated with PLA2 and 5-FU. Then I took pictures of them through a computer connected to the microscope. Though I expected to observe the reduced cells, I was surprised to see the cells that reduced only in a small amount. So I decided to use cell counting machine to get the exact result. 2. Counting cells Dead cells are stained red while alive cells are stained green. The machine then count the number of green cells and red cells. First, I put small amount of buffer solution with chemical (PLA2 or 5FU) and cells on the small plate. (Pipetting should be done in order to make sure that cells aren't compacted together.) Then, I put the plate on the machine which starts counting. After I check the result, I saved it on the USB. Though the proce