You want to subculture your cells from T25 flask to 96-well plates. You first collected your cells in a tube with 5ml of culture medium. Then carried out a trypan blue assay and counted your cells with a hematocytometer as shown in Figure below. Answer the questions according to the results: a. Calculate the concentration of the stock including the dead and living cells. Dont forget to show the units! b. Calculate the percentage (%) of the living cells in the stock. c. You want to seed 6000 living cells into each well of 96 well plates, then calculate the volume you should take from the stock for each well. 이 0 o 이.
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- You counted 4, 6, 12, 3 cells in each of the 4.outed squares of a hemacytomeyer. What are the cells per milliliter in that culture? If you resuspended your cell pellet 2.5 mL, what is the total cell count? How many uL do you need to add to a new culture if you want 4250 cells?In Figure 5-5,a. Why do A− and B− cells, by themselves, not formcolonies on the plating medium?b. What genetic event do the purple colonies in themiddle plate represent?Steven is a lab research assistant. He mixed 25 µL of cells with 50 µL of cell culture media in tube-A, then transferred 10 µL to the hemocytometer and started counting cells, then realized the cells were too crowded to count accurately, so Steven took an aliquot of 20 µL cells of cells from tube-A and mixed with 180 µL of cell culture media, transferred 10 µL onto the hemocytometer, and countered 500 cells from the squares on the four corners of the hemacytometer. What is the # of cells per mL? Please show all calculation steps.
- When analyzing your results after performing a serial dilution, you obtain an average of 40 colonies on your 1 x 106 plate. How many cells were in the original sample?At the station there is a 2 mL microcentrifuge tube containing sheep blood and 3 tubes containing the following solutions: 0 mM, 300 mM, 600 mM of sucrose. The tubes are randomly labelled A, B, C. 2 mL of each solution were transferred to a different microcentrifuge tube, 20 ul of blood were then added to each tube, and they were mixed well by inverting the tube for multiple times. They were observed and it was determined whether the solution in each tube was hypotonic (Yes or No) relative to the blood cells. In addition, the solutions were diluted with water to help you determine which solution is which sucrose concentration. What sucrose concentration corresponds with each tube? Tube Hypotonic (Yes/No) Hypotonic after 1:1 dilution with water (Yes/No) Most likely Sucrose concentration (mM) A No Yes B Yes Yes C No NoA.Why do you plate the cells from the viable count on LB agar without ampicillin? B.If you observe 100 colonies on your 1/100 plate, how many colonies do you expect if everything works perfectly on your 1/1000 plate?
- In a plate count, 1 mL of culture is spread on a plate and incubated for 24 hours. 250 colonies are counted. Answer the question below. there were _____cells in the 1 mL of culture that was spread on the plate. If the culture was diluted 1:100 prior to plating, how many cells were there per mL in the original culture? _________Following is the data and notice that it is a terrible idea to culture hMSCs longer than 10 days. You’re strongly Days # cells0 50001 75002 125003 125004 218005 287006 530007 1143008 1653009 19200010 19200011 11680012 8950013 8830014 78300 Part1 You are working for a start-up that is pursuing a clinical trial. The trial involves grafting hMSCs intopatients suffering from interveterbral disc disease using a degradable polymer scaffold. You are going to 3Dprint a porous cylindrical scaffold that is 2 cm in radius and 1 cm in height (matching the dimensions of adegenerated disc). Assume a porosity of 50%. You will fill available volume of the scaffold with hMSCs at adensity of 1 million cells per cm3. Based on the data above, what starting number of cells will you use andhow long will it take you to get enough cells for the trial? Part2The trial is a failure (patients did not report any reduction in back pain). Your team wants to try againusing 85% hMSCs and 15% nucleus pulposus cells .…Why is the plate count method a determination of the number of viable cells?
- In the ELISA, the pH the coating buffer was 9.6 whereas the pH of the sample buffer (PBS based) was 7.4 a) Why was the pH of the coating buffer so different?b) What is “coating buffer” for an ELISA? Does the buffer molecule usedmake sense based on the desired pH?In an ELISA procedure the samples are incubated and the ELISA plate iscovered with parafilm and placed in a humidified chamber to preventevaporation of the small liquid volumes in the wells.c) How would your results change if you did not incubate in a humidifiedchamber?You do a viral plaque assay using the six well plate shown below. The dilution scheme, what was infected on each well, and the results of the plaque assay are shown below. What is the viral titer of the original lysate? A. ~1.1 *10^6 PFU/mL B. ~1 *10^7 PFU/mL C. ~3 * 10^7 PFU/mL D. ~2 * 10^6 PFU/mL E. ~3.5 *10^6 PFU/mL F. ~1.1 * 10^5 PFU/mL G. ~3.5 *10^5 PFU/mLOf the 3 techniques that you used in this lab (direct fluorescent antibody, ouchterlony diffusion, ELISA) which do you think would work best for blood typing (A, B, O) and why?