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?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 .…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?
- The total number of cells in a culture is counted using the trypan blue exclusion assay and is found to be 6.8 x 106 cells/ml. Each well in a 6 well plate requires 2 x 105 cells. How should the solution be diluted so that 1ml can be added to each well?A pure bacterial culture of unknown concentration was diluted to determine the concentration of viable bacteria in the original culture. Serial dilutions were performed as 1. diagrammed below. Each dilution tube contained 400 ul of diluent and 100 ul was transferred into each tube. TSA plates were inoculated with 100 µul from the last three dilution tubes. a. What is the dilution between each tube shown in the diagram below? Express your answer as a ratio. b. What is the total dilution of tube number 5? Express your answer as a ratio. c. What is the concentration of viable bacteria in the original culture? Express your answer using scientific notation and the units CFU/ml. d. What is the concentration of viable bacteria in tube number 2? Express your answer using the units CFU/ml. e. If you inoculated a TSA plate with 250 µl from tube number 5, how many colonies would you expect to see after the plate was incubated? 1 2 3 5 423 80 13 Number of coloniesIn this experiment, different concentrations of sucrose are first prepared. Then, cylinders of potato (Solanum tuberosum) tuber are made using a cork borer, which are then trimmed to remove the skin and be of the same length. The cylinders are blot dried and weighed prior to being placed in each of the test tubes containing the sucrose solution. After 20 minutes, the cylinders were removed from the solution, blot dried, then weighed again. Study the results of the experiment shown in Table 1. Table 1. Changes in the weights of potato tuber tissue immersed in various concentrations of sucrose for 20 minutes Concentration (M) 0.0 0.2 0.4 0.6 0.8 1.0 Initial weight 3.76 3.15 3.29 3.82 3.40 3.52 Final weight 4.44 3.40 3.09 3.32 2.71 2.57 Change in weight (g) 0.68 0.25 -0.20 -0.50 -0.69 -0.95 % Change in weight 18.09 7.94 -6.08 -13.09 -20.29 -26.99 Calculate the water potential of the potato tuber cells. Express the answer in both bars and megapascal
- A pure bacterial culture of unknown concentration was diluted to determine the concentration of viable bacteria in the original culture. Serial dilutions were performed as 2. diagrammed below. A volume of 500 µl was transferred into each tube. TSA plates were inoculated with 100 µl from the last three dilution tubes. a. If the dilution between each tube is 102, what is the volume of diluent in each of the 5 dilution tubes? Provide the volume using ml as the units. b. What is the total dilution of tube number 4? Express the total dilution using scientific notation. c. What is the concentration of viable bacteria in the original culture? Express the concentration using scientific notation and CFU/ml as the units. d. If you inoculated a TSA plate with 1.0 ml from dilution tube 4, how many colonies would you expect to form on the plate after incubation? e. If the original culture had a volume of 50ml, what was the total number of viable bacteria in the 50 ml of the original culture? 1 2 3…A bacterial culture was grown for 12 hours. At 4-hour interval, the culture was sampled to determine the population of the culture, by transferring 25 ml of the suspension to 225 ml 0.90% NaCl. Three consecutive dilutions were further made by using I ml aliquot in 9 ml of 0.90% NaCl. One ml from each dilution was plated in each of duplicate plates. The following table shows the results of the plating method. 田 Sampling COUNTS 1st dilution 54; 61 2nd dilution 3rd dilution 4h dilution 1st 3; 7 0; 0 0,0 2nd TNTC TNCT 242: 233 28: 37 3rd TNTC TNTC INTC 249-246 * TNTC = Too numerous to count i. Illustrate the dilution series used and label the final dilution of each dilution. ii. Determine the bacterial count (CFU/ml) every 4 hours of incubation for 12 hours. Show all computations.Calculate the concentration of bacteriophage in the original culture from the following data. Be sure to include units. Show dilution factors for each test tube. Show the final dilution factor for test tube number 4. Show all your math. 0.1 ml 0.01 ml 0.001 ml 0.1 ml 1.0 ml plated 9.9 ml 9.99 ml 9.999 mi 9.9 m plaques Original Culture Test Test Test Test Tube #1 Tube #2 Tube #3 Tube #4 Dilution Factor: Concentration of Phage:
- Please count the cells in grids 1, 3, 7, and 9. Calculate the cell density (a) and cell viability (b) for this sample. (assume we started with 100 microliters of cells and added 100 microliters of trypan viability blue). Hemacytometer Grid 123 5 6 789 4 Enlarged view O O O 8When 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?Jon performed a pour plate technique to determine the concentration of viable cells in her mother culture for nata de coco production. Prior to all the samples, he poured an extra Petri dish without any sample with the molten agar. What is the purpose of doing this?