You are doing a viable cell count for a culture of E. coli you are growing in a flask. The volume of media in the flask is 1L. You took an aliquot directly from the media and diluted it using trypan blue. Here are the numbers that you get: squares counted: 4 total cells counted: 988 total viable cells counted: 960 THESE ARE THE QUESTIONS: What is your cell viability? What is the total number of viable cells in your flask? Show all work. Given: Cell Viability = Viable Cells/Total Cells Cells/mL = Total Viable Cells/Coners Counted x 10,000 x Dilution factor

You are doing a viable cell count for a culture of E. coli you are growing in a flask. The volume of media in the flask is 1L. You took an aliquot directly from the media and diluted it using trypan blue. Here are the numbers that you get: squares counted: 4 total cells counted: 988 total viable cells counted: 960 THESE ARE THE QUESTIONS: What is your cell viability? What is the total number of viable cells in your flask? Show all work. Given: Cell Viability = Viable Cells/Total Cells Cells/mL = Total Viable Cells/Coners Counted x 10,000 x Dilution factor

Basic Clinical Laboratory Techniques 6E

6th Edition

ISBN:9781133893943

Author:ESTRIDGE

Publisher:ESTRIDGE

Chapter4: Basic Immunology And Immunohematology

Section4.3: Tests For Rheumatoid Factors

Problem 2.1CS

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Please note that these counts have all been done with Trypan Blue. Therefore, all cultures have been diluted two-fold (equal volume of cell suspension + equal volume of Trypan Blue). Take this into account when determining cell number. Any dilution listed in table is an additional dilution. Live and dead cells listed below represent the total number of cells in 5 large counting squares. *1 x 105 cells/ml cells were seeded into each well of the 24 well plate(s) on day 0 to begin the growth study. Plot this value at the zero time point on your graph.1These numbers represent the total number of cells counted in 5 large squares from three separate counts.
Why is the plate count method a determination of the number of viable cells?
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?
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. 輯 1 mm I IRE
After taking an aliquot of your cells and diluting them with Trypan Blue in a 1:1 dilution, you load the cell/Trypan mixture onto a hemocytometer to count the cells. You obtain the following counts: Quadrant Transparent cells Blue cells 1 2 3 4 LO 65 5 71 68 73 2 4 1 3 67 4) If your original cell culture (the one you took an aliquot from to count) has 13 mL of cell suspension, what is the total number of cells you have in culture? Provide your answer as the full number 2
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 .…
You are determining the concentration of your culture, and have counted the following on your hemocytometer as shown in Figure below: (note, blue cells have taken up Trypan blue & thus are dead. Red x's are cells that should not be counted because touching one of 2 sides of square). If your cells were diluted 1:2 before counting, what is the concentration of cells in your original culture?
You need to prepare medium for your culture cells. Your salt solution is supplied at a 10X concentration but needs to be 2X for use. You also need to add fetal bovine serum for a final concentration of 8%. What would you add of each for the correct final concentrations in 5 L of media?
To determine the number of cells in a pure culture of Klebsiella pneumoniae, you have performed a serial dilution using three tubes of sterile saline (9.9 ml each). A sample of 0.1 ml from the culture was added to tube 1. Similarly, 0.1 ml from tube 1 was used to inoculate tube 2, and tube 3 was inoculated using 0.1 ml from tube 2. A nutrient agar plate was then inoculated using 0.1 ml from tube 3 and incubated overnight. The next day, 92 colonies were observed on the plate. How many cfu/ml were in the original culture? Using a Petroff-Hauser counting chamber, the number of cells in the same culture was estimated to be 8.5 • 109 cells/ml. How can you explain these results?
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 Manton-Gaulin homogenizer is used for cell lysis. Which of the following are true? Increasing the number of passes can increase fine cell debris and make subsequent clarification more difficult. Cell growth conditions and phase of growth at which cells are processed can impact cell disruption. Decreasing temperature can increase protein release. Decreasing pressure reduces the number of passes required. Explain what is meant by the extent of cell disruption is 0.9.
You have several different media onto which you inoculated eight strains of yeast (A-H). The media include a rich medium, an unsupplemented minimal medium, and minimal media each supplemented with one vitamin. Of the yeast strains, one is a prototroph and seven are auxotrophs for a vitamin. After overnight incubation, the following results were observed (tan patches represent growth): D plate 1 (A) B DE F GH plate 5 plate 4 plate 6 Which plate contains an unsupplemented minimal medium? [Select] Which plate contains a rich medium? [Select] plate 2 Which strain is a prototroph? [Select] Strain E is an auxotroph for niacin. Which plate reveals this specific auxotrophy? [ Select] plate 3 plate 7 One strain is an auxotroph for both choline and pantothenic acid. Which one is this most likely to be? [Select]