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Basic Clinical Laboratory Techniques 6E

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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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:

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1mL Stock #1 1mL 9mL #2 9mL #3 1mL A 9mL #4 0.1mL 1mL O O B 9mL #5 1mL 1mL 9mL #6 O Using the picture serial dilution scheme and the following information (plate A has 276 colonies, plate B has 298, plate C has 2, and plate D has 30), calculate the average number of colony forming units per mL in the stock tube. Make sure to only use countable plates. Round your answer to the nearest one. Write only the number with any needed commas or decimals. Do not include units. 0.1mL 1mk D O
A laboratory technician performed a series of 10 fold serial dilutions shown below. he plated 0.1 mL from each dilution tube onto NA and got the follow numbers per plate dilution 1:10 1:10 1:100 1:1000 1:10000 1:100000 colony count >1000 >1000 808 303 38 3 calculate the concentration of bacteria in the undiluted stock (in CFU/mL)
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 are given a bacterial culture which has a concentration of approximately 5.0 x 10^8 cells/mL. List a series of dilutions and platings that you could carry out in order to determine the exact concentration of the culture. Note that you must plate four plates from a minimum of two dilution tubes. The volumes plated should be in the range of 0.1 mL – 1.0 mL. Duplicate volumes may not be plated from any one dilution tube. Each plating should aim for a count between 30 and 300 CFUs. You can select any value from 30-300 for CFU and any volume from 0.1-1.0 to find out dilution scheme
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…
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 No
The number of bacterial cells in a culture broth is to be determined by a culture technique. Serial dilutions were performed and a 0.1 mL aliquot from each dilution was spread onto Plate Count Agar (PCA). The number of bacterial colony forming units (CFU) after overnight incubation are shown as listed in the table below. What is the number of colony forming units per mL of the culture broth? Choose only the most appropriate plate for your calculation. Give your answer as the number only (do not add text for the units). You may use scientific notation with the format 1.12e+6 (that is, 1.12 x 106 cfu/mL). (Note: Canvas will then display your answer a whole number.) Plate 1 10 Plate 2 10 Plate 3 10 dilution dilution dilution Plate 4 10 dilution Plate 5 107 dilution Plate 6 10 dilution -6 *Too many to count Number of colony forming units (CFU) TMTC* TMTC* 840 28 19 1
The number of bacterial cells in a culture broth is to be determined by a culture technique. Serial dilutions were performed and a 0.1 mL aliquot from each dilution was spread onto Plate Count Agar (PCA). The number of bacterial colony forming units (CFU) after overnight incubation are shown as listed in the table below. What is the number of colony forming units per mL of the culture broth? Choose only the most appropriate plate for your calculation. Give your answer as the number only (do not add text for the units). You may use scientific notation with the format 1.12e+6 (that is, 1.12 x 106 cfu/mL). (Note: Canvas will then display your answer a whole number.) Plate Dilution Plate 1 10 dilution Plate 2 10 dilution Plate 3 107 dilution Plate 4 10 dilution Plate 5 107 dilution Plate 6 100 dilution *Too many to count Number of colony forming units (CFU) TMTC* 382 83 10 2 0
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.
The following are errors that people commonly make when they perform serial dilutions. Indicate whether you think that the number of cfu/ ml calculated would be too high or too low if you make this mistake. You intend to add 0.9 ml of diluent to each tube and 0.1 ml of culture. Instead, you add 0.5 ml of diluent to each tube and 0.1 ml of culture to the first tube. Then, you make a serial dilution of 0.1 ml into and from each tube as described. You prepare 0.9 ml of diluents in each tube. You add 0.1 ml of culture (from the overnight culture provided) to every tube. You add 0.9 ml of diluent to each tube. You add 0.1 ml of culture to the first tube and mix. You get distracted, and transfer 0.1 ml to the third tube instead of the second. You perform the rest of the series as described.
The number of bacterial cells in a culture broth is to be determined by a culture technique. Serial dilutions were performed and a 1 mL aliquot from each dilution was mixed with warm molten agar and poured into a Petri dish. The numbers of bacterial colony forming units (CFU) after overnight incubation are shown in the table below. What is the number of colony forming units per mL of the culture broth? Choose only the most appropriate plate for your calculation. Give your answer as the number only (do not add text for the units). You may use scientific notation with the format 1.12e+6 (that is, 1.12 x 106 cfu/mL). (Note: Canvas will then display your answer a whole number.) Plate Dilution Plate 1 10 dilution Plate 2 10 dilution -5 Plate 3 10 dilution Plate 4 10 dilution Plate 5 107 dilution -8 Plate 6 100 dilution *Too many to count Number of colony forming units (CFU) TMTC* TMTC* TMTC* 867 154 18
Ten grams of hamburger were added to 90 mL of sterile buffer. This was mixed well in a blender. One-tenth of amL of this slurry was added to 9.9 mL of sterile buffer. After thorough mixing, this suspension was further dilutedby successive 1/100 and 1/10 dilutions. One-tenth of a mL of this final solution was plated onto Plate Count agar.After incubation 145 colonies were present. How many colony-forming units were present in the total 10 gramsample of hamburger?

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