Name: Ranessa Dockery I.D #: 27100017 Course: Immunohematology Lab Instructor: Sala Randall Date: January 22, 2013 Lab #: 1 Title: Cell Washing Aim: To demonstrate quality assurance techniques in performing cell washing. Principle: Cell washing is a common procedure performed in the laboratory. This is a series of careful steps taken to wash red blood cells normally three times intermittently with centrifugation and decanting (Harmening 2012). The procedure serves to remove unbound immunoglobulins, Wharton’s jelly (from cord blood), hemolysed cells and also fibrin. The principle of the centrifuge entails centrifugal filtering of components due to centrifugal force applied and also centrifugal sedimentation …show more content…
The water was discarded. * Steps 3 to 6 were repeated twice. * The final pellet was re-suspended until a cherry red color (similar to 2-5% standards) was obtained. Results: It was observed that the more the sample was washed, the clearer the supernatant appeared and the smaller the pellet became. The final pellet was re-suspended until a cherry red color was observed. The colors and volume of the 2% and 5% standards were also observed and compared to that of the sample since the aim was to achieve a 2-5% suspension. The 2% standard was of a paler red than the 5%. The final suspension of the sample was observed to be between the two standards, however, closer to the color of the 5% standard after further comparison while the volume was equal to that of the 5% standard. Discussion: In cell washing 0.85% NaCl was used to wash the cell each time. This solution was made prior to the lab exercise, and not overnight or days ahead, to ensure that the concentration remained the same as required and no evaporation could take place to alter / increase the such, as this would have adverse effects on the cells. This concentration was chosen since it was isotonic to the concentration of the red blood cells. Since both solutions were isotonic no particle movement occurred and the cells were not altered in
Explain what happened to the blood cells at the various levels of concentration. Be sure to refer to the solutions as being hypotonic, hypertonic and isotonic.
In this lab experiment, half our group observed and measured osmosis using dialysis tubes that were represented as the semipermeable membrane. It is permeable to water and other small molecules but is impermeable to larger molecules such as the sucrose solution used in each of the four beakers and tubing. The other half of our group observed the tonicity of sheep blood to determine whether the blood was isotonic, hypotonic, or hypertonic. The 85 g/dL of NaCl solution was the ideal isotonic number in relation to the sheep blood cells as well as a reference to the other observations of the solutions.
Blood is a non-Newtonian fluid that contains many components. One such component is red blood cells. Due to the red blood cells having the tendency to clump together at low velocities, calcium chloride was added in order to cause a thrombus formation. The blood used in this experiment was sheep’s blood.
With all solutes set at a concentration of 5.00 mg/ml and the MWCO set at 20, filtration stopped at 60 minutes, and the projected completion was 100 minutes. The residue analysis indicated all solutes present in the dialysis membrane. The filtrate concentrations for all solutes was 0.00 mg/ml. With all solutes set at a concentration of 5.00 mg/m and the MWCO set at 50, the filtration completed in 40 minutes. The residue analysis indicated all solutes present in the dialysis membrane. The filtrate concentration for NaCl was 4.81 mg/ml, and 0.00 mg/ml for all remaining
The cystic fibrosis cell had an increasing mass as time went by because of osmosis. The normal cell had an almost constant mass. The normal cell played as a control group and the cystic fibrosis played as an experimental group in this experiment. Sodium and Chlorine were used for osmosis to determine how the cells differ when the amount of NaCl is changed. The independent variable used in this experiment was the amount of NaCl, and the dependent variable was the mass of the dialysis bag.
The same solution of 0.5 ml BSA was then added from test tube 1 to the test tube 2 after being properly mixed, and from test tube 2 the solution was being added to test tube 3, and so forth all the way up to test tube 5, with the same exact procedure. From the last tube, we then disposed the 0.5 ml solution. After above procedures, we now labeled another test tube “blank”; 0.5 ml blank distilled water was purred into the tube with the serial dilution of 1:10. We also had a tube C labeled “unknown” with the same 0.5 ml of solution. And after adding 5ml of Coomassie Blue to each tube (1-5) and to the blank, the result of absorbance was read at 595 nm.
The purpose of this experiment is to prepare Allura Red standard solutions, use a colorimeter to measure the absorbance value of each standard solution, find the relationship between absorbance and concentration of a solution and use the results of this experiment to determine the concentration of Allura Red
1.2 ml of the bacterial cultures are taken to pellet the cells and remove supernatant while
Blood is a non-newtonian fluid that contains many components. One such component is red blood cells. Due to the red blood cells having the tendency to clump together at low velocities, calcium chloride was added in order to cause a thrombus formation. The blood used in their experiment was sheep’s blood.
Each solution contained different concentrations as follows: 0.005 mg/mL, 0.010 mg/mL, 0.015 mg/mL, 0.020 mg/mL, and 0.025 mg/mL. Each solution needed to have a volume of 10 mL. Before adding the different concentrations of Coomassie Blue into their separate tubes, the formula C1V1= C2V2 was used in order to determine how much stock solution is needed for the five dilute solutions. Once that number was calculated, a pipette was used to add the amount of stock solution needed for each tube. We then subtracted the amount of stock solution from 10 mL to determine the amount of H2O needed. The calculated amount of H2O was then added to each tube of solution. After doing that, a spectrophotometer was used to determine each solution’s relative absorbance. However, before that, we first had to calibrate the spectrophotometer before determining each solution’s relative absorbance. In order to calibrate the spectrophotometer, a disposable culture tube filled with distilled water was used. We then changed the data rate to 100 and removed the tube with water. In order to determine the relative absorbance, the relative absorbance had to be at 595 nm. Also, during this experiment, an unknown dilution was given to us by the lab instructor. We determined the relative absorbance by using the spectrophotometer and then recorded the results. The procedures for this experiment can be found on page 8 of
The purpose of this lab is to see the effects of pasteurization while emphasizing the process for serial dilutions.
The purpose of this experiment was to determine the effects of tonicity on a cell membrane using red blood cells, potato strips and three unknown solutions (A, B, C). First three slides were prepared containing RBC’s and unknown solutions A, B and C. A control slide was prepared only using RBC’s. After observing each slide under the microscope it was determined that unknown solution A was hypertonic because the RBC appeared to have shrunk. The RBC in unknown solution B appeared to be swollen, therefor, the tonicity of unknown solution B was hypotonic. Unknown solution C showed no change to the RBC shape, it was suggested that unknown solution C was isotonic. To confirm the tonicity
The first solution used is distilled water which is a hypotonic solution. In this situation, water will diffuse into the red blood cells causing them to expand and be round (as shown in the above results) and sometimes they may rapture as shown in the picture below:
The conclusion that was made from this experiment was that as the streaking is completed the consistency as well as the concentration degrees allowing for a more accurate identification of the individual particles. The results lead me to this conclusion because as the streaks were completed the
When RBCs are suspended in an isotonic solution, nothing should be observed as there will be no net water movement between the RBCs and the solution. Thus, when 200 µL of blood was added to 10mL isotonic saline (0,154 M NaCl), voltage of 0V is recorded. When RBCs are suspended in a hypertonic solution, for instance, 0.4 M NaCl, RBCs shrunk due to decrease in cell volume as water diffused out of the cells by osmosis. The protein concentration within the cells become greater and more light is scattered and a negative