Canine Endothelial Culture: A Case Study

In Exercise 1, diaminofluorene is used to determine the hemoglobin concentration in the daphnids. A higher hemoglobin concentration is indicated by a darker blue color. A spectrophotometer was used to determine the absorbance at 610nm. When measuring the absorbance levels a blank is necessary to have a zero reference, the blank is the “starting point” for the measurements of the sample (re-word). The blank consists of 10µL of diaminofluorene(DAF), 50µL of hydrogen peroxide, and 0.5mL of PBS. The PBS acts as a buffer in this experiment. The cuvette with the sample of daphnids consisted of 10µL of DAF, 50µL of hydrogen peroxide, and 0.5mL of the sample of Daphnia. In Exercise 2, the Pasteur pipette was used to obtain the sample of Daphnia. The depression slide used in this experiment isolated the daphnid, cotton was used to keep the daphnid still while the heartbeat was counted. The ocular micrometer on the microscope allows the tail spine length to be measured accurately, as well as using an ocular magnification table.

5.  What reaction would you expect when performing a negative control in the catalase assay?

The preparation for the experiment started by gathering the solutions of enzyme Peroxidase, substrate hydrogen peroxide, the indicator guaiacol and distilled water. Two small spectrometer tubes and three large test tubes with numbered labels. In addition, one test tube rack, one pipet pump and a box of kimwipes were also gathered. Before the experiment, the spectrometer must be set up to use by flipping the power switch to on. Following, the machine was warmed up for 10 minutes and the filter lever was moved to the left. In addition, I set the wavelength to 500 nm with the wavelength control knob. Before the experiment, I had to create the blank solution by pipetting 0.1 ml of guaiacol, 1.0 ml of turnip extract and 8.9 ml water into tube #1. Following the creation of the blank, a control 2% solution was created.

10.State a commercial use for biochemical testing as performed in this online laboratory. (5 points)

1. We measured 2 mL of diluted hydrogen peroxide (the substrate), 1 mL of guaiacol (the product indicator), and 1 mL of neutral buffer (pH 7) with a syringe and disposed it into tubes 1, 2 , 4, 9, 11, and 12.

A 24-well plate was used to perform this experiment. For the start of the experiment, the 24-well plate had 12 wells that contained our HepG2 cells. A cell suspension was created, that contained 0.5-1.0x10⁶ cells/ml containing 10% fetal bovine serum. 500µl of the cell suspension was then added to each well in use. The well plate contained both the HepG2 cells, and cell suspension was then incubated in a cell culture incubator for 24 hours; this allowed for a monolayer to be formed in the wells. After the incubation period elapsed, the well plate was removed and placed inside a tissue culture hood. We then carefully removed all the media from the wells with the use of a P1000 pipette; making sure not to touch the bottom of the well plate, and

After the substrate solution was added, five drops of the enzyme were quickly placed in tubes 3, 4 and 5. There were no drops of enzyme added in tubes 1 and 2 and in tube 6 ten drops were added. Once the enzyme solution has been added the tubes were then left to incubate for ten minutes and after five drops of DNSA solution were added to tubes 1 to 6. The tubes were then placed in a hot block at 80-90oC for five minutes. They were then taken out after the five minute period and using a 5 ml pipette, 5 ml of distilled water were added to the 6 tubes and mixed by inversion. Once everything was complete the 6 tubes were then taken to the Milton Roy Company Spectronic 21 and the absorbance of each tube was tested.

Colorimetric assay is a process of determining a concentration of a solution based on absorbance of light. The purpose of this lab is to determine if the Bradford assay is an accurate way to determine an unknown concentration of two samples of protein. The Bradford assay is done by measuring wavelength of light passing through a cuvette filled with Bradford dye and concentrations of PBS and proteins. After the cuvettes are mixed they are placed into a spectrophotometer to measure wavelength. The wavelength given will be used to plot a standard curve based on concentration (x-axis) and wavelength (y-axis). The standard curve is then used to measure an educated guess on the concentrations of unknown protein concentrations. We hypothesized that if we use the Bradford assay and colorimetric spectrophotometry we can determine an accurate concentration of two unknown concentrations of proteins. The results of this lab failed to reject our hypothesis based on accurate measurements of protein concentrations. The standard curves are drawn with a linear increasing slope. The Bradford assay is an accurate way to demine the concentration of an unknown concentration.

Test tubes 3A and 3B were placed in a water bath set to 70૦C. Lastly, test tubes 4A and 4B were set in a 250 ml beaker after water ⅔ of way had been boiling on a hot plate. To speed up the boiling process, pellets of boiling starters were added to the water-filled beaker. After each of the tubes were exposed to their proper element, a 5 minute timer was set. Once time was up, the tubes in Group B, which contained the hydrogen peroxide, was added to the test tubes in Group A, which contained the liver tissue sample and water. To clarify, test tube 1B was added to test tube 1A, test tube 2B was added to test tube 2A, test tube 3B was added to test tube 3A, and test tube 4B was added to test

So in order to see the effect of lead ions into the enzyme rate of

After patients have gone through heart disease, like any other injury, therapeutic steps must be taken. Therapy and rehabilitation of damaged hearts has been improved through the utilization of stem cells. Therapy includes strengthening of the damaged area as to prevent the incident from reoccurring and getting the patient back to their normal life. Scientists, again, used rats that had had induced myocardial infarctions. This

After the dogs had been anesthetized and given the injection of the endotoxin, blood was drawn. The blood was collected by use of a catheter in the femoral artery. However, for each sample, a new catheter was used. Between each collection of the samples, the artery had to be clamped using a Pott’s Clamp. The blood collected was then stored in a silicon tube. Nine units of whole blood were anticoagulated with one volume of 3.2% sodium citrate. The plasma was then separated by centrifugation for 30 minutes and stored in silicon test tubes at negative twenty

(This experiment is aimed to use 10 Artemia, however, 7 to 13 Artemias are able to achieve the goal in this experiment.). Then seal the cuvette under water and make sure no air bubble is inside the cuvette. After that, put the cuvette into the temperature controlled water bath for ten minutes. After 10 minutes, take the cuvette to the oxygen meter to measure the oxygen concentration by holding the end of the end of the fibre-optic cable squarely on to the senor spot from the outside of the cuvette until the concentration has been shown on screen and record it down. Then return the Artemia to the same incubation bath and repeat this procedure every 5 minutes and measure it for 4 to 5 times. After the process above, we have to find out the total length of the Artemias in the cuvette. To find the total length of the Artemia, use the pipette to move the Artemia out of the cuvette and settle them into a watch glass and measure the length of the Artemias by ruler. At last but not least, put those high energy intake Artemia back into the sink and repeat the experiment instead of those low intake Artemia. On the other hand, to find the difference of activity of the high and low energy intake Artemia, those Artemia will be tracked by the software named, the Tracker and the Tracker are able to determine the velocity of the Artemia under different treatment for 5 replicates.

Endothelial cells are those that are found in endothelium tissue. Endothelial tissue typically lines the surface of various blood and lymphatic vessels. The endothelial cells are in direct contact with the blood and lymphatic cells. More specifically, endothelial cells that come in contact with blood are called vascular endothelial cells. Sickle Cell Anemia is a blood disorder that results in an abnormality in the hemoglobin found in the red blood cells. Endothelial cells are involved with many functions of the cardiovascular system. High amounts of circulation endothelial cells have been known to be found in patients with Sickle Cell Anemia. The article examines the origin, surface phenotype, and function of circulating endothelial cells in patients with the disease.

On January, 2008, Doris Taylor, director, Center for Cardiovascular Repair, University of Minnesota used the core procedure of decellularisation to let hearts from animal cadavers beat by infusing live cells. The research