The marvelous laboratory manner to study bacterial infection by cytotoxicity assay
Bacteria are one of the most diverse living organisms, which enable adapted to a great variety of environments including the human body(1). As we know, the Scientists are classified the bacteria to two classes: non- pathogenic bacteria or commensal organisms and pathogenic bacteria. Researchers are predominantly shed light on pathogenic bacteria due to these bacteria use a number of different virulence mechanisms that allow them to overcome the many different defensive lines of immune system in the host. Additionally, the bacterial virulence is a multifactorial process that requires the use of a variety of components, many of which are coordinately regulated …show more content…
It should remind that each of bacteria has specific types of toxins such as Pseudomonas secreted a different type of toxins including the Type III secreted cytotoxins, ExoS, ExoT, and ExoU, which have ability to cause the cell death. However, the Streptococcus pyogenes outputs a different type of toxin known hemolysins, which a play significant role for destroy the red blood cells. In this experiment, we will measure the effect of bacterial cytotoxins on red blood cells by spectrophotometer, which measures the amount of hemoglobin release for RBCs. For measure the amount of hemoglobin released from red blood cells in this lab, we should flow three principle ways: prepper RBCs, prepper the P.aerginosa, and finally prepare the co-culture for RBc with bacteria. The first stage was prepared culture RBD with bacteria at MOI=100:1, then, we transferred about 1 ml of sheep RBC’s to microfuge tube and spin 5’ at 6000rpm and remove about 500 µl supernatant from microfuge tube and throw it. Subsequently, we added about 500 µl of PBS on suspend RBCs that remain in microfuge tube. After that, we conveyed 1 µl of the suspension to 999 µl of PBS in another microfuge tube and mix …show more content…
We used the Staphylococcus epidermidis as negative control and that meaning we don expected to any effect. The huge level of hemoglobin released was from SDS and RBC and PBS about 0.368 due to the SDS or Sodium Dodecyl Sulfate worked as a detergent, which have ability to denture the protein structure, and destroy the cell wall, and disrupt the nucleic acid. Upon this result, we access to estimate the ability of Pseudomonas aeruginosa, Streptococcus pyogenes, and Staphylococcus epidermidis to cause eukaryotic cell death destroy the red blood cells and we achieve our
Bacteria are one of the most diverse living organisms, which enable adapted to a great variety of environments including the human body(1). As we know, the Scientists are classified the bacteria to two classes: non- pathogenic bacteria or commensal organisms and pathogenic bacteria. Researchers are predominantly shed light on pathogenic bacteria due to these bacteria use a number of different virulence mechanisms that allow them to overcome the many different defensive lines of immune system in the host. Additionally, the bacterial virulence is a multifactorial process that requires the use of a variety of components, many of which are coordinately regulated …show more content…
It should remind that each of bacteria has specific types of toxins such as Pseudomonas secreted a different type of toxins including the Type III secreted cytotoxins, ExoS, ExoT, and ExoU, which have ability to cause the cell death. However, the Streptococcus pyogenes outputs a different type of toxin known hemolysins, which a play significant role for destroy the red blood cells. In this experiment, we will measure the effect of bacterial cytotoxins on red blood cells by spectrophotometer, which measures the amount of hemoglobin release for RBCs. For measure the amount of hemoglobin released from red blood cells in this lab, we should flow three principle ways: prepper RBCs, prepper the P.aerginosa, and finally prepare the co-culture for RBc with bacteria. The first stage was prepared culture RBD with bacteria at MOI=100:1, then, we transferred about 1 ml of sheep RBC’s to microfuge tube and spin 5’ at 6000rpm and remove about 500 µl supernatant from microfuge tube and throw it. Subsequently, we added about 500 µl of PBS on suspend RBCs that remain in microfuge tube. After that, we conveyed 1 µl of the suspension to 999 µl of PBS in another microfuge tube and mix …show more content…
We used the Staphylococcus epidermidis as negative control and that meaning we don expected to any effect. The huge level of hemoglobin released was from SDS and RBC and PBS about 0.368 due to the SDS or Sodium Dodecyl Sulfate worked as a detergent, which have ability to denture the protein structure, and destroy the cell wall, and disrupt the nucleic acid. Upon this result, we access to estimate the ability of Pseudomonas aeruginosa, Streptococcus pyogenes, and Staphylococcus epidermidis to cause eukaryotic cell death destroy the red blood cells and we achieve our