Detecting Circulating Tumor Cells using Flow Cytometry Essay

Detecting Circulating Tumor Cells using Flow Cytometry - Essay Example The research field was on Flow Cytometry. It aimed to  establish  a reliable method for counting Circulating Tumor Cells (CTC) using flow Cytometry. Flow Cytometry is a  method  of enumerating and examining minute particles suspended in a  fluid  when passed through an electronic detector. The system has a disposable chip. This chip checks for cross contamination  collect  analyzed sample and to freely measurement. CTC is  salient  biomarkers for so many cancers. There are many systems for enumeration based on either EpCAM/CD326 which  express  tumor cell before microscope or  RT-PCR. Protocols for this system can be applied onto other systems. Cultured cancer cells spiked into normal blood got enriched with  MACR  EpCAM  microbeads then  labeled  with APC instead of intracellular staining of cytokeratins.  EpCAM  allows enumeration of  intact  CTC, cellular integrity  maintenance  and concomitant  performance. Combination of  fineà ‚  tuned CTC and cytometric multicolor resulted into linear relationship between input and output  cell  count from zero to hundred of cells. Anti CD45  mAb  was used  to  give  satisfactory  signal/ noise ratio by  gate  exclusion of white blood cells  signal. There is little  influence  on lungs cancer cell PC-9 viability. CTC is of greater importance because it provides stratification of Anti-tumor treatment and furthering characterization. Several researchers have shown that circulating Tumor cells (CTC) in peripheral blood are significant prognostic marker for cancer (1-5). Presence of circulating tumor cells in the peripheral blood of patients  has been involved  in the Tumor  development  and metastasis  advancement. Response of  therapy  and evaluation of  disease  get  predicted  by change in circulating tumor cells. Several methods  have been used  in the  CTC-enrichment  and  discovery, but the  standard  metho d is the FDA-approved cell search system (Veridex) (Takao, M., Takeda, K., 2011). This employs a 7.5ml of blood and involves epithelial cell adhesion molecules (EpCAM  /CD360) (8)-conjugated  immuno-magnetic  enrichment preceded by cell imaging  process  using  positive  immuno-staining  of  cytokenins. Later negative immunostaining of leucocyte common antigen (CD45) and DNA staining with  DAPI. The overall advantage of this method is the  rapid  read out of routine measurements.  This is due to the fact that  sizeable  information gets included  in the  data  and its capability of multicolor analysis.  This  method  also offers  precise  detection limit of  pure  cells of approximately (10^-5). Related research Benjamin and Steven conducted research on flow Cytometry. They inferred that there has been progress in  immuno-magnetic  and  flow  cytometry. Benjamin and Steven concluded that  flow  cytometry and immunomagneti c can detect and characterize circulating tumor cells. They  infer  that flow cytometry has demonstrated prognostic  importance  in prostate and breast cancer. In Benjamin’s and Steven article about â€Å" circulating tumor cells in colorectal cancer †¦Ã¢â‚¬  there are reviews regarding the  historical  and  development  information about  identification  and enumeration of circulating tumor cells in colorectal cancer. The presence of circulating tumor cells in patients having metastatic carcinomas get  linked  with poor survival predictions (Tych,  Frederik,  Sjoerd,  Joost, Jan  &Leon, 2011). According to their article based on research, image cytometer,  cell  tracks got  developed  to  advance  the enumeration of rare circulating tumor cells. Cell search  system  got used to  enumerate  circulating tumor cells in seven point five milliliters (7.5 Ml) of  bold  of nine healthy controls and sixty eight patients. The results  were obtained  from cell search  system  were analyzed again using image cytometer. Then automated categorization of events  was executed  by random forest  process  using