Wednesday, November 20, 2019
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 toà establishà a reliable method for counting Circulating Tumor Cells (CTC) using flow Cytometry. Flow Cytometry is aà methodà of enumerating and examining minute particles suspended in aà fluidà when passed through an electronic detector. The system has a disposable chip. This chip checks for cross contaminationà collectà analyzed sample and to freely measurement. CTC isà salientà biomarkers for so many cancers. There are many systems for enumeration based on either EpCAM/CD326 whichà expressà tumor cell before microscope orà RT-PCR. Protocols for this system can be applied onto other systems. Cultured cancer cells spiked into normal blood got enriched withà MACRà EpCAMà microbeads thenà labeledà with APC instead of intracellular staining of cytokeratins.à EpCAMà allows enumeration ofà intactà CTC, cellular integrityà maintenanceà and concomitantà performance. Combination ofà fineà tuned CTC and cytometric multicolor resulted into linear relationship between input and outputà cellà count from zero to hundred of cells. Anti CD45à mAbà was usedà toà giveà satisfactoryà signal/ noise ratio byà gateà exclusion of white blood cellsà signal. There is littleà influenceà 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 patientsà has been involvedà in the Tumorà developmentà and metastasisà advancement. Response ofà therapyà and evaluation ofà diseaseà getà predictedà by change in circulating tumor cells. Several methodsà have been usedà in theà CTC-enrichmentà andà discovery, but theà standardà 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 (EpCAMà /CD360) (8)-conjugatedà immuno-magneticà enrichment preceded by cell imagingà processà usingà positiveà immuno-stainingà ofà cytokenins. Later negative immunostaining of leucocyte common antigen (CD45) and DNA staining withà DAPI. The overall advantage of this method is theà rapidà read out of routine measurements.à This is due to the fact thatà sizeableà information gets includedà in theà dataà and its capability of multicolor analysis.à Thisà methodà also offersà preciseà detection limit ofà pureà cells of approximately (10^-5). Related research Benjamin and Steven conducted research on flow Cytometry. They inferred that there has been progress inà immuno-magneticà andà flowà cytometry. Benjamin and Steven concluded thatà flowà cytometry and immunomagneti c can detect and characterize circulating tumor cells. Theyà inferà that flow cytometry has demonstrated prognosticà importanceà in prostate and breast cancer. In Benjaminââ¬â¢s and Steven article about ââ¬Å" circulating tumor cells in colorectal cancer â⬠¦Ã¢â¬ there are reviews regarding theà historicalà andà developmentà information aboutà identificationà and enumeration of circulating tumor cells in colorectal cancer. The presence of circulating tumor cells in patients having metastatic carcinomas getà linkedà with poor survival predictions (Tych,à Frederik,à Sjoerd,à Joost, Janà &Leon, 2011). According to their article based on research, image cytometer,à cellà tracks gotà developedà toà advanceà the enumeration of rare circulating tumor cells. Cell searchà systemà got used toà enumerateà circulating tumor cells in seven point five milliliters (7.5 Ml) ofà boldà of nine healthy controls and sixty eight patients. The resultsà were obtainedà from cell searchà systemà were analyzed again using image cytometer. Then automated categorization of eventsà was executedà by random forestà processà using
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