Introduction
According to Bio-Rad, Alamar Blue is a “cell viability assay reagent which contains the cell permeable, non-toxic and weakly fluorescent blue indicator dye called resazurin.” (Bio-Rad, n.d.). Meaning that the resazurin is going to be using an oxidation-reduction (REDOX) indicator that will undergo a color change due to the response of the cellular metabolic reduction.
“The reduced form resorufin is pink and highly fluorescent, and the intensity of fluorescence produced is proportional to the number of living cells respiring”, meaning that there will be a color change when introduced to a cell (Bio-Rad, n.d.). In more simple terms, the cell will have a color change due to the Alamar Blue penetrating the cell membrane making a …show more content…
Meaning where each dilution is going to take place and where to adding each substance into. In this case: Row A: was skipped completely, Row B: Columns 1-6 were for the untreated control cells (cells only in normal media), Row B: Columns 10-12 were for treated cells with plumbagin, Rows C and D: were for cells treated with doubling dilution of our reagent/chemical, and lastly Rows E and F: were for cells treated with doubling dilutions of the vehicle.
After knowing where each part of the experiment is taking place the experiment can start being conducted. All of the material were strayed with Ethanol (Aseptic Technique) before being placed inside the Fume Hood, included our gloves and the sleeves of the lab coats. After cleaning the hood with ethanol the materials were placed inside the hood, were we stated conducting the …show more content…
When using Alamar Blue each color change means a specific effect the well had. In this case, red means the cells are alive, purple means the cells are dying, and lastly blue means the cells have dead. Figure 1 results, demonstrated that all the wells that have cells have the color change of red/pink meaning that the cells are all alive. Demonstrating that the reagent/chemical (Curcumin) didn’t have any effect to the cells (Hek293A). As seen for Row B: Columns 10-12 the color change was to black meaning that the cells are dead. However, for those columns the cells were treated with plumbagin and demonstrated a normal reaction. This is because, “It is possible that the beneficial effect of plumbagin is due to the inhibition of NAD(P)H oxidase” (Ding, Chen, Lui, Che, Vetter, Chang, 2005). Meaning that it inhibits the growth of the cell, overall killing the cell. Since curcumin is an organic reagent/chemical a vehicle was added to be able to dissolve it (Popuri and Pagala, 2013). Making it able for the reagent/chemical be able to get inside the cell membrane and make the color. And as shown in Figure 1, the wells that have the vehicle had a color change of red/pink meaning that the cells are
According to Bio-Rad, Alamar Blue is a “cell viability assay reagent which contains the cell permeable, non-toxic and weakly fluorescent blue indicator dye called resazurin.” (Bio-Rad, n.d.). Meaning that the resazurin is going to be using an oxidation-reduction (REDOX) indicator that will undergo a color change due to the response of the cellular metabolic reduction.
“The reduced form resorufin is pink and highly fluorescent, and the intensity of fluorescence produced is proportional to the number of living cells respiring”, meaning that there will be a color change when introduced to a cell (Bio-Rad, n.d.). In more simple terms, the cell will have a color change due to the Alamar Blue penetrating the cell membrane making a …show more content…
Meaning where each dilution is going to take place and where to adding each substance into. In this case: Row A: was skipped completely, Row B: Columns 1-6 were for the untreated control cells (cells only in normal media), Row B: Columns 10-12 were for treated cells with plumbagin, Rows C and D: were for cells treated with doubling dilution of our reagent/chemical, and lastly Rows E and F: were for cells treated with doubling dilutions of the vehicle.
After knowing where each part of the experiment is taking place the experiment can start being conducted. All of the material were strayed with Ethanol (Aseptic Technique) before being placed inside the Fume Hood, included our gloves and the sleeves of the lab coats. After cleaning the hood with ethanol the materials were placed inside the hood, were we stated conducting the …show more content…
When using Alamar Blue each color change means a specific effect the well had. In this case, red means the cells are alive, purple means the cells are dying, and lastly blue means the cells have dead. Figure 1 results, demonstrated that all the wells that have cells have the color change of red/pink meaning that the cells are all alive. Demonstrating that the reagent/chemical (Curcumin) didn’t have any effect to the cells (Hek293A). As seen for Row B: Columns 10-12 the color change was to black meaning that the cells are dead. However, for those columns the cells were treated with plumbagin and demonstrated a normal reaction. This is because, “It is possible that the beneficial effect of plumbagin is due to the inhibition of NAD(P)H oxidase” (Ding, Chen, Lui, Che, Vetter, Chang, 2005). Meaning that it inhibits the growth of the cell, overall killing the cell. Since curcumin is an organic reagent/chemical a vehicle was added to be able to dissolve it (Popuri and Pagala, 2013). Making it able for the reagent/chemical be able to get inside the cell membrane and make the color. And as shown in Figure 1, the wells that have the vehicle had a color change of red/pink meaning that the cells are