Team:UFSCar-Brasil/part1.html
Plasmolysis
How Bug Shoo works
Fitting Data
From the experimental data presented in Table 1 on the appendix, we constructed a 3D graph (as shown in Figure 1) to better understanding of the system, which is the relationship between absorbance, time and PEG concentration.
![](https://static.igem.org/mediawiki/2015/8/87/UFSCar-Brasil_abs_t_peg.jpg)
Graph 1: Experimental dots of absorbance versus time versus PEG concentration.
With the graph in hand, our next step was to find the surface that best fits the experimental points. For this we use the parameter r² , because that shows us the percentage of points that fit a certain curve, in other words, what is the best curve that most closely approximates the behavior of the data. And thus, we conclude that this was:
z= \(z_0\)+ax+by+cx²+dy²+fxy.
This curve obtained r² = 0.95457, what means that 95% of data points are modeled by this function. Using the OringiPro8 software to fitting the function, we also finds the value of the constants, which can be seen in Table 2 of the appendix.
From the known curve was possible to simulate the surface that models the most likely behavior of the three variables in question.
![](https://static.igem.org/mediawiki/2015/f/f4/UFSCar-Brasil_fitted.jpg)
Graph 2: Fitted surface obtained using the expression (1).
Results and Discussion
With the ultimate goal to find the main points of the system, meaning those points where his plane tangent in the surface has gradient value null. But first, we analyzed (using the Hessian matrix) on these important points to make possible to know whether these are the maximum points, minimum or saddle point of the function.
The calculation of the Hessian matrix is given by:
thus to find the value of the Hessian of z(x,y) we obtained the partial derivatives of the second order, whose values are below:
As H[f(x,y)]<0, we know that in this function there is a saddle point and there are no minimum or maximum values (Guidorizzi, 1997). Saddle point is understood as the point where the slope is zero, but it is not a maximum or minimum value. In it there is the highest elevation in one direction and lowest in the perpendicular direction.
Now, known the point we want to find, we take the partial derivatives of the first order function and made them equal to zero. To know:
replacing the values that are found in Table 2 of the appendix, we have that x=(2da-bf)/(f²-4cd)=1,5 and thus y=22.2 and z=0.63 . Then it is possible to concluded that for optimal absorbance point (z = 0.63), we know that it is necessary 22.2% PEG where it gets 1.5 weeks of plasmolysis.
Conclusions
Through these analyzes we could first find one function that suited in r² = 95%, which is very satisfactory, and validates our whole discussion about the obtained surface.
Then, through the Hessian we found a saddle point, which is also particularly relevant biologically. Because the absorbance measures the metabolic activity of bacteria, and how we want that they are plasmolysed, this value should be low, but not zero, because if so, they would be dead and would not serve for our propose. As it is a saddle point, we know that it is not null. Finally, the point was found through gradient of the function and we concluded that for the minimum value of absorbance the best PEG concentration is 22,2%.
Appendix
List of tables
Table 1: Experimental dates obtained in microbiology lab
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