Working with the cells
Today David invited me be something like a witness of an experiment concerning the Zstk drug. It was very interesting. The experiment can be summarized as follows:
- The marked cells comes from a human kidney ephitelium. They were marked with two different fluorescent protein. A red fluorescent protein was used to mark the nucleus of the cell and a red fluorescent protein (genetically introduced in the cell) was used to follow the dynamical behavior of the cell. The red protein correspond is a histone (I must look for some information concerning how histones work in the nucleus of the cell).
- The marked cells were put in a glass recipient (to make them stay with no motion when using microscopy).
- The drug was applied to the cells.
- A robotic optical Nikon microscope was programmed (with a openware Java application) to take photos of the cells from 5 different locations each two minutes during two hours.
- Once the photos were recorded, the cell were thoroughly washed to eliminate the drug using the PBS solution.
- The washed cells were put again in the microscope and the drug was used again.
The purpose of the experiments is to verify if the hysteresis behavior is also detected in this kind of cells.
- A Matlab script will be applied to recover from the photos of the cells the evolution of the FOXO1a protein (the script will detect how FOXO1a enters the nucleus of the cell, via the comparison between the changing levels of red and green protein inthe nucleus -the drug action is translated in terms of motion of the green protein from the cytoplasm to the nucleus-).
I want one this robotic microscopes!
Discussing models with David
The two feedback loops must be argued as a qualitative explanation of the phenomenon (a complementary experiment must be performed to see if a modification of the network allows the detection of the counterclockwise hysteretical phenomenon).