Team:Jiangsu High School/Measurement
Best Measurement
Best Measurement Award
The main aim of our project is to figure out which compounds are best at instigating cell autophagy in order to find compounds that have potential in stopping the accumulation of harmful protein in the brain. We have used a confocal microscope to observe our samples during the autophagy flux test. The plasmid DNA we inserted into the cells was mTagRFP-mWasabi-LC3 and would indicate the effectiveness of the autophagy. The advantage this plasmid DNA has on different techniques is considerable—for example, electron microscopy is not clear enough to express findings at a molecular level and the Western blot test is not completely reliable for examining protein. mTagRFP-mWasabi-LC3 combines LC3 with the fluorescent protein mWasabi which gives us the ability to examine autophagy dynamically. During the fluorescence test, autophagosomes would be seen as having yellow fluorescence while autolysosomes would have red fluorescence. In comparing the amount of fluorescence in each sample, we were able to recognize C4 and C13 as the most luminous—and thus the most effective as catalysts for autophagy (Fig 1a; Fig 1b).
