Fluorescence in C. Elegans: Discussion

1. One way to control for auto-fluorescence in C. elegans is to observe younger nematodes. This is because the auto-fluorescing molecules that exist in C. elegans are lipofuscins, which is in the family known as “aging pigments”. Lipofuscins accumulate in nematodes as a by-product of oxidative stress. Therefore younger nematodes will have a lot less lipofuscins then older nematodes. One way to control auto-fluorescence related to filter cubes and fluorescence microscopy is to use a filter cube that will block out the fluorescence caused by lipofuscins. This can be done by choosing a filter cube that does not excite within the excitation range of 300-400nm for lipofuscins. This would elimintate any auto-fluorescence within the specimen.
2. The molecular quality that accounts for the broad emission spectra of lipofuscins is that they are made up of non-functional proteins that are cross-linked. The proteins become cross-linked due to oxidative processes that occur in nearby systems. Lipofuscins share the quality of having a broad emission spectrum with other cross-linked proteins.
3. -
   1. It would be possible to conclude whether receptor-mediated endocytosis is occurring based on the contents of the vesicles. If the contents of the vesicles are homogeneous then it would make sense that the mechanism used for vesicle formation is selective. Therefore it could be selective through receptor-mediated endocytosis and phagocytosis. It the contents of the vesicles are a heterogeneous mixture then the mechanism used for vesicle formation would most likely be non-selective such as pinocytosis.
   2. Based on my observations I am unable to conclude whether or not C. elegans is limited to using cellular endocytosis or any other digestive mechanism to incorporate large digestive stimuli. This is because in my observations the fluorescence caused by the FITC-microshpere treatment seemed to be limited to the intestines. This makes it unclear if the nematodes where still in the process of incorporating the microspheres or if the microspheres are merely passing through undigested.

Fluorescence in C. elegans Introduction

Fluorescence in C. elegans Methods

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