Effect of Photobleaching on the Viability of Hamster Fibroblast NIL-8M2 Cells
MetadataShow full item record
Fluorescence recovery after photobleaching (FRAP) is a technique employed to estimate diffusion coefficients of membraneous components. Fluorescent labels, when exposed to high intensity light and molecular oxygen, are irreversibly quenched or photobleached. This project examined the possibility of photodamage to the cell caused by the photobleaching light, and sensitized by the fluorescent label. The parameter used to quantitate photodamage was cell viability. Cell viability was evaluated 48 hours after photobleaching either a spot (1.6 urn) or the entire cell with variable light dosages (dosage = intensity of light x time of exposure). An elevation in cell death was found after photobleaching, increasing with the dosage of light. Photobleaching of the 1.6 urn spot resulted. in a higher death percentage than applying the same dosage to the entire cell. Significant meaning cannot be attached to these results, however, due to a large number of complications encountered during this project.