BIOSC-101 Lecture Notes - Lecture 12: Total Internal Reflection, Confocal Microscopy, Scanning Electron Microscope

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The basic principle of fluorescence is that when something absorbs energy, it will have to come back. There are some states where they don"t radiate some energy but eventually it will. So in an epifluorescence microscope, we have this: A fluorescent light source will hit a filter to have some blue light and hit the specimen which will cause it to let out some green light. Immunohistochemistry is the method to label a protein with antibodies so it would be visible in immunofluorescence microscopy. Direct immunofluorescence is when we have a staining tag attached to the anti- body that attaches to a section of the cell. Indirect immunofluorescent is when one antibody attaches to a section of a cell and another anti-body attaches to the original antibody. There are two types of confocal microscopy: We use confocal microscopy to create an in-focus section through thick cells. This is when we restrict the focal plane of a fluorescent sample.

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