Chew, Teng-Leong Cell Imaging Facility, Feinberg School of Medicine, Northwestern University, Chicago, Illinois.
- Photobleaching techniques
- Fluorescence recovery after photobleaching (FRAP)
- Fluorescence loss in photobleaching (FLIP)
- Photoswitch techniques
- Fluorescence resonance energy transfer (FRET) techniques
- Links to Primary Literature
- Additional Readings
Proteins, the molecules that perform the majority of biological functions in a cell, are in general colorless. Biologists conventionally use fluorescently tagged antibodies that specifically recognize their proteins of interest in order to indirectly visualize these biomolecules. Antibodies can be introduced into the cells by one of two methods: (1) permeabilization, wherein solvents, detergents, or drugs are used to form gaps in the plasma membrane that allow the passage of the antibodies; (2) microinjection, through which antibodies are introduced with a microneedle. In most cases, permeabilization kills the cells, and immunofluorescence of fixed cells generates only static images, providing a snapshot of otherwise highly dynamic biological structures and processes. On the other hand, microinjection will generate a high local protein concentration of antibodies that may affect the biological functions of the cells. In addition, microinjection is a laborious process that results in relatively few cells suitable for imaging.
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