Photobleaching Techniques/ FRAP-Fluorescence Recovery After Photobleaching
About this Technology
Photobleaching techniques are used as a strategy to reveal the diffusion/binding dynamics of molecules or the exchange between compartments in live cells, tissues or organisms. The most straightforward photobleaching technique is the Fluorescence Recovery After Photobleaching (FRAP). In FRAP, fluorescent molecules are deactivated in a region of interest by focused light (tipically a laser beam) and the recovery of the fluorescence intensity in that region is followed in time, quantified by suitable numerical analysis (normalization and curve fitting), and compared to the initial conditions.
Fluorescence recovery into the region is due to the diffusion of the molecules in and out the region. Bleached molecules, which can diffuse out of the region, are substituted by other unbleached molecules that diffuse inside the region from surrounding areas. Quantification and analysis of fluorescence recovery provide information of descriptive parameters such as mobile and immobile fraction and half time of recovery.
Further analysis by kinetic modelling provides information on the molecular dynamics such as the diffusional properties of the studied molecule and its binding characteristics.
Labs Using this Technology
Advanced Optical Microscopy
Universitat de Barcelona · Barcelona
Advanced Optical Microscopy Service - Nanoimaging Unit
Centro Andaluz de Nanomedicina y Biotechnología (BIONAND) · Málaga
Flow Cytometry and Confocal Microscopy Lab
IRB Lleida-Universitat de Lleida · Lérida
Microscopia Confocal SIdI-UAM
Universidad Autónoma de Madrid (UAM) · Madrid
Microscopy Core Facility
Universidad de Murcia · Murcia
Microscopy and Dynamic Imaging Unit
Instituto de Salud Carlos III · Madrid
Optical spectroscopy and microscopy plaftorm
CIC biomaGUNE · San Sebastián
Plataforma Cientifico-Tecnica de Microscopia Confocal
Instituto de Investigacion Biomedica Sant Pau · Barcelona
Servicio de Microscopia
INSTITUTO DE PARASITOLOGIA Y BIOMEDICINA LOPEZ NEYRA · Granada
Servicio de Microscopia Confocal y Citometria de Flujo
cicCartuja-Centro de Investigaciones Cientificas Isla de la Cartuja (CSIC-US) · Sevilla
Servicio de Microscopia Optica Avanzada IBBTEC
Consejo Superior de Investigaciones Cientificas (CSIC) · Santander
Servicio de Microscopia Optica y Confocal
Consejo Superior de Investigaciones Cientificas (CSIC) · Madrid
Servicio de Microscopía
Consejo Superior de Investigaciones Cientificas (CSIC) · Salamanca
Servicio de Microscopía y Análisis de Imagen
Servicio de salud de Castilla-La Mancha (SESCAM) · Toledo
Servicio de Tecnicas Aplicadas a las Biociencias
Universidad de Extremadura · Badajoz
Unidad de Microscopia Optica Avanzada y Análisis de Imagen
FRAP/FLIP
Consejo Superior de Investigaciones Cientificas (CSIC) · Sevilla
Unidad de Microscopia e Imagen Molecular
FPS-Fundacion Progreso y Salud · Granada
Unidad de Microscopía
Consejo Superior de Investigaciones Cientificas (CSIC) · Sevilla