Red Española de Microscopía Óptica Avanzada
Photobleaching Techniques/ FRAP-Fluorescence Recovery After Photobleaching
 
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Description
 
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.
This technique at work
 
Universitat de Barcelona, CCiTUB- Centres Cientifics i Tecnologics - Advanced Optical Microscopy Unit
 
cicCartuja-Centro de Investigaciones Cientificas Isla de la Cartuja (CSIC-US), IBVF-Instituto de Bioquimica Vegetal y Fotosintesis - Servicio de Microscopia Confocal y Citometria de Flujo
 
Consejo Superior de Investigaciones Cientificas (CSIC), Centro Nacional de Biotecnologia (CNB-CSIC) - Servicio de Microscopia Confocal del Centro Nacional de Biotecnologia
 
Consejo Superior de Investigaciones Cientificas (CSIC), Centro Andaluz de Biologia del Desarrollo - Unidad de Microscopia Optica
 
FPS-Fundacion Progreso y Salud, GENYO-Centro Pfizer-Universidad de Granada-Junta de Andalucia de Genomica e Investigacion Oncologica - Unidad de Microscopia e Imagen Molecular
 
Consejo Superior de Investigaciones Cientificas (CSIC), Centro de Biología Molecular Severo Ochoa - Servicio de Microscopia Optica y Confocal
 
Consejo Superior de Investigaciones Cientificas (CSIC), Instituto de Biomedicina y Biotecnologia de Cantabria - IBBTEC - Servicio de Microscopia Optica Avanzada IBBTEC
 
Consejo Superior de Investigaciones Cientificas (CSIC), Instituto de Biología Funcional y Genómica IBFG - Servicio de Microscopía
 
Universidad Autónoma de Madrid (UAM), Servicio Interdepartamental de Investigación (SIdI) UAM - Microscopia Confocal SIdI-UAM
 
Centro Andaluz de Nanomedicina y Biotechnología (BIONAND), Unidad de Nanoimagen - Advanced Optical Microscopy Service - Nanoimaging Unit
 
Instituto de Salud Carlos III, Centro Nacional de Investigaciones Cardiovasculares (CNIC) - Microscopy and Dynamic Imaging Unit
 
Instituto de Investigacion Biomedica Sant Pau, Instituto de Investigacion Sant Pau - Plataforma Cientifico-Tecnica de Microscopia Confocal
 
INSTITUTO DE PARASITOLOGIA Y BIOMEDICINA LOPEZ NEYRA, INSTITUTO DE PARASITOLOGIA Y BIOMEDICINA LOPEZ NEYRA - Servicio de Microscopia
 
INSTITUTO DE PARASITOLOGIA Y BIOMEDICINA LOPEZ NEYRA, INSTITUTO DE PARASITOLOGIA Y BIOMEDICINA LOPEZ NEYRA - Servicio de Microscopia
 
Universidad de Extremadura, Servicios de Apoyo a la Investigación - Servicio de Tecnicas Aplicadas a las Biociencias
 
IRB Lleida-Universitat de Lleida, IRB Lleida-Universitat de Lleida - Flow Cytometry and Confocal Microscopy Lab
 
Servicio de salud de Castilla-La Mancha (SESCAM), Hospital Nacional de Parapléjicos - Servicio de Microscopía y Análisis de Imagen
 
Institut d'Investigació Biomèdica de Bellvitge (IDIBELL), Centre de Medicina Regenerativa de Barcelona (CMRB) - Optical Microscopy Unit
 
 
Related training
 
 
Type Document Dates Lab
 
Related material
 
 
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