Frapid: achieving full automation of FRAP for chemical probe validation

Clarence Yapp; Catherine Rogers; Pavel Savitsky; Martin Philpott; Susanne Müller

doi:10.1364/BOE.7.000422

Frapid: achieving full automation of FRAP for chemical probe validation

Biomed Opt Express. 2016 Jan 11;7(2):422-41. doi: 10.1364/BOE.7.000422. eCollection 2016 Feb 1.

Authors

Clarence Yapp¹, Catherine Rogers², Pavel Savitsky³, Martin Philpott⁴, Susanne Müller²

Affiliations

¹ Target Discovery Institute, NDMRB, University of Oxford, Oxford, OX3 7FZ, UK; Botnar Research Centre, NDORMS, University of Oxford, Oxford, OX3 7LD, UK.
² Target Discovery Institute, NDMRB, University of Oxford, Oxford, OX3 7FZ, UK.
³ Structural Genomics Consortium, ORCRB, University of Oxford, Oxford, OX3 7DQ, UK.
⁴ Botnar Research Centre, NDORMS, University of Oxford, Oxford, OX3 7LD, UK.

Abstract

Fluorescence Recovery After Photobleaching (FRAP) is an established method for validating chemical probes against the chromatin reading bromodomains, but so far requires constant human supervision. Here, we present Frapid, an automated open source code implementation of FRAP that fully handles cell identification through fuzzy logic analysis, drug dispensing with a custom-built fluid handler, image acquisition & analysis, and reporting. We successfully tested Frapid on 3 bromodomains as well as on spindlin1 (SPIN1), a methyl lysine binder, for the first time.

Keywords: (100.3008) Image recognition, algorithms and filters; (150.0150) Machine vision; (150.5758) Robotic and machine control; (170.1530) Cell analysis; (170.1790) Confocal microscopy; (170.2520) Fluorescence microscopy.