Open Source High Content Analysis Utilizing Automated Fluorescence Lifetime Imaging Microscopy

Frederik G&#246;rlitz; Douglas J. Kelly; Sean C. Warren; Dominic Alibhai; Lucien West; Sunil Kumar; Yuriy Alexandrov; Ian Munro; Edwin Garcia; James McGinty; Clifford Talbot; Remigiusz A. Serwa; Emmanuelle Thinon; Vincenzo da Paola; Edward J. Murray; Frank Stuhmeier; Mark A. A. Neil; Edward W. Tate; Christopher Dunsby; Paul M. W. French

doi:10.3791/55119

Journal

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Biology

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Open Source High Content Analysis Utilizing Automated Fluorescence Lifetime Imaging Microscopy

JoVE Journal
Biology

This content is Free Access.

JoVE Journal Biology

Open Source High Content Analysis Utilizing Automated Fluorescence Lifetime Imaging Microscopy

11,963 Views

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09:30 min

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January 18, 2017

DOI:

10.3791/55119-v

DOI:

10.3791/55119-v

•

09:30 min

January 18, 2017

•

11941 Views

¹Photonics Group, Department of Physics,Imperial College London ²Institute for Chemical Biology, Department of Chemistry,Imperial College London ³MRC Clinical Sciences Centre,Hammersmith Hospital ⁴Chemical Biology Section, Department of Chemistry,Imperial College London ⁵Retroscreen Virology Ltd ⁶Pfizer Global Research and Development,Pfizer Limited, Sandwich, Kent, UK ⁷Centre for Histopathology,Imperial College London

Summary

We present an open source high content analysis (HCA) instrument utilizing automated fluorescence lifetime imaging (FLIM) for assaying protein interactions using Förster resonance energy transfer (FRET) based readouts. Data acquisition for this openFLIM-HCA instrument is controlled by software written in µManager and data analysis is undertaken in FLIMfit.