title: Optics clustered to output unique solutions: a multi-laser facility for combined single molecule and ensemble microscopy.
abstract: Optics clustered to output unique solutions (OCTOPUS) is a microscopy platform that combines single molecule and ensemble imaging methodologies. A novel aspect of OCTOPUS is its laser excitation system, which consists of a central core of interlocked continuous wave and pulsed laser sources, launched into optical fibres and linked via laser combiners. Fibres are plugged into wall-mounted patch panels that reach microscopy end-stations in adjacent rooms. This allows multiple tailor-made combinations of laser colours and time characteristics to be shared by different end-stations minimising the need for laser duplications. This setup brings significant benefits in terms of cost effectiveness, ease of operation, and user safety. The modular nature of OCTOPUS also facilitates the addition of new techniques as required, allowing the use of existing lasers in new microscopes while retaining the ability to run the established parts of the facility. To date, techniques interlinked are multi-photon/multicolour confocal fluorescence lifetime imaging for several modalities of fluorescence resonance energy transfer (FRET) and time-resolved anisotropy, total internal reflection fluorescence, single molecule imaging of single pair FRET, single molecule fluorescence polarisation, particle tracking, and optical tweezers. Here, we use a well-studied system, the epidermal growth factor receptor network, to illustrate how OCTOPUS can aid in the investigation of complex biological phenomena.
authors.affiliation: Science and Technology Facilities Council, Central Laser Facility, Research Complex at Harwell, Rutherford Appleton Laboratory, Didcot, Oxford OX11 0FA, United Kingdom.
authors.forename: David T
authors.initials: DT
authors.lastname: Clarke
authors.forename: Stanley W
authors.initials: SW
authors.lastname: Botchway
authors.forename: Benjamin C
authors.initials: BC
authors.lastname: Coles
authors.forename: Sarah R
authors.initials: SR
authors.lastname: Needham
authors.forename: Selene K
authors.initials: SK
authors.lastname: Roberts
authors.forename: Daniel J
authors.initials: DJ
authors.lastname: Rolfe
authors.forename: Christopher J
authors.initials: CJ
authors.lastname: Tynan
authors.forename: Andrew D
authors.initials: AD
authors.lastname: Ward
authors.forename: Stephen E D
authors.initials: SE
authors.lastname: Webb
authors.forename: Rahul
authors.initials: R
authors.lastname: Yadav
authors.forename: Laura
authors.initials: L
authors.lastname: Zanetti-Domingues
authors.forename: Marisa L
authors.initials: ML
authors.lastname: Martin-Fernandez
mesh_terms: D000818:Animals; D002460:Cell Line; D002470:Cell Survival; D003116:Color; D066246:ErbB Receptors / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D007834:Lasers; D008853:Microscopy / Q000295:instrumentation*; D055620:Optical Phenomena; D017785:Photons; D011487:Protein Conformation; D021381:Protein Transport; D015398:Signal Transduction
chemical_list: D066246:ErbB Receptors