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A series of flexible design adaptations to the Nikon E‐C1 and E‐C2 confocal microscope systems for UV, multiphoton and FLIM imaging
Author(s) -
BOTCHWAY STANLEY W.,
SCHERER KATHRIN M.,
HOOK STEVE,
STUBBS CHRISTOPHER D.,
WESTON ELEANOR,
BISBY ROGER H.,
PARKER ANTHONY W.
Publication year - 2015
Publication title -
journal of microscopy
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.569
H-Index - 111
eISSN - 1365-2818
pISSN - 0022-2720
DOI - 10.1111/jmi.12218
Subject(s) - microscope , confocal , ultraviolet , optics , microscopy , fluorescence , materials science , laser , photobleaching , fluorescence lifetime imaging microscopy , confocal microscopy , excitation , optoelectronics , physics , quantum mechanics
Summary Multiphoton microscopy is widely employed in the life sciences using extrinsic fluorescence of low‐ and high‐molecular weight labels with excitation and emission spectra in the visible and near infrared regions. For imaging of intrinsic and extrinsic fluorophores with excitation spectra in the ultraviolet region, multiphoton excitation with one‐ or two‐colour lasers avoids the need for ultraviolet‐transmitting excitation optics and has advantages in terms of optical penetration in the sample and reduced phototoxicity. Excitation and detection of ultraviolet emission around 300 nm and below in a typical inverted confocal microscope is more difficult and requires the use of expensive quartz optics including the objective. In this technical note we describe the adaptation of a commercial confocal microscope (Nikon, Japan E‐C1 or E‐C2) for versatile use with Ti‐sapphire and OPO laser sources and the addition of a second detection channel that enables detection of ultraviolet fluorescence and increases detection sensitivity in a typical fluorescence lifetime imaging microscopy experiment. Results from some experiments with this setup illustrate the resulting capabilities.