RT Journal Article
SR Electronic
T1 LITE microscopy: a technology for high numerical aperture, low photobleaching fluorescence imaging
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 181644
DO 10.1101/181644
A1 TC Fadero
A1 TM Gerbich
A1 K Rana
A1 A Suzuki
A1 M DiSalvo
A1 KN Schaefer
A1 JK Heppert
A1 TC Boothby
A1 B Goldstein
A1 M Peifer
A1 NL Allbritton
A1 AS Gladfelter
A1 AS Maddox
A1 PS Maddox
YR 2017
UL http://biorxiv.org/content/early/2017/10/04/181644.abstract
AB Fluorescence microscopy is a powerful approach for studying sub-cellular dynamics at high spatiotemporal resolution; however, conventional fluorescence microscopy techniques are light-intensive and introduce unnecessary photodamage. Light sheet fluorescence microscopy (LSFM) mitigates these problems by selectively illuminating the focal plane of the detection objective using orthogonal excitation. Orthogonal excitation requires geometries that physically limit the detection objective numerical aperture (NA), thereby limiting both light-gathering efficiency (brightness) and native spatial resolution. We present a novel LSFM method: Lateral Interference Tilted Excitation (LITE), in which a tilted light sheet illuminates the detection objective focal plane without a sterically-limiting illumination scheme. LITE is thus compatible with any detection objective, including oil immersion, without an upper NA limit. LITE combines the low photodamage of LSFM with the benefits of high brightness, high resolution, coverslipbased objectives. We demonstrate LITE’s utility for imaging animal, fungal, and plant model organisms over many hours at high spatiotemporal resolution.