TY - JOUR T1 - LITE microscopy: a technology for high numerical aperture, low photobleaching fluorescence imaging JF - bioRxiv DO - 10.1101/181644 SP - 181644 AU - TC Fadero AU - TM Gerbich AU - K Rana AU - A Suzuki AU - M DiSalvo AU - KN Schaefer AU - JK Heppert AU - TC Boothby AU - B Goldstein AU - M Peifer AU - NL Allbritton AU - AS Gladfelter AU - AS Maddox AU - PS Maddox Y1 - 2017/01/01 UR - http://biorxiv.org/content/early/2017/10/04/181644.abstract N2 - 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. ER -