Light needle microscopy with spatially transposed detection for axially resolved volumetric imaging

• Published in: Scientific reports Vol. 9; no. 1; pp. 11687 - 10
• Main Authors: Kozawa, Yuichi, Sato, Shunichi
• Format: Journal Article
• Language: English
• Published: England Nature Publishing Group 12.08.2019; Nature Publishing Group UK
• Subjects: Confocal microscopy; Fluorescence microscopy; Microscopy
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/s41598-019-48265-3

The demand for rapid three-dimensional volumetric imaging is increasing in various fields, including life science. Laser scanning fluorescence microscopy has been widely employed for this purpose; however, a volumetric image is constructed by two-dimensional image stacking with a varying observation plane, ultimately limiting the acquisition speed. Here we propose a method enabling axially resolved volumetric imaging without a moving observation plane in the framework of laser scanning microscopy. A scanning light needle spot with an extended focal depth provides excitation, which normally produces a deep focus image with a loss of depth information. In our method, the depth information is retrieved from transposed lateral information on an array detector by utilising non-diffracting and self-bending characteristics imposed on fluorescent signals. This technique, implemented in two-photon microscopy, achieves truly volumetric images constructed from a single raster scan of a light needle, which has the capability to significantly reduce the acquisition time.