Print Email Facebook Twitter Multi-line fluorescence scanning microscope for multi-focal imaging with unlimited field of view Title Multi-line fluorescence scanning microscope for multi-focal imaging with unlimited field of view Author van der Graaff, L. (TU Delft ImPhys/Quantitative Imaging) van Leenders, Geert J. L. H. (Erasmus MC) Boyaval, Fanny (Leiden University Medical Center) Stallinga, S. (TU Delft ImPhys/Imaging Physics) Department ImPhys/Imaging Physics Date 2019 Abstract Confocal scanning microscopy is the de facto standard modality for fluorescence imaging. Point scanning, however, leads to a limited throughput and makes the technique unsuitable for fast multi-focal scanning over large areas. We propose an architecture for multi-focal fluorescence imaging that is scalable to large area imaging. The design is based on the concept of line scanning with continuous ‘push broom’ scanning. Instead of a line sensor, we use an area sensor that is tilted with respect to the optical axis to acquire image data from multiple depths inside the sample simultaneously. A multi-line illumination where the lines span a plane conjugate to the tilted sensor is created by means of a diffractive optics design, implemented on a spatial light modulator. In particular, we describe a design that uses higher order astigmatism to generate focal lines of substantially constant peak intensity along the lines. The proposed method is suitable for fast 3D image acquisition with unlimited field of view, it requires no moving components except for the sample scanning stage, and provides intrinsic alignment of the simultaneously scanned focal slices. As proof of concept, we have scanned 9 focal slices simultaneously over an area of 36 mm2 at 0.29 µm pixel size in object space. The projected ultimate throughput that can be realized with the proposed architecture is in excess of 100 Mpixel/s To reference this document use: http://resolver.tudelft.nl/uuid:00b32681-f989-42c2-9b7c-36ec6262f657 DOI https://doi.org/10.1364/BOE.10.006313 ISSN 2156-7085 Source Biomedical Optics Express, 10 (12), 6313-6339 Part of collection Institutional Repository Document type journal article Rights © 2019 L. van der Graaff, Geert J. L. H. van Leenders, Fanny Boyaval, S. Stallinga Files PDF boe_10_12_6313.pdf 1.99 MB Close viewer /islandora/object/uuid:00b32681-f989-42c2-9b7c-36ec6262f657/datastream/OBJ/view