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Fast Optical Sectioning for Widefield Fluorescence Mesoscopy with the Mesolens based on HiLo Microscopy.
Schniete, Jan; Franssen, Aimee; Dempster, John; Bushell, Trevor J; Amos, William Bradshaw; McConnell, Gail.
Afiliação
  • Schniete J; Department of Physics, University of Strathclyde, Glasgow, G4 0NG, United Kingdom. jan.schniete@strath.ac.uk.
  • Franssen A; Strathclyde Institute of Pharmacy and Biomedical Sciences, University of Strathclyde, Glasgow, G4 0RE, United Kingdom.
  • Dempster J; Strathclyde Institute of Pharmacy and Biomedical Sciences, University of Strathclyde, Glasgow, G4 0RE, United Kingdom.
  • Bushell TJ; Strathclyde Institute of Pharmacy and Biomedical Sciences, University of Strathclyde, Glasgow, G4 0RE, United Kingdom.
  • Amos WB; Department of Physics, University of Strathclyde, Glasgow, G4 0NG, United Kingdom.
  • McConnell G; MRC Laboratory of Molecular Biology, Cambridge Biomedical Campus, Cambridge, CB2 OQH, United Kingdom.
Sci Rep ; 8(1): 16259, 2018 11 02.
Article em En | MEDLINE | ID: mdl-30390029
We present here a fast optical sectioning method for mesoscopy based on HiLo microscopy, which makes possible imaging of specimens of up to 4.4 mm × 3 mm × 3 mm in volume in under 17 hours (estimated for a z-stack comprising 1000 images excluding computation time) with subcellular resolution throughout. Widefield epifluorescence imaging is performed with the Mesolens using a high pixel-number camera capable of sensor-shifting to generate a 259.5 Megapixel image, and we have developed custom software to perform HiLo processing of the very large datasets. Using this method, we obtain comparable sectioning strength to confocal laser scanning microscopy (CLSM), with sections as thin as 6.8 ± 0.2 µm and raw acquisition speed of 1 minute per slice which is up to 30 times faster than CLSM on the full field of view (FOV) of the Mesolens of 4.4 mm with lateral resolution of 0.7 µm and axial resolution of 7 µm. We have applied this HiLo mesoscopy method to image fixed and fluorescently stained hippocampal neuronal specimens and a 5-day old zebrafish larva.

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Idioma: En Revista: Sci Rep Ano de publicação: 2018 Tipo de documento: Article País de afiliação: Reino Unido

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Idioma: En Revista: Sci Rep Ano de publicação: 2018 Tipo de documento: Article País de afiliação: Reino Unido