Open AccessOptical sectioning of unlabeled samples using bright-field microscopy
Author(s) -
Braulio Gutiérrez–Medina
Publication year - 2022
Publication title -
proceedings of the national academy of sciences of the united states of america
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.011
H-Index - 771
eISSN - 1091-6490
pISSN - 0027-8424
DOI - 10.1073/pnas.2122937119
Subject(s) - microscopy , optical sectioning , optical microscope , light sheet fluorescence microscopy , microscope , fluorescence microscope , optics , materials science , bright field microscopy , instrumentation (computer programming) , depth of field , fluorescence , computer science , scanning confocal electron microscopy , physics , scanning electron microscope , operating system
The bright-field (BF) optical microscope is a traditional bioimaging tool that has been recently tested for depth discrimination during evaluation of specimen morphology; however, existing approaches require dedicated instrumentation or extensive computer modeling. We report a direct method for three-dimensional (3D) imaging in BF microscopy, applicable to label-free samples, where we use Köhler illumination in the coherent regime and conventional digital image processing filters to achieve optical sectioning. By visualizing fungal, animal tissue, and plant samples and comparing with light-sheet fluorescence microscopy imaging, we demonstrate the accuracy and applicability of the method, showing how the standard microscope is an effective 3D imaging device.