Dual light‐emitting diode‐based multichannel microscopy for whole‐slide multiplane, multispectral and phase imaging
Year: 2017
Type: article
Abstract: We report the development of a multichannel microscopy for whole-slide multiplane, multispectral and phase imaging. We use trinocular heads to split the beam path into 6 independent channels and employ a camera array for parallel data acquisition, achieving a maximum data throughput of approximately 1 gigapixel per second. To perform single-frame rapid autofocusing, we place 2 near-infrared light-emitting diodes (LEDs) at the back focal plane of the condenser lens to illuminate the sample from 2 different incident angles. A hot mirror is used to direct the near-infrared light to an autofocusing camera. For multiplane whole-slide imaging (WSI), we acquire 6 different focal planes of a thick specimen simultaneously. For multispectral WSI, we relay the 6 independent image planes to the same focal position and simultaneously acquire information at 6 spectral bands. For whole-slide phase imaging, we acquire images at 3 focal positions simultaneously and use the transport-of-intensity equation to recover the phase information. We also provide an open-source design to further increase the number of channels from 6 to 15. The reported platform provides a simple solution for multiplexed fluorescence imaging and multimodal WSI. Acquiring an instant focal stack without z-scanning may also enable fast 3-dimensional dynamic tracking of various biological samples.
Source: Journal of Biophotonics
Authors: Jun Liao, Zhe Wang, Zibang Zhang, Zichao Bian, Kaikai Guo, Aparna Nambiar, Yutong Jiang, Shaowei Jiang, Jingang Zhong, Michael Choma, Guoan Zheng
Institutions: University of Connecticut, Jinan University, Yale University
Language: English
Subfield: Atomic and Molecular Physics, and Optics
Field: Physics and Astronomy
Domain: Physical Sciences
Open Access status: green
Awards: G2452223239, IDBR TYPE B: Development of a $100 high-throughput whole slide imaging kit, Quantitative optical imaging of cilia-driven fluid flow, G7506190253, G751407715, Massively parallel interferometric confocal microscopy using degenerate lasers, G8768410814, Development of a new light source for parallel optical coherence tomograph, G9000147070, G9000329701
OpenAlex ID: W2605095952