1. Epi-illumination gradient light interference microscopy for imaging opaque structures
- Author
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Gabriel Popescu, Drew N. Robson, Kathryn Michele Sullivan, Ghazal Naseri Kouzehgarani, Eun Jung Min, Mikhail E. Kandel, Chenfei Hu, Martha U. Gillette, Catherine Best-Popescu, Hyunjoon Kong, and Jennifer M. Li
- Subjects
0301 basic medicine ,Microscope ,Materials science ,Opacity ,Science ,Phase-contrast microscopy ,General Physics and Astronomy ,01 natural sciences ,Article ,General Biochemistry, Genetics and Molecular Biology ,Light scattering ,Interference microscopy ,law.invention ,Tendons ,010309 optics ,Mice ,03 medical and health sciences ,Imaging, Three-Dimensional ,Optics ,law ,0103 physical sciences ,Microscopy ,Animals ,Humans ,Microscopy, Interference ,lcsh:Science ,Zebrafish ,Neurons ,Multidisciplinary ,Birefringence ,Scattering ,business.industry ,Optical Imaging ,Brain ,Imaging and sensing ,Hep G2 Cells ,Quartz ,General Chemistry ,Rats ,030104 developmental biology ,Semiconductors ,Larva ,Inverse scattering problem ,lcsh:Q ,business ,HeLa Cells - Abstract
Multiple scattering and absorption limit the depth at which biological tissues can be imaged with light. In thick unlabeled specimens, multiple scattering randomizes the phase of the field and absorption attenuates light that travels long optical paths. These obstacles limit the performance of transmission imaging. To mitigate these challenges, we developed an epi-illumination gradient light interference microscope (epi-GLIM) as a label-free phase imaging modality applicable to bulk or opaque samples. Epi-GLIM enables studying turbid structures that are hundreds of microns thick and otherwise opaque to transmitted light. We demonstrate this approach with a variety of man-made and biological samples that are incompatible with imaging in a transmission geometry: semiconductors wafers, specimens on opaque and birefringent substrates, cells in microplates, and bulk tissues. We demonstrate that the epi-GLIM data can be used to solve the inverse scattering problem and reconstruct the tomography of single cells and model organisms., Quantitative phase imaging techniques have been limited by multiple scattering of light or its use in transmission mode. Here, the authors show a gradient light interference microscopy method in a reflection geometry which allows for label-free phase imaging of bulk and opaque samples.
- Published
- 2019