Phase-sensitive, angle-resolved light-scattering microscopy of single cells
Autor: | Andrew J. Berger, Robert L. Draham, Kaitlin J. Dunn |
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Rok vydání: | 2020 |
Předmět: |
Materials science
Light Mie scattering Fast Fourier transform 02 engineering and technology 01 natural sciences Light scattering 010309 optics Mice chemistry.chemical_compound Optics Planar Phagocytosis 0103 physical sciences Microscopy Animals Scattering Radiation Microscopy Phase-Contrast Cells Cultured Thymocytes Fourier Analysis business.industry Scattering Macrophages Equipment Design 021001 nanoscience & nanotechnology Atomic and Molecular Physics and Optics Mice Inbred C57BL chemistry Polystyrenes Polystyrene 0210 nano-technology business Beam (structure) |
Zdroj: | Optics Letters. 45:6775 |
ISSN: | 1539-4794 0146-9592 |
Popis: | We report what is to our knowledge the first use of Fourier phase microscopy (FPM) to estimate diameters of individual single-micrometer beads and to classify cells based upon changes in scatterer size distribution. FPM, a quantitative phase imaging (QPI) method, combines the planar illumination typically used in off-axis QPI (ideal for Mie theory analysis) with the common-path geometry typically used in on-axis QPI (ideal for optimizing angular scattering range). Low-spatial-frequency imaging artifacts inherent to FPM have negligible impact upon these angular-domain applications. The system is simple to align and stable, and requires no external reference beam. Angular scattering patterns obtained from single 1 µm polystyrene beads in glycerol ( Δ n = 0.11 ) display unprecedented fidelity to Mie theory, produce diameter estimates consistent with the manufacturer’s specifications, and offer precision on the scale of tens of nanometers. Measurements of macrophages at different stages of antibody-dependent cellular phagocytosis demonstrate the ability to detect changes in a cell’s scattering caused by the presence of phagocytosed material within. |
Databáze: | OpenAIRE |
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