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In-phase-contrast microscopes, interference between light directly passing through a cell and light refracted by cellular components is used to create high-contrast, high-resolution images without staining. It is the oldest and simplest type of microscope that creates an image by altering the wavelengths of light rays passing through the specimen. Altered wavelength paths are created using an annular stop in the condenser. The annular stop produces a hollow cone of...
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Quantifying Microorganisms at Low Concentrations Using Digital Holographic Microscopy (DHM)
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Published on: November 1, 2017

Quantifying cellular differentiation by physical phenotype using digital holographic microscopy.

Kevin J Chalut1, Andrew E Ekpenyong, Warren L Clegg

  • 1Cavendish Laboratory, Department of Physics, University of Cambridge, Cambridge, CB3 0HE, UK. kc370@cam.ac.uk

Integrative Biology : Quantitative Biosciences From Nano to Macro
|January 21, 2012
PubMed
Summary
This summary is machine-generated.

Physical changes during cell differentiation are significant. Digital holographic microscopy (DHM) and optical stretching quantify these cellular changes, offering new insights into differentiation monitoring.

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Area of Science:

  • Biophysics
  • Cell Biology
  • Optical Microscopy

Background:

  • Cell differentiation involves well-documented biochemical shifts.
  • Significant, cell-wide physical changes during differentiation are less understood.
  • Quantifying physical changes aids in understanding and monitoring differentiation.

Purpose of the Study:

  • To investigate and quantify physical changes during myeloid precursor cell differentiation.
  • To establish biophotonic methods for marker-free monitoring of cell differentiation.
  • To determine if physical properties differ based on lineage commitment.

Main Methods:

  • Utilized Digital Holographic Microscopy (DHM), a marker-free quantitative phase microscopy technique.
  • Employed optical stretching, a laser trap-based marker-free technique, to assess cell softness.
  • Focused on differentiating myeloid precursor cells.

Main Results:

  • Significant changes in subcellular structure and refractive index were observed within one day of differentiation induction.
  • Distinct differences in physical properties were noted depending on the type of lineage commitment.
  • Significant changes in cell softness were detected within one day using optical stretching.

Conclusions:

  • DHM and optical stretching provide sensitive, marker-free methods to parameterize cellular differentiation.
  • These biophotonic techniques offer a way to quantify and understand cell differentiation with minimal perturbation.
  • The study highlights the importance of physical changes in cell differentiation and provides tools for their measurement.