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Phase and fluorescence imaging with a surprisingly simple microscope based on chromatic aberration

  • Mandula, Ondřej
  • Kleman, Jean-Philippe
  • Lacroix, Françoise
  • Allier, Cédric
  • Fiole, Daniel
  • Hervé, Lionel
  • Blandin, Pierre
  • Kraemer, Dorothee
  • Morales, Sophie
Publication Date
Jan 20, 2020
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We propose a simple and compact microscope combining phase imaging withmulti-color fluorescence using a standard bright-field objective. The phase image of the sampleis reconstructed from a single, approximately 100μmout-of-focusimage taken under semi-coherent illumination, while fluorescence is recordedin-focusin epi-fluorescence geometry.The reproducible changes of the focus are achieved with specifically introduced chromaticaberration in the imaging system. This allows us to move the focal plane simply by changingthe imaging wavelength. No mechanical movement of neither sample nor objective or any otherpart of the setup is therefore required to alternate between the imaging modality. Due to itssmall size and the absence of motorized components the microscope can easily be used insidea standard biological incubator and allows long-term imaging of cell culture in physiologicalconditions. A field-of-view of 1.2 mm2allows simultaneous observation of thousands of cellswith micro-meter spatial resolution in phase and multi-channel fluorescence mode. In thismanuscript we characterize the system and show a time-lapse of cell culture in phase andmulti-channel fluorescence recorded inside an incubator. We believe that the small dimensions,easy usage and low cost of the system make it a useful tool for biological research.

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