Standing-wave-excited multiplanar fluorescence in a laser scanning microscope reveals 3D information on red blood cells

Rumelo Amor, Sumeet Mahajan, William Bradshaw Amos, Gail McConnell

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Standing-wave excitation of fluorescence is highly desirable in optical microscopy because it improves the axial resolution. We demonstrate here that multiplanar excitation of fluorescence by a standing wave can be produced in a single-spot laser scanning microscope by placing a plane reflector close to the specimen. We report here a variation in the intensity of fluorescence of successive planes related to the Stokes shift of the dye. We show by the use of dyes specific for the cell membrane how standing-wave excitation can be exploited to generate precise contour maps of the surface membrane of red blood cells, with an axial resolution of 90 nm. The method, which requires only the addition of a plane mirror to an existing confocal laser scanning microscope, may well prove useful in studying diseases which involve the red cell membrane, such as malaria.
Original languageEnglish
Article number7359
Number of pages7
JournalScientific Reports
Publication statusPublished - 8 Dec 2014


  • fluorescence imaging
  • biological fluorescence
  • multiplanar excitation
  • laser scanning

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