A light sheet based high throughput 3D-imaging flow cytometer for phytoplankton analysis.

This paper reports a light sheet fluorescence imaging flow cytometer for 3D sectioning of phytoplankton. The instrument developed has the inherent advantages of high cell counting throughput and high spatial resolution information derived from flow cytometry and light sheet microscopy. The throughput of the instrument is quantified by the sample volume flow rate of 0.5 μl/min with a spatial resolution as achieved by light sheet microscopy. Preliminary results from 3D morphology of the internal chlorophyll-a structure of two dinoflagellates species show promising application potentials of the method for phytoplankton taxonomy of selected species and species groups.

[1]  Christoph J. Engelbrecht,et al.  Resolution enhancement in a light-sheet-based microscope (SPIM). , 2006, Optics letters.

[2]  Uros Krzic,et al.  Light sheet‐based fluorescence microscopy: More dimensions, more photons, and less photodamage , 2008, HFSP journal.

[3]  D. Stainier,et al.  Even fluorescence excitation by multidirectional selective plane illumination microscopy (mSPIM). , 2007, Optics letters.

[4]  G. Iannello,et al.  Confocal light sheet microscopy: micron-scale neuroanatomy of the entire mouse brain. , 2012, Optics express.

[5]  J. Swoger,et al.  Basic building units and properties of a fluorescence single plane illumination microscope. , 2007, The Review of scientific instruments.

[6]  Jan Huisken,et al.  Light sheet microscopy for real-time developmental biology. , 2011, Current opinion in genetics & development.

[7]  J. Lichtman,et al.  Optical sectioning microscopy , 2005, Nature Methods.

[8]  Zoe V. Finkel,et al.  Phytoplankton in a changing world: cell size and elemental stoichiometry , 2010 .

[9]  Emmanuel G. Reynaud,et al.  The future of three-dimensional microscopic imaging in marine biology , 2011 .

[10]  F. Del Bene,et al.  Optical Sectioning Deep Inside Live Embryos by Selective Plane Illumination Microscopy , 2004, Science.

[11]  Dean Wilding,et al.  High-speed 2D and 3D fluorescence microscopy of cardiac myocytes. , 2011, Optics express.

[12]  Philipp J. Keller,et al.  Reconstructing embryonic development , 2011, Genesis.

[13]  William E. Ortyn,et al.  Cellular image analysis and imaging by flow cytometry. , 2007, Clinics in laboratory medicine.

[14]  Phil F. Culverhouse,et al.  Automatic image analysis of plankton: future perspectives , 2006 .

[15]  Jan Huisken,et al.  Selective plane illumination microscopy techniques in developmental biology , 2009, Development.

[16]  Farooq Azam,et al.  Thin laser light sheet microscope for microbial oceanography. , 2002, Optics express.

[17]  Laura Bonetta,et al.  Flow cytometry smaller and better , 2005, Nature Methods.