High-throughput label-free image cytometry and image-based classification of live Euglena gracilis.

We demonstrate high-throughput label-free single-cell image cytometry and image-based classification of Euglena gracilis (a microalgal species) under different culture conditions. We perform it with our high-throughput optofluidic image cytometer composed of a time-stretch microscope with 780-nm resolution and 75-Hz line rate, and an inertial-focusing microfluidic device. By analyzing a large number of single-cell images from the image cytometer, we identify differences in morphological and intracellular phenotypes between E. gracilis cell groups and statistically classify them under various culture conditions including nitrogen deficiency for lipid induction. Our method holds promise for real-time evaluation of culture techniques for E. gracilis and possibly other microalgae in a non-invasive manner.

[1]  Ho Cheung Shum,et al.  Optofluidic time-stretch imaging - an emerging tool for high-throughput imaging flow cytometry. , 2016, Lab on a chip.

[2]  Aram J. Chung,et al.  Three dimensional, sheathless, and high-throughput microparticle inertial focusing through geometry-induced secondary flows. , 2013, Small.

[3]  Shizhong Xie,et al.  Time-stretch high-speed microscopic imaging system based on temporally and spectrally shaped amplified spontaneous emission. , 2015, Optics letters.

[4]  Cheng Lei,et al.  Optical time-stretch imaging: Principles and applications , 2016 .

[5]  B. Bouma,et al.  Three-dimensional miniature endoscopy , 2006, Nature.

[6]  Shizhong Xie,et al.  Multiwavelength time-stretch imaging system. , 2014, Optics letters.

[7]  Flow cytometry using spectrally encoded confocal microscopy. , 2010, Optics letters.

[8]  R. Sims,et al.  Production and harvesting of microalgae for wastewater treatment, biofuels, and bioproducts. , 2011, Biotechnology advances.

[9]  J. Myers,et al.  Growth and photosynthetic characteristics of euglena gracilis , 2004, Archiv für Mikrobiologie.

[10]  K. Goda,et al.  Dispersive Fourier transformation for fast continuous single-shot measurements , 2013, Nature Photonics.

[11]  B. Jalali,et al.  Serial time-encoded amplified imaging for real-time observation of fast dynamic phenomena , 2009, Nature.

[12]  Y Summar,et al.  UV-induced changes in phytoplankton cells and its effects on grazers , 1997 .

[13]  Cheng Lei,et al.  High-throughput optofluidic particle profiling with morphological and chemical specificity. , 2015, Optics letters.

[14]  S. Mayfield,et al.  Exploiting diversity and synthetic biology for the production of algal biofuels , 2012, Nature.

[15]  Chi Zhang,et al.  High-performance multi-megahertz optical coherence tomography based on amplified optical time-stretch. , 2015, Biomedical optics express.

[16]  G. Whitesides,et al.  Soft Lithography. , 1998, Angewandte Chemie.

[17]  Ata Mahjoubfar,et al.  Optical Data Compression in Time Stretch Imaging , 2015, PloS one.

[18]  郭强,et al.  Fast time-lens-based line-scan single-pixel camera with multi-wavelength source , 2015 .