High resolution light-sheet based high-throughput imaging cytometry system enables visualization of intra-cellular organelles

Visualization of intracellular organelles is achieved using a newly developed high throughput imaging cytometry system. This system interrogates the microfluidic channel using a sheet of light rather than the existing point-based scanning techniques. The advantages of the developed system are many, including, single-shot scanning of specimens flowing through the microfluidic channel at flow rate ranging from micro- to nano- lit./min. Moreover, this opens-up in-vivo imaging of sub-cellular structures and simultaneous cell counting in an imaging cytometry system. We recorded a maximum count of 2400 cells/min at a flow-rate of 700 nl/min, and simultaneous visualization of fluorescently-labeled mitochondrial network in HeLa cells during flow. The developed imaging cytometry system may find immediate application in biotechnology, fluorescence microscopy and nano-medicine.

[1]  A. Sen,et al.  Microfluidic System for Rapid Enumeration and Detection of Microparticles , 2012 .

[2]  Y. K. Cheung,et al.  1 Supplementary Information for : Microfluidics-based diagnostics of infectious diseases in the developing world , 2011 .

[3]  Aydogan Ozcan,et al.  On-Chip Cytometry using Plasmonic Nanoparticle Enhanced Lensfree Holography , 2013, Scientific Reports.

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

[5]  P. Mondal,et al.  Generation of extended light-sheets for single and multi-photon fluorescence microscopy , 2013 .

[6]  Josef Kittler,et al.  A survey of the hough transform , 1988, Comput. Vis. Graph. Image Process..

[7]  Donald Wlodkowic,et al.  Microfluidic single-cell array cytometry for the analysis of tumor apoptosis. , 2009, Analytical chemistry.

[8]  L L Wheeless,et al.  Imaging in flow. , 1979, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[9]  Aaron M. Streets,et al.  Ostwald ripening of clusters during protein crystallization. , 2010, Physical review letters.

[10]  D H Burns,et al.  Orthogonal‐plane fluorescence optical sectioning: Three‐dimensional imaging of macroscopic biological specimens , 1993, Journal of microscopy.

[11]  T. Holy,et al.  Fast Three-Dimensional Fluorescence Imaging of Activity in Neural Populations by Objective-Coupled Planar Illumination Microscopy , 2008, Neuron.

[12]  Yanhui Zhao,et al.  An integrated, multiparametric flow cytometry chip using "microfluidic drifting" based three-dimensional hydrodynamic focusing. , 2012, Biomicrofluidics.

[13]  Daniel J. Ehrlich,et al.  A Parallel Microfluidic Flow Cytometer for High Content Screening , 2011, Nature Methods.

[14]  R. Zengerle,et al.  Microfluidic lab-on-a-chip platforms: requirements, characteristics and applications. , 2010, Chemical Society reviews.

[15]  S. Quake,et al.  A microfabricated fluorescence-activated cell sorter , 1999, Nature Biotechnology.

[16]  A. Diaspro,et al.  Live-cell 3D super-resolution imaging in thick biological samples , 2011, Nature Methods.

[17]  S. Quake,et al.  A Systems Approach to Measuring the Binding Energy Landscapes of Transcription Factors , 2007, Science.

[18]  Jeff Squier,et al.  Microfluidic cell counter with embedded optical fibers fabricated by femtosecond laser ablation and anodic bonding. , 2009, Optics express.

[19]  G Valet,et al.  Fast imaging in flow: a means of combining flow-cytometry and image analysis. , 1979, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[20]  Kenji Yasuda,et al.  Fully Automated On-Chip Imaging Flow Cytometry System with Disposable Contamination-Free Plastic Re-Cultivation Chip , 2011, International journal of molecular sciences.

[21]  Hongying Zhu,et al.  Optofluidic Tomography on a Chip. , 2011, Applied physics letters.

[22]  T. Liao,et al.  A cell counting/sorting system incorporated with a microfabricated flow cytometer chip , 2006 .

[23]  Philipp J. Keller,et al.  Fast, high-contrast imaging of animal development with scanned light sheet–based structured-illumination microscopy , 2010, Nature Methods.

[24]  J. Friend,et al.  Fabrication of microfluidic devices using polydimethylsiloxane. , 2010, Biomicrofluidics.

[25]  Guiding the migration of adherent cells by using optical micropatterns , 2013 .

[26]  A. Schierloh,et al.  Ultramicroscopy: three-dimensional visualization of neuronal networks in the whole mouse brain , 2007, Nature Methods.

[27]  Yu Sa,et al.  Study of low speed flow cytometry for diffraction imaging with different chamber and nozzle designs , 2013, Cytometry. Part A : the journal of the International Society for Analytical Cytology.

[28]  Daniel A Fletcher,et al.  Analyzing cell mechanics in hematologic diseases with microfluidic biophysical flow cytometry. , 2008, Lab on a chip.

[29]  Valery V Tuchin,et al.  In vivo multispectral, multiparameter, photoacoustic lymph flow cytometry with natural cell focusing, label‐free detection and multicolor nanoparticle probes , 2008, Cytometry. Part A : the journal of the International Society for Analytical Cytology.