Lensfree Fluorescence Microscopy
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Lensfree Fluorescence Microscopy

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Optical microscopes enable us to see micron sized objects with high sensitivity and resolution. However, the field-of-view (FOV) of conventional lens-based imaging systems is typically limited to less ...

Optical microscopes enable us to see micron sized objects with high sensitivity and resolution. However, the field-of-view (FOV) of conventional lens-based imaging systems is typically limited to less than a few mm2 which in general requires mechanical scanning to image e.g., the entire area of large lab-on-a-chip platforms. To address this limitation, here we demonstrate an on-chip microscopy platform that can image fluorescently labeled cells or small model animals (e.g. C. Elegans) over a wide FOV of >0.6-8 cm2, achieving <4μm><4μm>0.6-8 cm2.This compact and wide-field on-chip fluorescent imaging platform, with rapid digital decoders behind it, could be rather valuable for high-throughput cytometry, rare-cell analysis and microarray research.

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Lensfree Fluorescence Microscopy Lensfree Fluorescence Microscopy Presentation Transcript

  • Lensfree Fluorescence Microscopy  Ahmet  F.  Coskun*,  Ting-­‐Wei  Su,  Ikbal  Sencan,  Aydogan  Ozcan     Electrical  Engineering  Department,   University  of  California,  Los  Angeles   hHp://innovate.ee.ucla.edu/   *afc@ucla.edu     2011 1  
  •        Lens-­‐based  fluorescence  imaging  systems  have  been  used     for  high-­‐throughput  screening  applica9ons   Circula0ng  Tumor  Cell    Analysis   Microarray  Research   C.  Elegans  Imaging   Ozcan  Research  Group,  Department  of  Electrical  Engineering,  University  of  California  at  Los  Angeles   2  
  • Typical  imaging  FOV  of  such  lens-­‐based  systems  is          not  yet  compa9ble  with  the  scale  of  lab-­‐on-­‐a-­‐chip  pla?orms   … requires scanning to image the entire lab-on-a-chip platformsTowards this end, there is a need to simultaneously image an ultra wide field-of-view for high-throughput screening applications Ozcan  Research  Group,  Department  of  Electrical  Engineering,  University  of  California  at  Los  Angeles   3  
  • Ultra  high-­‐throughput  screening  pla?orm  (LUCAS)   Lens-free On-chip Imaging PlatformFor Fluorescent and Holographic Microscopy of Cells or Small model animals Over >18 cm2 Field-of-view All in parallel within seconds 20X Lensless Ultra wide-field Cell monitoring Array platform based on Shadow imaging (LUCAS) Ozcan  Research  Group,  Department  of  Electrical  Engineering,  University  of  California  at  Los  Angeles   4  
  • Lensfree  Fluorescent  and  Holographic  imaging  pla?orm   Incoherent Illumination for Lensless Holography Prism Micro-fluidic Chip TIR Absorption Filter (<100µm) Surface CCD/CMOS Emission Ozcan  Research  Group,  Department  of  Electrical  Engineering,  University  of  California  at  Los  Angeles   5  
  • LUCAS  pla?orms  are   compact  and  easily  integrated  with  microfluidic  chips   Volume 136 | Number 17 | 2011 Analyst Interdisciplinary detection science www.rsc.org/analyst Volume 136 | Number 17 | 7 September 2011 | Pages 3393–3572 Analyst Themed Issue on Emerging InvestigatorsShowcasing research from the Bio-Photonics As featured in:Laboratory at UCLA led by Professor Aydogan Ozcan,a faculty member at the Electrical Engineering andBioengineering Departments. Ozcan  Research  Group,  Department  of  Electrical  Engineering,  University  of  California  at  Los  Angeles  Title: Wide- eld lensless uorescent microscopy using a tapered 6  
  • The  key  principles  of  on-­‐chip  microscopy  system   400  µm   400  µm   400  µm  Lensfree  Fluorescent     The  use  of  planar     With  Digital  Decoders   Encoder     + Op0cal  components   + (  Compressive  sensing     (  Faceplate  and  Taper)     based  Decoding)   On  –  chip  Lensless  Fluorescent  Microscopy     Over  >  0.6-­‐  8  cm2   With  >  4  µm  spa0al  resolu0on   Ozcan  Research  Group,  Department  of  Electrical  Engineering,  University  of  California  at  Los  Angeles   7  
  • Planar  fiber  array  components  (Faceplate)     are  used  to  engineer  PSF  Faceplate   Prism Micro-fluidic Chip TIR Faceplate (>3mm) Surface Absorption Filter (<100µm) Emission CCD/CMOS 8   Ozcan  Research  Group,  Department  of  Electrical  Engineering,  University  of  California  at  Los  Angeles  
  • Planar  fiber  array  components  (Faceplate)     are  used  to  engineer  PSF  Faceplate   400  µm   400  µm   w/o  faceplate   With  faceplate  
  • Planar  fiber  array  components  (Tapered  Faceplate)     are  used  to  introduce  magnifica9on  Tapered  Faceplate   2.2 µm Prism Micro-fluidic Chip TIR 4.9 µm Surface TAPER (>15mm) Absorption Filter 10   CCD/CMOS Emission Ozcan  Research  Group,  Department  of  Electrical  Engineering,  University  of  California  at  Los  Angeles  
  • Magnifica9on  is  introduced  to  increase  the  resolu9on   Demagnification Magnification20  µm   100  µm   d d d/2.3 d * 2.3 Ozcan  Research  Group,  Department  of  Electrical  Engineering,  University  of  California  at  Los  Angeles   11  
  • The  key  principles  of  on-­‐chip  microscopy  system   400  µm   400  µm   400  µm  Lensfree  Fluorescent     The  use  of  planar     With  Digital  Decoders   Encoder     + Op0cal  components   + (  Compressive  sensing     (  Faceplate  and  Taper)     based  Decoding)   On  –  chip  Lensless  Fluorescent  Microscopy     Over  >  0.6-­‐  8  cm2   With  >  4  µm  spa0al  resolu0on   Ozcan  Research  Group,  Department  of  Electrical  Engineering,  University  of  California  at  Los  Angeles   12  
  • Lensfree  fluorescent  encoding  and  decoding  system  Ozcan  Research  Group,  Department  of  Electrical  Engineering,  University  of  California  at  Los  Angeles   13  
  • An  Emerging  theory:  Compressive  Sensing/Sampling   Compressive  Sampling:  A  mathema0cal  framework  enables   unique  recovery  of  a  signal  from  much  fewer  samples  than   normally  required  according  to  the  sampling  theorem   200 µm 200 µm 200 µm 200 µm Developed  by  a  group  of  Mathema0cians;  Boyd  (  Stanford),  Candes  (  Caltech),  Donoho(  Stanford),  Tao(  UCLA)   Ozcan  Research  Group,  Department  of  Electrical  Engineering,  University  of  California  at  Los  Angeles   14  
  •                            Fluorescent  Imaging  can  also  benefit  from  CS  for  reconstruc9on   Lensfree fluorescent images are already SPARSE à 400  µm   Measured Raw Fluorescent Image Unknown Object Distribution 2D Measured Incoherent PSF Regularization ParameterSeung-Jean Kim; Koh, K.; Lustig, M.; Boyd, S.; Gorinevsky, D.; , "An Interior-Point Method for Large-Scale l1-Regularized LeastSquares," Selected Topics in Signal Processing, IEEE Journal of , vol.1, no.4, pp.606-617, Dec. 2007 Ozcan  Research  Group,  Department  of  Electrical  Engineering,  University  of  California  at  Los  Angeles   15  
  • 2  µm  fluorescent  micro-­‐objects     are  used  to  characterize  resolu9on  Ozcan  Research  Group,  Department  of  Electrical  Engineering,  University  of  California  at  Los  Angeles   16  
  • <  4  µm  spa9al  resolu9on  is  verified     By  resolving  2  µm  bead  pairs     < 4 µm Lateral Resolution Note that the pixel size of the sensor is 9 µm (Resolution is NOT limited by pixel size)Ozcan  Research  Group,  Department  of  Electrical  Engineering,  University  of  California  at  Los  Angeles   17  
  • White  blood  cells  are  imaged     over  >  8cm2  for  on-­‐chip  cytometry   JOVE 2011 Required Computation time:Using CPU Computer ~ 10 min for entire FOV ~ 1 sec for zoomed FOV Can be done faster using GPUs and CUDAs Ozcan  Research  Group,  Department  of  Electrical  Engineering,  University  of  California  at  Los  Angeles   18  
  •                      Imaging  of  Giardia  Muris  Cysts  for  Water  Quality  Monitoring   (a) Lensfree Raw Image 200  µm   Analyst 2011 (b) CS Decoded Image (c) 10X Microscope Image 50  µm   50  µm   Ozcan  Research  Group,  Department  of  Electrical  Engineering,  University  of  California  at  Los  Angeles   19  
  • C.Elegans  imaging  for  On-­‐chip  Drug  Screening   PLoS ONE 2011Ozcan  Research  Group,  Department  of  Electrical  Engineering,  University  of  California  at  Los  Angeles   20  
  • MCF7  Cancer  Cell  Imaging  for  Rare  Cell  Research   Biosensor BC 2011Ozcan  Research  Group,  Department  of  Electrical  Engineering,  University  of  California  at  Los  Angeles   21  
  • 3D  Imaging  of  ver9cally  separated  micro-­‐channels   Recorded Lensfree Raw CS Decoder First Layer Result CS Decoder Second Layer(a)   (b)   Image Z+Δz Result Z Lensfree Raw Δz Layer 2 Layer 1 Image Z Lensfree Raw Image (Faceplate) Lensfree Raw Image 200 µm Optics Express 2011 Ozcan  Research  Group,  Department  of  Electrical  Engineering,  University  of  California  at  Los  Angeles   22  
  •                              Lensfree  fluorescent  imaging                    for  high-­‐throughput  on-­‐chip  analysis  C.Elegans Imaging Rare Cell Screening Microarray Analysis Ozcan  Research  Group,  Department  of  Electrical  Engineering,  University  of  California  at  Los  Angeles   23  
  •                          References  (1)  Ahmet  F.  Coskun,  Ting-­‐Wei  Su  and  Aydogan  Ozcan,  "Wide  field-­‐of-­‐view  lens-­‐free  fluorescent  imaging  on  a  chip,"    Lab  Chip,  10,  824  (2010)    (2)  Ahmet  F.  Coskun*,  Ikbal  Sencan*,  Ting-­‐Wei  Su*,  and  Aydogan  Ozcan,  "Lensless  wide-­‐field  fluorescent  imaging  on  a  chip  using  compressive  decoding  of  sparse  objects,"  Opt.  Express  18,  10510-­‐10523  (2010)    (3)  Ahmet  F.  Coskun,  Ting-­‐Wei  Su,  Ikbal  Sencan,  and  Aydogan  Ozcan,  "Lensfree  fluorescent  on-­‐chip  imaging  over  an  ultra-­‐wide  field-­‐of-­‐view  using  compressive  sampling,"Opacs  &  Photonics  News  21(12),  27-­‐27  (2010)    (4)  Ahmet  F.  Coskun*,  Ikbal  Sencan*,  Ting-­‐Wei  Su*,  and  Aydogan  Ozcan,  "Lensfree  Fluorescent  On-­‐Chip  Imaging  of  Transgenic  Caenorhabdias  elegans  Over  an  Ultra-­‐Wide  Field-­‐of-­‐View,"  PLoS  ONE  (2011)    (5)  Ahmet  F.  Coskun*,  Ikbal  Sencan*,  Ting-­‐Wei  Su*,  and  Aydogan  Ozcan,  "  Wide-­‐field  Lensless  Fluorescent  Microscopy  Using  a  Tapered  Fiber-­‐opac  Faceplate  On  a  Chip,"  Analyst  (2011)  (6)  Ahmet  F.  Coskun,  Ting-­‐Wei  Su,    Ikbal  Sencan,  and  Aydogan  Ozcan,  ”  Lensless  Fluorescent  Microscopy  On  a  Chip,"  JOVE  (2011)     *Equal  Contribuaon   Ozcan  Research  Group,  Department  of  Electrical  Engineering,  University  of  California  at  Los  Angeles   24  
  • Acknowledgements  –  Ozcan  Research  Group  @  UCLA   Research  Website  for  more  informa9on:   hbp://innovate.ee.ucla.edu   Ozcan  Research  Group,  Department  of  Electrical  Engineering,  University  of  California  at  Los  Angeles   25