Interferometric Mic In icroscopy for Detection and Vis - PowerPoint PPT Presentation

Interferometric Mic In icroscopy for Detection and Vis isualization of f Bio iological Nanoparticles M. Selim Ünlü Electrical Engineering, Physics, Biomedical Engineering Graduate Medical Sciences BUNano Photonics Center CLEO 2018 selim@bu.edu www.bu.edu/OCN 1/32

Outline • Motivation – Biological Nanoparticles everywhere • Problem definition – contrast and size • Detection vs. visualization • Interferometric Reflectance Imaging Sensor • Biological Nanoparticle Detection and Sizing • Pupil function engineering • Resolution improvement by oblique illumination and reconstruction • Towards 100nm in label-free visible light microscopy • Conclusions and Future CLEO 2018 selim@bu.edu www.bu.edu/OCN 2/32

Extra cellular vesicles, exosomes, and viruses � SEM image of Ebola virion Example cryo-EM images of Viruses are the most abundant species on earth. infectious extracellular vesicle ~10 32 phages in the biosphere (Bullitt Lab – BU MED) ~10 7 viruses on average in a mL of seawater CLEO 2018 selim@bu.edu www.bu.edu/OCN 3/32

Optical microscopy can see small – but … micro.magnet.fsu.edu/primer/ CLEO 2018 selim@bu.edu www.bu.edu/OCN 4/32

Biological Nanoparticle Detection Challenges – size and dielectric contrast E inc E sca Ziegler       p m 3 4 R Signal ~ R 6    0 2 p m Size contrast CLEO 2018 selim@bu.edu www.bu.edu/OCN 5/32

Technologies for bio-nanoparticle characterization Cryo-TEM Nanoparticle Tracking Analysis Fluorescence microscopy • Fantastic resolution • Estimate size of particles based (STED/PALM ) • Low throughput and difficult on Brownian motion • Little/no molecular information Needed: High-throughput methods to measure the size, shape and molecular profile of biological nanoparticles CLEO 2018 selim@bu.edu www.bu.edu/OCN 6/32

2   2   E ref I E E 2 E E sin det ref sca ref sca E sca       Phase Term p m 3 4 R    0 2 p m SiO 2 Si Size Material CLEO 2018 selim@bu.edu www.bu.edu/OCN 7/32

I nterferometric ic R efl flectance I magin ing S ensor (IR (IRIS IS) a hig igh throughput bio iosensin ing pla latform soap film Oxide coated Si Protein microarray chips with 100s to 1,000s of probe spots @ $0.1/cm 2 Ünlü et al, ”STRUCTURED SUBSTRATES FOR OPTICAL SURFACE PROFILING,’ US Patent No: 9599611, 2017 pg/mm 2 sensitivity 1,000s of spots CLEO 2018 selim@bu.edu www.bu.edu/OCN 8/32

Simple Single Particle Detection Rahul Vedula(MD) and George Daaboul, PhD ‘13 CLEO 2018 selim@bu.edu www.bu.edu/OCN 9/32

Exosome detection Anti-CD81 capture probe image acquired before and after incubation with purified HEK293 cells derived exosomes. CLEO 2018 selim@bu.edu www.bu.edu/OCN 10/32

Virus characterization: size determination SEM IRIS Daaboul et al, ACS nano 8 (2014) CLEO 2018 selim@bu.edu www.bu.edu/OCN 11/32

Various viruses CLEO 2018 selim@bu.edu www.bu.edu/OCN 12/32

In-liquid detection to simplify the assay 13 CLEO 2018 selim@bu.edu www.bu.edu/OCN 13/32

Virus detection for diagnostic applications Highly-sensitive virus detection directly from blood serum Scherr et al, ACS Nano 10 (2016) and Scherr et al, Lab on a Chip (20 17) CLEO 2018 selim@bu.edu www.bu.edu/OCN 14/32 (2017)

Passive Cartridge - Simple Workflow 1. Remove cartridge from package just prior to use 2. 100 uL of sample is pipetted into the bottom of the reservoir (*care needs to be taken not to introduce bubbles) 3. Luer cap (sealed with adhesive strip) is screwed down finger tight 4. When liquid reaches the pad, the luer cap is vented (adhesive strip removed) 5. Cartridge is placed in the instrument to begin acquiring data ‘17 ‘15 CLEO 2018 selim@bu.edu www.bu.edu/OCN 15/32

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Interferometric fringes – defocus profile Changing the focus position changes the path length to the detector differently for reference reflection and ‘17 scattered light ‘17 D. Sevenler et al, "Quantitative interferometric reflectance imaging for the detection and measurement of biological nanoparticles," Biomedical Optics Express , 2017 O. Avci, et al., "Physical Modeling of Interference Enhanced Imaging and Characterization of Single Nanoparticles," Optics Express , 2016 O. Avci, et al. "Pupil function engineering for enhanced nanoparticle visibility in wide-field interferometric microscopy," Optica 2017 CLEO 2018 selim@bu.edu www.bu.edu/OCN 17/32

‘17 CLEO 2018 selim@bu.edu www.bu.edu/OCN 18/32

Outline – Going Beyond Detection and Sizing • Motivation – Biological Nanoparticles everywhere • Problem definition – contrast and size • Detection vs. visualization • Interferometric Reflectance Imaging Sensor • Biological Nanoparticle Detection and Sizing • Pupil function engineering • Resolution improvement by oblique illumination and reconstruction • Towards 100nm in label-free visible light microscopy • Conclusions and Future CLEO 2018 selim@bu.edu www.bu.edu/OCN 19/32

‘17 Overall of 10X enhancement CLEO 2018 selim@bu.edu www.bu.edu/OCN 20/32

Collection Path – Apodization and Reference Attenuation CLEO 2018 selim@bu.edu www.bu.edu/OCN 21/32

Silica particles defocus curve ~5X enhancement (3 (3% → 15%) ) a)฀full-NA฀illumination฀at฀ λ =530฀nm,฀b)฀apodized฀illumination฀at฀ λ =530฀nm,฀c)฀apodized฀illumination฀at฀ λ =460฀nm,฀d)฀apodized฀ illumination฀with฀amplitude฀filter฀in฀the฀collection฀path฀at฀ λ =460฀nm. CLEO 2018 selim@bu.edu www.bu.edu/OCN 22/32

A DVANCED W IDE - FIELD I NTERFEROMETRIC M ICROSCOPY F OR N ANOPARTICLE S ENSING A ND C HARACTERIZATION Reconstruction in In Interference Microscopy imaging system object observation in out (J. Trueb*, O. Avci* et al., IEEE JSTQE, 2016) ? observation noise system response object convolution matrix CLEO 2018 selim@bu.edu www.bu.edu/OCN 23/32

฀ ฀ ฀NATURE฀PHOTONICS฀|฀VOL฀8฀|฀MAY฀2014฀|฀฀ CLEO 2018 selim@bu.edu www.bu.edu/OCN 24/32

A DVANCED O PTICAL S CHEMES IN W IDE - FIELD I NTERFEROMETRIC M ICROSCOPY F OR E NHANCED N ANOPARTICLE S ENSING A ND C HARACTERIZATION Super-resolution in wide-field interferometric microscopy • Enhancing low-index nanoparticle resolution via reconstruction schemes Asymmetric illumination based reconstruction for super resolution (with Lei Tian) Fourier transforms of the transfer functions (H) for each asymmetric illumination configuration Left Right Bottom Top 10/2/2018 CLEO 2018 selim@bu.edu www.bu.edu/OCN 25/32

Experimental Results reconstruction raw Sample – E-beam fabricated 50x/0.8NA 525nm SEM Sketch 50x/0.8NA 525nm 300 nm 100x/0.9NA 525nm SEM 80 nm oxide Si 10/2/2018 CLEO 2018 selim@bu.edu www.bu.edu/OCN 26/32

150 nm separation, 0.9 NA, 𝝻 = 420nm FWHM ~ 130nm < ( 𝝻 / 3) CLEO 2018 selim@bu.edu www.bu.edu/OCN 27/32

Elongated polystyrene rods Samir Mitragotri (Harvard) Full NA Reconstructed image CLEO 2018 selim@bu.edu www.bu.edu/OCN 28/32

Commercialization Nanoview ZOIRAY CLEO 2018 selim@bu.edu www.bu.edu/OCN 29/32

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CONCLUSIONS & FUTURE • Optical interference is a very powerful sensing technique. • Multi-disciplinary innovation � • Single biological nanoparticle detection / counting / size and shape discrimination / visualization • Goals: Down to r=20nm Biological nanoparticle detection in liquid • Lateral resolution of ~100nm without labeling CLEO 2018 selim@bu.edu www.bu.edu/OCN 31/32