Serial Time-Encoded Amplified Microscopy (STEAM) toward High-

Throughput Identification and Enumeration of Rare CellsKeisuke Goda, Ali Motafakker-Fard, Kevin K. Tsia, and Bahram Jalali

Photonics Laboratory, Department of Electrical Engineering University of California, Los Angeles

CLEO Europe, Munich, Germany

June 14, 2009

Neural activity dynamics

Measuring Ca2+

transients from neurons

NAD(P)H waves traveling along a neutrophils long axis

Chemical dynamics in living cells

1 2

3 4

Microfluidics and high-throughput screening (HTS)

High Demand for High-Speed Imaging

Speed Limitations in Conventional Imaging

Traditional electronic image sensors are slow due to 1. Mechanical scanning (~10 kHz)2. Charge download time in CCD/CMOS cameras (~1 kHz)

CW LaserLaser

Scanner

Download

Data Rate = ~10 MHz

More importantly, there is the fundamental trade-off: Sensitivity vs.

Speed

Spectrally Encoded Imaging

1.

H. O. Bartelt, Opt. Commun. 27, 365 (1978)

image transmission using a single-mode fiber2.

G. J. Tearney, M. Shishkov, and B. E. Bouma, Opt. Lett. 27, 412 (2002)

miniature endoscopy3.

K. Shi, P. Li, S. Yin, and Z. Liu, Opt. Express. 12, 2096 (2004)

confocal microscopy

Lens

Diffraction

Grating

Sample

Broadband Optical Pulses

Probe Spectrum Reflected Spectrum Reconstructed Image

time

Amplified dispersive Fourier Transform

time

wavelength

time

wavelength wavelength

time

dispersionsample

Amplified dispersive Fourier transform

= an optical process that maps the spectrum of an optical pulse into a time-domain waveform using group-velocity dispersion and amplifies it simultaneously

1.

D. R. Solli, J. Chou, and B. Jalali, Nature Photon. 2, 48 (2008)2.

K. Goda, K. K. Tsia, and B. Jalali, Nature 458, 1145 (2009)

Real-time pulse-by-pulse spectrum analysis

http://images.google.com/imgres?imgurl=http://shop.c-illuminations.com/images/Spool.jpg&imgrefurl=http://shop.c-illuminations.com/index.php%3FcPath%3D27%26language%3Den%26osCsid%3Dcc2e0471cc8022ce142d5d5ffad26504&h=720&w=720&sz=19&hl=en&start=6&tbnid=YwUE470vDMvXAM:&tbnh=140&tbnw=140&prev=/images%3Fq%3Dfiber%2Boptic%2Bspool%26gbv%3D2%26ndsp%3D20%26hl%3Den%26sa%3DN

1.3 ns/nm

c

= 1565 nm

= 17 nm

Rep. Rate = 25 MHz

Serial Time-Encoded Amplified Microscopy

1DUltrafast Displacement Sensing

Ultrafast Barcode Reading

No need for mechanical scanning

No need for a CCD/CMOS camera

Frame Rate = Pulse Rep. Rate

K. Goda, K. K. Tsia, and B. Jalali, Appl. Phys. Lett. 93, 131109 (2008)

Serial Time-Encoded Amplified Microscopy

2D

Amplification in the dispersive fiber overcomes the fundamental trade-off

in imaging between sensitivity and speed.

Frame Rate = 6.1 MHz, Shutter Speed = 440 ps, Optical Image Gain = 25 dB

K. Goda, K. K. Tsia, and B. Jalali, Nature 458, 1145 (2009)

Serial Time-Encoded Amplified Microscopy

2D

K. Goda, K. K. Tsia, and B. Jalali, Nature 458, 1145 (2009)

Real-Time Observation of Fast Dynamic Events

Observation of Laser Ablation (Phase Explosion)

Ablation Laser Wavelength = 2.8 m Pulse Width = 5 ns Peak Power = ~ 1 MW

Real-Time Observation of Fast Dynamic Events

Ultrafast Microfluidic Flow of Particles

K. Goda, K. K. Tsia, and B. Jalali, Nature 458, 1145 (2009)

Particle Material = MetalParticle Size = 10

30 mFlow Speed = 2.4 m/s

Combined with Flow Cytometry

Fluidic system

LaserDe

tect

ors

OpticalFilters

Detector

Circulating Tumor Cells

Tumor

Need for identification and enumeration of rare cells in blood (e.g., circulating tumor cells)

STEAM enables ultrahigh-

throughput morphological characterization of these cells

Throughput = ~1 million cells/s

Summary & Future Work1.

Serial time-encoded amplified microscopy (STEAM) demonstrated

1D

Ultrafast barcode reading and displacement sensing

K. Goda, K. K. Tsia, and B. Jalali, Appl. Phys. Lett. 93, 131109 (2008)

2D

Continuous real-time observation of fast dynamical phenomena

K. Goda, K. K. Tsia, and B. Jalali, Nature 458, 1145 (2009)

2.

Future work

3D

High-throughput identification and enumeration of rare cells based on

3D morphology (to be published)

Combination with fluorescence microscopy

Slide Number 1Slide Number 2Speed Limitations in Conventional ImagingSpectrally Encoded ImagingAmplified dispersive Fourier TransformSerial Time-Encoded Amplified Microscopy 1DSerial Time-Encoded Amplified Microscopy 2DSerial Time-Encoded Amplified Microscopy 2DReal-Time Observation of Fast Dynamic EventsReal-Time Observation of Fast Dynamic EventsCombined with Flow CytometrySummary & Future Work