Wavefront compensation for deep tissue optical microscopy Full - - PowerPoint PPT Presentation

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Wavefront compensation for deep tissue optical microscopy Full correction System correction 60 mW 300 mW vs. 5 m 5 m Depth = 800 micron Meng Cui Tang et.al. Proceedings of the National Academy of Sciences 109, 8434 8439 (2012). HHMI

SLIDE 1

Meng Cui HHMI Janelia Farm Research Campus

Wavefront compensation for deep tissue optical microscopy

5 μm

Full correction 60 mW

5 μm

System correction 300 mW

vs.

Depth = 800 micron

Tang et.al. Proceedings of the National Academy of Sciences 109, 8434‐8439 (2012).

SLIDE 2

Limitations of optical microscopy

Biological tissues are rarely transparent Why optics? Why not sound, x‐ray? Molecular contrast, resolution, and sensitivity

http://beautifulafricanwildanimalspets.blogspot.com/2012/02/jellyfish.html

SLIDE 3

Aberration and scattering

http://en.wikipedia.org/wiki/Mirage

Same physical effect: Wavefront distortion caused by inhomogeneous refractive index

http://www.soundbuzz360.com/cloud/

SLIDE 4

The principle of Optical phase conjugation (OPC) works for both aberration and scattering

Direct focusing without wavefront control What if there is a guide star? OPC

SLIDE 5

Multidither coherent optical adaptive techniques (COAT)

W. B. Bridges, et al. Applied Optics, Vol. 13, Issue 2, pp. 291‐300 (1974)

SLIDE 6

Focusing light through highly scattering medium onto a detector target

Meng Cui, Optics Letters, Vol. 36 Issue 6, pp.870‐872 (2011)

SLIDE 7

Experiment results

Sample: glass diffuser Wavelength: 785 nm Focusing objective: NA 0.5, 20x Observation objective: NA 0.9, 60x

Observed transmission without wavefront correction Observed transmission with wavefront correction Measured wavefront profile

Meng Cui, Optics Letters, Vol. 36 Issue 6, pp.870‐872 (2011)

SLIDE 8

Experiment in action

SLIDE 9

Two‐photon scope

Tang et.al. Proceedings of the National Academy of Sciences 109, 8434‐8439 (2012).

SLIDE 10

Test 1, imaging beads through mouse skull

Fluorescence signal [a.u.]

500 1000 1500

System correction

Fluorescence signal [a.u.]

5000 10000 15000

Full correction

Phase [rad]

IMPACT measured phase 5 m

Tang et.al. Proceedings of the National Academy of Sciences 109, 8434‐8439 (2012).

SLIDE 11

Limited field of view  anisoplanatism

Wavefront correction plane Wavefront correction plane

SLIDE 12

Solution for imaging through intact skull anisoplanatism  isoplanatism

Wavefront correction plane

Image relay

SLIDE 13

Test 2, power dependence

Tang et.al. Proceedings of the National Academy of Sciences 109, 8434‐8439 (2012).

SLIDE 14

Test 3, imaging T cell inside lymph nodes

5 μm

Full correction 60 mW

5 μm

System correction 300 mW

vs.

Depth = 800 micron

Tang et.al. Proceedings of the National Academy of Sciences 109, 8434‐8439 (2012).

SLIDE 15

Test 4, stability in live animal

Tang et.al. Proceedings of the National Academy of Sciences 109, 8434‐8439 (2012).

SLIDE 16

Summary

Purely based on signal intensity measurement
High speed
Aberration + scattering
Simple modification to TP scope

SLIDE 17

High speed + large number of degrees of freedom AO

Meng Cui, Optics Express, Vol. 19, Issue 4, pp. 2989‐2995 (2011)

SLIDE 18

k space modulation based wavefront measurement and correction system

Meng Cui, Optics Express, Vol. 19, Issue 4, pp. 2989‐2995 (2011)

SLIDE 19

Test with glass diffuser

Current speed: 1 ms/degree of freedom Total DAQ time: 400 ms Next generation 1‐10 s/mode

Before wavefront correction After wavefront correction Measured wavefront

Meng Cui, Optics Express, Vol. 19, Issue 4, pp. 2989‐2995 (2011)

SLIDE 20

Acknowledgement

Na Ji, Karel Svoboda, Eric Betzig, Charles Shank, Mats Gustafsson, Philipp Keller, Jianyong Tang, Vijay Iyer, Jinyang Liu, Amy Hu, Kerry Sobieski, Lakshmi Ramasamy, Lin Shao, Raju Tomer, Liang Gao, Thomas Planchon

Open Positions: Postdoc Research Scientist

http://www.janelia.org/campus‐community/janelia‐farm‐campus http://www.janelia.org/campus‐community