StructuredIllumination Quantitative Phase Microscopy

1
Structured-Illumination Quantitative Phase
Microscopy
Sri Rama Prasanna Pavani, Ariel Libertun, Sharon
King, and Carol Cogswell Micro Optical Imaging
Systems Laboratory University of Colorado at
Boulder http//moisl.colorado.edu
2
Phase Imaging - What?

• Transparent objects (phase objects) modulate only
  the phase of light
• Square law detectors do not detect phase
  modulations
• Convert phase modulations into detectable
  intensity modulations

Bright field
3
Convert phase modulations into detectable
intensity modulations.
Phase imaging - How?
Digital Holography
Phase contrast
Diff. Interference Contrast

• Quantitative phase after reconstruction
• Thick phase objects
• Single image
• Vibration sensitive
• Phase wrapping

• Quantitative phase for weak phase objects
• No phase wrapping
• Halo and shading-off
• Only for thin objects

• Quantitative phase after reconstruction
• No phase wrapping
• Polarization sensitive
• Only for thin objects
• Multiple images

4
Our method - Why?

• Quantitative phase imaging

Purpose

• Thick, partially absorbing samples
• Polarization insensitive

Applicability

• Single image
• Non-scanning (wide field)
• Non-Iterative

Speed

• Incoherent source
• Inexpensive

Miscellaneous
5
Our method

• Amplitude mask in the field diaphragm
• Pattern is imaged on the sample
• Phase object distorts the pattern
• Record the distorted pattern
• Analytical formula calculates phase

Vs
0.2 0.1
(mm)
0.4 0.2
0 0.2 0.4
(mm)
(mm)
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Our method 1D
2D dot shift
1D dot shift
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Our method 1D

• Analytically relate deformation to the optical
  path length
• Consider a 1D phase object p(x)
• Ray R from point A, after refraction, appears as
  if it originated from B
• Deformation t(x) is the distance between A and B

Normal
Tangent
n2
p(x)
n1
A
B
t(x)
Pavani et al, Quantitative bright field phase
microscopy, to be sent to Applied Optics
8
Our method 2D
1D deformations
After 1D integrations
Quantitative Phase
2D deformation
Pavani et al, Quantitative bright field phase
microscopy, to be sent to Applied Optics
9
Simulation
X 100
18 9 0
5 0 -5
Calculated Phase
Quadratic phase
50 25 0
50 25 0
200 100
200 100
After 1D integrations
1D deformations
X 100
18 9 0
5 0 -5
0 100 200
0 100 200
Error
8 4 0 -4 -8
(nm)
Peak error is 5 orders less than peak phase
Error
0 100 200
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Experimental Results
X,Y Deformations
Dot shift
Original pattern
3 0 -3
360 180
0 240 480
Deformed pattern
3 0 -4
360 180
16.54
0 240 480
Quantitative phase
40 30 20 10 0
Object Drop of optical cement
360 180
480 240 0
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Experiment - Accuracy
Profilometer Our method
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Conclusion

• Quantitative phase imaging in a brightfield
  microscope.
• Phase is calculated from deformation using an
  analytical formula.
• Inexpensive non-scanning, non-iterative,
  single-image technique.

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Acknowledgements

• Prof. Rafael Piestun
• Prof. Gregory Beylkin
• Vaibhav Khire

CDMOptics PhD Fellowship
National Science Foundation Grant No. 0455408
14
References

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Applications and Future work

• Industrial inspection, biological imaging
• Extracting information from axial deformation
• Extending the depth of field of the system
• Fabrication of an amplitude mask with higher
  spatial resolution

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Our method How?
1 Dimensional analysis
(from geometry)
(Snells law,
)
(Taylor expansion)
C 2 (C2 C1)
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Our method How?
M
2 Dimensional analysis
N
Apply 1D solution along x and y to obtain
and
P2