Holographic Video FlyThroughs of Osteogenic Tumor Spheroids

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Holographic Video Fly-Throughs of Osteogenic
Tumor Spheroids
Ping Yu Department of Physics, Purdue
University, USA 
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• Outline
• Introduction.
• A new imaging technique holographic Optical
  Coherence Imaging (OCI).
• Heterogeneities from Necrosis and calcification.
• Shimmering Holograms and Cellular Movement.
• Conclusion and future work.

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D. D. Nolte M. Mustata L. Peng Biophotonic
Laboratory Department of Physics Purdue
University, USA 
P.M.W. French C. Dunsby Y. Gu Photonics
Group Imperial College, UK
J. J. Turek Department of Basic Medical
Sciences School of Veterinary Medicine Purdue
University, USA M. R. Melloch School of
Electrical and Computer Engineering Purdue
University, USA
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• A brief history of biomedical imaging
• Computed tomography (CT). Allan Cormack and
  Godfrey Hounsfield 1971. 0.3mm
• Magnetic resonance imaging (MRI) 1978-1985. 1 mm
• Ultrasound imaging and PET.
• Optical methods.

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Low coherent light source
Imaging system
Sample
PRQW

• Optical coherence imaging (OCI) is a full-frame
  variant of optical coherence tomography (OCT).
• The coherent image spatially modulates a high
  sensitivity and high speed dynamic holographic
  film composed of a photorefractive quantum well
  (PRQW).

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Degenerate Four-wave mixing
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Experimental Set-up of OCI
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Lateral resolution 10 ?m
Depth resolution 30 ?m
(a)
(b)
Holograms of USAF test-chart (a) in lateral
plane, and (b) cross section in axial plane.
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Tumor spheroids
Tumor spheroids
Tumor cells
Growth conditions

• Tumor spheroids size several tens ?m to several
  hundreds ?m.
• Tumor cell size 10 ?m
• Refractive index difference between cell nuclei,
  membrane and environment

?n 0.04
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Differential interference contrast image of a rat
osteogenic tumor spheroid.
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610 ?m
Input laser
Coherent gate scan direction
Holographic Fly-through of tumor spheroids.
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Input laser
610 ?m
3D view of tumor spheroids from OCI data.
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Optical micrograms of thin section of tumor
spheroid.
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B
A
(A) (B)
Transmission electron micrograph of outer living
spheroid shell (A) and inner core (B) with areas
of necrosis and mineralization.
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2
3
4
1
7
8
6
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Holograms from OCI of tumor spheroids.
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Input laser
255
610 ?m
255
0
Cross-section image of the tumor spheroid.
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Average intensity as a function of radius from
different samples.
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800 ?m
A
C
B
y
x
Three tumor spheroids samples attach together.
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A
x
depth
B
z (depth)
Cross-section at y114 from three tumor
spheroids.
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A
x
depth
B
z depth
Cross-section at y127 from three tumor
spheroids.
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B
y
depth
C
C
z (depth)
Cross-section at x68 from three tumor spheroids.
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Tumor spheroids in hypotonic, isotonic,
hypertonic environments.
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Multicellular spheroids as a realistic model to
study tumor response to agents and radiation, or
tumor cells response to drug. Find a way to
locate area of necrotic cells and apoptotic
cells.
Kenneth M. Yamada and Katherine C., Nature 419,
790 - 791 (2002)
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Input laser
Coherent gate stop inside the specimen
Fly-through of fresh tumor spheroids.
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Input laser
Coherent gate stop inside the specimen
Fly-through of metabolic poisoned tumor
spheroids. Sodium azide is used as a poison.
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Input laser
Coherent gate stop inside the specimen
Fly-through of crosslinked tumor spheroids.
Glutaraldehyde is used as a poison.
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Fresh tumor showing average, maximum and standard
deviation. The fourth fame is the feature map for
persistent and variable features.
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Metabolically poisoned tumor showing
average,maximum and standard deviation. The
fourth fame is the feature map for persistent and
variable features.
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Crosslinked tumor showing average,maximum and
standard deviation. The fourth fame is the
feature map for persistent and variable features.
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• Conclusion and Future work
• First fly-throughs of tumor spheroids using
  holographic OCI.
• Distribution of necroses and calcifications can
  be analysis using OCI data while the living cells
  are almost invisible.
• The deepest information, according to our data
  sets, can be achieved for about 600 ?m beneath
  the surface of the tissue.
• Holographic OCI provides a powerful tool to get
  real-time structure of biological tissues.
• High resolution OCI is possible in near future.