immuno icg nanocapsules and scanning nonlinear optical endomicroscopy for translational biophotonics...
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![Page 1: Immuno ICG Nanocapsules and Scanning Nonlinear Optical Endomicroscopy for Translational Biophotonics Imaging Xingde Li, PhD (1994-1998 PhD at JF) Department](https://reader030.vdocuments.mx/reader030/viewer/2022032606/56649eb45503460f94bbc242/html5/thumbnails/1.jpg)
Immuno ICG Nanocapsules and Scanning Nonlinear Optical Endomicroscopy
for Translational Biophotonics Imaging
Xingde Li, PhD
(1994-1998 PhD at JF)
Department of Biomedical EngineeringJohns Hopkins University
BC Legacy Workshop, June 4, 2011
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Thank you!
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Joining the Lab: June 1994
Fluorescence Imaging in Turbid Media?
Source
rsDetector
rd
phase shift and demodulation due tofluorophore concentration and lifetime
Point Fluorophore
r
ex em
( r )
1 - i ( r )L (r, ) =
Source Intensity
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Near-infrared Fluorescence Contrast Agents
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In vivo Fluorescence SignalsTumor versus Control
0.5
1
1.5
2
2.5
3
3.5
0
0.5
1
1.5
2
0 5 10 15 20 25 30 35 40
Ratio of Max Fluorescenceand Tumor Size
Ra
tio
(T
um
or
/ C
on
tro
l)
Tu
mo
r Size
(cm 3)
Days After Injection of Tumor Cells
Tumor size
Theory
Ratio (T/N)
0
2
4
6
8
10
0 2 4 6 8
Typical Fluorescence Signal
Flu
ore
sc
en
ce
(A
rb.
Un
its
)
Time (min)
Tumor
Normal
Ratio of max fluorescence signal: tumor / normal ~2.5 foldRatio of initial clearance velocity: tumor / normal ~4 fold.
Theoretical contrast: Nobj/Nbg ~7-fold
S-D separation: 8 mmTumor depth: 2 mm (subcutaneous)
Summer 1994
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Preliminary Clinical Study
Patient with breast tumor palpable, ~2cm deep, 0.8 cm in diameterRatio of max fluorescence signal: tumor / normal ~1.5 foldRatio of initial clearance velocity: tumor / normal ~2.0 fold
0.0
0.5
1.0
1.5
2.0
2.5
0 2 4 6 8 10 12
Patient (M84)
Flu
ore
scen
ce
(Arb
. U
nit
s)
Time (min)
Tumor
Normal
Dosage: 0.4 mg / Kg BW( limit ~2.0 mg / Kg BW)
Summer 1994
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Fluorescent Polymeric Micelles
Encapsulate ICG into its hydrophobic core.Enhancing fluorescence emission and stabilityPotentially bioconjugatible, enhanced permeation and retention (EPR) FAD approval?
V. Rodriguez, S. Henry, A. Hoffman, P. Stayton, X. D. Li, and S. H. Pun, Journal of Biomedical Optics (2008)
(pluronic micelles)
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ICG in Chloroform
Stirring 3h Evaporation
Polymer (Aqueous Solution)
OH OH
OHOH
ICG Encapsulation by Pluronic Micelle
(Emulsification and Evaporation)
~30 nm in diameter
ATM Image of ICG Nanocapsules
Size is small, suitable for in vivo tumor delivery.
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ICG Fluorescence Stability of in Micelles
T. H. Kim et al., Pharmaceutical Research 27 (9), 1900-1913 (2010).
Excellent ICG fluorescence stability micellar nanocapsules.
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Tumor
mA
b-m
icel
le/IC
G
Tumor Tumor
Strong ICG fluorescence from targeted tumor with immunomicelles.
In vivo Tumor Imaging with ICG-Immunomicelles
(3 days post injection of ICG micelles with an ICG dose of 0.5 mg/Kg )
XENOGEN IVIS System Tomography System
Tumor
Mic
elle
/ICG Tumor Tumor
nM
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In-tumor ICG Concentration vs Time (Initial Pharmacokinetics Study)
3.0 3.5 4.0 4.5 5.0
4
8
12
16
20
24
Dye
Co
nce
ntr
atio
n (
nM
)
Time (Days)
Immuno-Micelle/ICG Micelle/ICG
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Scanning Nonlinear Endomicroscopy:Enabling in vivo Two-photon Fluorescence (TPF)& Second Harmonic Generation (SHG) Imaging
About Graduation
Chatting with BC about what to do next: Metabolic/Functional Imaging, in vivo
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Two-photon Fluorescence Imaging
Mucosal mast cells: Green: plasma membraneBlue: nucleiRed: mitochondria
www.biorad.com
Nature Biotechnology 21, 1369 - 1377 (2003)
Single-photon Flr Two-photon Flr
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Distal End
Motivation: Nonlinear Optical Microscope Endomicroscope
Accessory Port(2.8 mm)
TPF Endomicroscope
(distal end)
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Photo of Scanning TFP Endomicroscope
Distal End
Report in Biophotonics International, June 2006.M. T. Myaing, D. J. MacDonald, and X. D. Li, Optics Letters 31(8):1076-1078 (2006).
2.4 mm DiameterStandard Endoscope
Endomicroscope
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Visualization of Histopathology in situ
Concept Barrett’s
Cancer
R. Kiesslich and M. I. Canto, GI Endoscopy Clinics N Am 19(2):261-272(2009).
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(Active) Neuron / Brain Function Imaging(ultimately on freely walking animals)
20um
Vessel
Astrocyte?
Astrocyte (star-shaped glial cells): provision of nutrients, maintenance of extracellular ion balance,long-distance propagation of Ca+2 wave, etc.
Endomicroscope
Freely-walking animal
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Basic Building Blocks
Core
Inner Cladding
Outer Cladding
Double-clad Fiber: Delivery and Collection
Photonic Crystal Fiber (Dispersion Management)
Miniature Scanning Head
DCF with MMFPZT(b)
5 mm
Compound lens
Resonant PZT Scanner (1mm, kHz)Miniature Compound Lens
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Rapid Spiral Scanning Pattern
X. M. Liu, M. J. Cobb, M. B. Kimmey, and X. D. Li, Optics Letters (2004)Y. C. Wu, Y. Leng, J. F. Xi, and X. D. Li, Optics Express (2009).L. Huo, J. F. Xi, Y. C. Wu, and X. D. Li, Optics Express (2010).
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Microendoscope
fs Fiber Laser
12”
18”
7”
Photo of TPF/SHG Endomicroscope
The whole system is very compact!
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TPF Images of GFP Mouse Tissues (ex = 870nm)
Pancreas20um 20um
Liver20um
Salivary Gland
20umKidney
20umBrain
20umSpleen
Collaboration with Dr. R. Weigert at NIH
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20µm
Intrinsic TPF Endomicroscopy of Fresh Tissues
Fresh Pig Esophagus
Fresh Rat Oral Cavity Fresh Rat LipFresh Mouse Dorsal Skin
Fresh Mouse Xenograft TumorFresh Mouse Colon
20µm
20µm20µm
20µm20µm60mW 60mW
60mW
60mW
50mW 50mW
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Further Comparison of Endomicroscopy with Microscopy for SHG Imaging of Breast Tissues
10 μm 10 μm10 μm 10 μm
10 μm10 μm 10 μm10 μm
Endo EndoMicroscopy Microscopy
No
rmal
Tu
mo
r
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Thank you!