in-vivo real time non-invassive imaging for longitudinal ... · in-vivo real time non-invassive...

In-vivo real time non-invassive imaging for longitudinal translational research

Milan Kopecek

Regional Sales Manager

• Micro-ultrasound

• What is Micro-ultrasound?

• Visualization, measurement and analysis with the Vevo 2100

• Photoacoustics

• What is Photoacoustics?

• Visualization, measurement and analysis with the Vevo LAZR

• Applications

Agenda

Total Small Animal Solution

• Real-time

• Longitudinal Studies

• Measure Physiological parameters

• Contrast/Molecular Imaging

• Translatable to man

How Does Micro-Ultrasound Work?

How Does Micro-Ultrasound Work?

Micro-ultrasound (mouse fetus)

High-frequency = High-resolution

3 - 15 MHz

20cm

30 – 80 MHz

3cm

Melon

Coffee Bean

Conventional clinical ultrasound (human fetus)

Micro-Ultrasound: Clarity

200 – 300 micron resolution

30 micron resolution

Anatomy and Morphology

Cardiac Imaging: EKV

• High Temporal Resolution Left Ventricle in Adult Mouse at

10,000 frames per second

Visualization Measurement Analysis

• Tissue/organ/tumor identification

• Vasculature density, structure and flow

• Biomarkers

• Linear, area and volume measurements

• Blood flow velocity and vessel density

• Cardiac measurements

• Contrast Quantification including biomarker expression

• Heart wall motion and strain

Imaging with the Vevo

Visualization with Micro-Ultrasound

1 mm *WAP-TAg Mouse Model

Image courtesy of Cotarla I. and Furth, P., Lombardi Cancer Centre, Georgetown University

Early detection and Quantification of Pre-Palpable Mammary Tumors

Image courtesy of T. Minko, Rutgers, The State University of New Jersey

Video Loop

3D Quantification of Human Ovarian Tumor Model

Power Doppler Mode Imaging

Day 1 Day 4

PV = 3.18% PV = 11.22%

Images courtesy of Hastie, Chambers, Lacefield and Fenster; Robarts Research Institute, London

Precise interventions without the need for invasive surgery

Ultrasound Image-Guided Injections

Measurement with Micro-Ultrasound

Quantification of size, vasculature and perfusion

Early detection/screening

Perfusion Volumetrics

Vascularity

Cardiovascular measurements

LV analysis

Vascular stiffness

Blood flow

Strain

Analysis with Micro-Ultrasound

User defined regions of interest

Curve fit algorithms

Vevo MicroMarker Quantification

Velo

city D

isplacem

ent

Strain

S

train R

ate

Kovacs, Washington University

VevoStrainTM

• Micro-ultrasound

• What is Micro-ultrasound?

• Visualization, measurement and analysis with the Vevo 2100

• Photoacoustics

• What is Photoacoustics?

• Visualization, measurement and analysis with the Vevo LAZR

• Applications

Agenda

The Photoacoustic Effect

1) Nanosecond laser pulse illumination

3) Emitted pressure (sound) wave

4) Detection of ultrasound and creation of image

2) Optical absorption, heating and thermoelastic expansion

Optical Ultrasound

The Photoacoustic Effect

Light in

Sound out

5 mm 5 mm

5 mm

Meaningful images with the sensitivity of optical imaging and the resolution of ultrasound

The Photoacoustic Imaging

Visualization Measurement Analysis

• Blood signal

• Vascular density, structure

• Angiogenesis

• Oxygen saturation and total hemoglobin

• Photoacoustic signal intensity (contrast agent measurement)

• Absorption spectra

• Spectral unmixing

Imaging with the Vevo LAZR Photoacoustic System

Visualization with Photoacoustics

• Inherent co-registration of photoacoustic and anatomical images – Simultaneous registration of photoacoustic image on 2D and 3D planes – Real-time acquisitions for true in vivo monitoring

What is this signal?

Where is this signal?

Needle Skin Tumor Needle Skin Tumor

Delivery of nanoparticles into tumor

The Photoacoustic Imaging

Tumor vasculature imaging

Single wavelength imaging of endogenous signal

•Sensitive detection of blood signal

•Functional and non-functional vessels, blood pools

•2D and 3D

Measurement with Photoacoustics

( HbT = HbO2 + Hb )

Total Hemoglobin = Oxygenated Hemoglobin + Deoxygenated Hemoglobin

Near-Infrared (NIR)

Oxygen Saturation Calculation

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10

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30

40

50

60

70

80

90

0 100 200

Oxyg

en

Satu

rati

on

(%

)

Time (s)

Region 1 Region 2

100% O2 100% O2 5% O2

Change in Oxygen Saturation in Tumor

Region 1

Region 2

Analysis with Photoacoustics

Endogenous contrast

Contrast

Agent

Applications

Vevo LAZR Contrast Applications

Absorption Spectra Analysis – Spectro Imaging

Indocyanine green absorption spectra measured by photoacoustic imaging from 680 to 970 nm wavelengths.

Applications of Ultrasound and Photoacoustics

Single Walled Carbon Nanotubes conjugated to RGD peptide target αvβ3 integrins

(Gambhir lab, Stanford University)

(Heinmiller, et al. EMIM 2011)

Targeted carbon nanotubes

Change in Measured Photoacoustic Signal over time after carbon nanotube injection

Normal

Color Doppler Oxygen

saturation Total

hemoglobin

Stroke 2

Stroke 1

Stroke Imaging – Skin removed

Melanin

Oxygenated blood

3D Imaging of Human Finger

For more info contact us at info@visualsonics.com

milan.kopeck@fujifilm.com

Thank You

Pharmacokinetics with Photoacoustics

Identifying IR800-2DG in the kidney

Peak ~785nm Peak 760nm

Control and kidney

regions have peak PA

signal at different

wavelenths

Peak ~785nm

IR800-2DG in phantom

Spectral unmixing

Pre

24hrs post

6hrs post

Blue = DeoxyHb Red = OxyHb Green = IR800 2_DG

Quantifying IR800-2DG in the kidney

0

0.2

0.4

0.6

0.8

1

1.2

1.4

Pre 6 hrs post 24 hrs post

3D quantification of PA signal in the kidney pre, 6hrs and 24hrs post-IR800-

2DG

780nm 850nm

Methylene blue i.v. Injection – kidney imaging • Subtraction image of bolus injection of methylene blue using the ‘invisible

table’ and trifurcated fiber

Methylene blue i.v. Injection – kidney imaging

Pre-bolus

7 mins Post-bolus

Methylene blue i.v. Injection – kidney imaging

Reporter gene imaging with Photoacoustics

Photoacoustic Imaging of an Inducible Reporter

• Tyrosinase, the enzyme responsible for melanin production was used as an inducible reporter

• Animals with TYR- and TYR+ tumors on each flank were imaged before and after induction of tyrosinase expression

Abs.

Figure modified from Murkin and Arango. Near-infrared spectroscopy

as an index of brain and tissue Oxygenation. Br J Anaesth 2009; 103 (Suppl. 1): i3–i13

-TYR

+TYR

Absorption spectra for hemoglobin and

melanin

Post-induction Pre-induction

TYR-

a)

Photoacoustic Imaging of an Inducible Reporter

TYR+

a)

Photoacoustic Imaging of an Inducible Reporter

a)

Multi-modality Molecular Imaging with Tyrosinase as a Reporter Gene

MRI PET

PA

Figures from Cheng, Z. et al. Tyrosinase as a multifunctional reporter gene for Photoacoustic/MRI/PET triple modality molecular imaging. Sci Rep. 2013 Mar 19;3:1490

Imaging Enzymatic Activity

Pre

Post

680 nm 750 nm

Min

Max Pre Injection

680-750

Post Injection

680-750

Subtraction of Images

Levi, J. et al. Clin Cancer Res, 2013

MSCs + NTs Control MSCs + NTs Control

MSCs + NTs

in vivo monitoring MSC’s labeled with nanotracers

Nam et al SPIE 2012

Ischemic muscle

PEGylated fibrin gel

MSCs labeled with

NTs

US/PA imaging

PEGylated fibrin gel with MSCs +

NTs (105 cells/mL)

MSCs+NTs

Skin

HbO2

Hb

Longitudinal in vivo monitoring MSC’s labeled with nanotracers

Nam et al SPIE 2012

PEGylated fibrin gel with MSCs +

NTs

DAY 0 DAY 3 DAY 10 DAY 7

Injection

3D combined ultrasound and spectroscopic images of the gastrocnemius in which the

PEGylated fibrin gel containing MSCs/NTs was injected (Day 3,7, and 10)

23 mm x 12.5 mm x 25 mm

Neuroimaging with Photoacoustics

Brain imaging with photoacoustics

• Mouse brain coronal section

Non-Invasive 2D Brain Imaging with Oxyhemo Mode – Prototype LZ201

PCA

PCommA.

ICA

ICA

Noninvasive imaging of mouse brain

From: Dorr, et al. Neuroimage, 2007

Non-Invasive 3D Brain Imaging with Oxyhemo Mode

Superior sagittal sinus

Inferior cerebral

vein

Skin intact Skin intact

OxyZated Oxygen saturation map

HemoMeaZure Total hemoglobin map

1mm 1mm

Imaging of mouse brain ventricles

LZ250 LZ550

Methylene Blue

Injection site

Imaging of mouse brain ventricles

MRI microscope Multiplexed Vevo LAZR

PA image showing methylene blue in the ventricles

Van Hagen et al., Neurobiology of Disease, 2007.

Brain Tumor Imaging with Microultrasound and

Photoacoustics

Swanson Biotechnology Center Animal Imaging and Preclinical Testing Facility

Scott Floyd lab

Nonlienar contrast imaging of perfusion with microbubbles

Nonlinear contrast Mouse #1

Images courtesy of: Swanson Biotechnology Center Animal Imaging and Preclinical Testing Facility and laboratory of Scott Floyd, Koch Institute for Integrative Cancer Research, MIT

Nonlienar contrast imaging of perfusion with microbubbles

Nonlinear contrast

Hypoperfused area

Mouse #1

Contrast quantifiation (relative blood volume)

Images courtesy of: Swanson Biotechnology Center Animal Imaging and Preclinical Testing Facility and laboratory of Scott Floyd, Koch Institute for Integrative Cancer Research, MIT

Multi-modality imaging of hemodynamics in the brain

Potential hypoxic area Mouse #1

Perfusion Flow Oxygenation Images courtesy of: Swanson Biotechnology Center Animal Imaging and Preclinical Testing Facility and laboratory of Scott Floyd, Koch Institute for Integrative Cancer Research, MIT

Multi-modality imaging of hemodynamics in the brain

Mouse #1

Perfusion (Relative blood volume, flow estimates)

Flow (Percent vascularity)

Oxygenation (sO2, HbT)

Images courtesy of: Swanson Biotechnology Center Animal Imaging and Preclinical Testing Facility and laboratory of Scott Floyd, Koch Institute for Integrative Cancer Research, MIT

Mouse #1

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10 20 30 40 50 60

sO2

Avg

(%

)

Coronal slice # (caudal to rostral)

Avg sO2 in L and R Hemisphere (mouse 1) Caudal to rostral slices

L Hemisphere sO2

R Hemisphere sO2

Quantification of oxygen saturation in L vs R hemispheres

Images courtesy of: Swanson Biotechnology Center Animal Imaging and Preclinical Testing Facility and laboratory of Scott Floyd, Koch Institute for Integrative Cancer Research, MIT

Nonlinear contrast Mouse #2

Nonlienar contrast imaging of perfusion with microbubbles

Images courtesy of: Swanson Biotechnology Center Animal Imaging and Preclinical Testing Facility and laboratory of Scott Floyd, Koch Institute for Integrative Cancer Research, MIT

Nonlinear contrast

Hypoperfused area

Mouse #2

Hyperperfused area

Hypoperfused area

Contrast quantifiation (relative blood volume)

Nonlienar contrast imaging of perfusion with microbubbles

Images courtesy of: Swanson Biotechnology Center Animal Imaging and Preclinical Testing Facility and laboratory of Scott Floyd, Koch Institute for Integrative Cancer Research, MIT

Mouse #2 Potential hypoxic area

Perfusion Flow Oxygenation

Multi-modality imaging of hemodynamics in the brain

Images courtesy of: Swanson Biotechnology Center Animal Imaging and Preclinical Testing Facility and laboratory of Scott Floyd, Koch Institute for Integrative Cancer Research, MIT

Mouse #2

Perfusion (Relative blood volume, flow estimates)

Flow (Percent vascularity)

Oxygenation (sO2, HbT)

Multi-modality imaging of hemodynamics in the brain

Images courtesy of: Swanson Biotechnology Center Animal Imaging and Preclinical Testing Facility and laboratory of Scott Floyd, Koch Institute for Integrative Cancer Research, MIT

Mouse #2

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50

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10 20 30 40 50

sO2

Avg

(%

)

Coronal slice # (caudal to rostral)

Avg sO2 in L and R Hemisphere (mouse 2) Caudal to rostral slices

L Hemisphere sO2

R Hemisphere sO2

Quantification of oxygen saturation in L vs R hemispheres

Images courtesy of: Swanson Biotechnology Center Animal Imaging and Preclinical Testing Facility and laboratory of Scott Floyd, Koch Institute for Integrative Cancer Research, MIT

Stroke imaging with Microultrasound and

Photoacoustics

Stroke vs normal animal - quantification

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10

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90

Stroke 1 Stroke 2 Normal

sO

2 (

%)

3D oxygen saturation

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5000

10000

15000

20000

25000

Stroke 1 Stroke 2 Normal

Hb

T (

a.u

.)

3D total hemoglobin

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2%

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10%

12%

14%

16%

Stroke 1 Stroke 2 Normal

PV

(%

)

3D percent vascularity (color Doppler)

Human imaging with Microultrasound and

Photoacoustics

Human Imaging

• Ultrasound/Photoacoustic transducer mounted to a 3D stepper motor

3D Imaging of Human Vasculature

• 3D MIP of the palm • 3D MIP of the forearm

• 3D MIP of the index finger

40mm

20mm

40mm

20mm

• 2D image of the forearm

Finger Imaging - Ischemia

• Dental floss tied around index finger to induce ischemia

• 2D imaging of finger ischemia – Oxyhemo mode

Finger Imaging - Ischemia

• 2D imaging of finger ischemia –Spectro mode

Abs.

Figure modified from Murkin and Arango. Near-infrared spectroscopy as an index of brain and tissue Oxygenation. Br J Anaesth 2009; 103 (Suppl. 1): i3–i13

Imaging of ???

Doppler imaging of dermatofibroma

Photoacoustic imaging of dermatofibroma

Photoacoustic Imaging Co-registered High-resolution High-sensitivity Real-time Oxygen saturation Molecular imaging In Vivo In Situ

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VisualSonics, VisualSonics logo, VisualSonics dot design, Vevo, Vevo MicroMarker, VevoStrain, VevoCQ, SoniGene, RMV, EKV, MicroScan, LAZRTight, Insight through In Vivo Imaging, are registered trademarks (in some jurisdictions) or unregistered trademarks of VisualSonics Inc. © 2011 VisualSonics Inc. All rights reserved.