Molecular Imaging Probes

Molecular Imaging Probes

Molecular Imaging

A goal of the Meade lab is the development of magnetic resonance imaging (MRI) probes capable of reporting on anatomical and physiological function of cellular processes in whole animals.  MRI has proven to be an effective modality for molecular imaging applications by providing excellent spatial and temporal resolution and unlimited depth of penetration without the use of ionizing radiation. The bioactivated Gd(III) complexes prepared in this research are designed to report on a biological event of interest and transduce this metabolic signal and/or cellular status as a change in MR contrast. Additionally, targeted Gd(III) probes are being prepared to report cellular status through signal enhancement as a function of receptor expression. Our lab addresses the inherent sensitivity issues associated with MRI by developing probes that increase cellular uptake, thereby maximizing Gd(III) loading using both molecular and nano-based approaches. Development of new MR probes addresses an unmet need for monitoring specific tissue types and environments relevant to dynamic biological processes.

Our approach begins with computation and synthesis and proceeds to biological assays, in vivo imaging and processing.


q-modulated MR Probes

Bioactivated q-modulated contrast agents alter the coordination environment of the Gd(III) center as a means to modulate contrast. These probes allow for real-time in vivo imaging of dynamic cellular processes such as gene expression and ion fluxes.
In response to the biological event of interest, q-modulated contrast agents transform from a low-relaxivity q=0 state to a high relaxivity q=1 state.Lanthanide probes for bioresponsive imaging. Chem Rev 2014, 114, 4496.


alpha and beta egadme
Two configurations of a beta-galactosidase activated CA. Sugar configuration is dependent on the position of the methyl group either a or b to the macrocyclic core. Mechanistic investigation of beta-galactosidase-activated MR contrast agents. Inorganic chemistry 2008, 47, 56. 

The Meade lab pioneered the development of q-modulated Gd(III) contrast agents with Egad. In the absence of the reporter protein β-galactosidas (β-gal), the pendant sugar blocks water access to the Gd(III) center. Hydrolysis of the glycosidic bond by β-gal results in a change in the coordination environment (q) resulting in enhanced image contrast. Several generations of enzyme-activated agents have been developed with increased changes in contrast and improved kinetics of activation.

Embryos were injected at the two-cell stage. Both cells received EgadMe; one cell which represents the right (R) side of the animal also received mRNA for β-gal. (A) GFP florescence image of a living embryo. (B) MR image of the same living embryo in (A) with signal from water made transparent. (C) same embryo stained for
β-gal after MR imaging. These images demonstrate the noninvasive detection of regions positive for β-gal within a single living X. laevis embryo by MR.  A ”smart” magnetic resonance imaging agent that reports on specific enzymatic activity. Angewandte Chemie-International Edition in English 1997, 36, 726.

The current generation of β-gal-activated agents incorporate a self-immolative prodrug electron cascade and a back-binding carboxylate. This design has two major advantages: 1) the self-immolative electron cascade allows for rapid kinetics of activation and 2) the back-binding moiety makes the agent q=0 thus the “off” state is darker.

Current generation of beta-galactosidase-activated CAs. The self-immolative electron cascade allows for fast kinetics while the back-binding carboxylate (in red) efficiently blocks water prior to activation. A gadolinium chelate for detection of beta-glucuronidase: a self-immolative approach. Journal of the American Chemical Society 2005, 127, 12847.

Ion-activated agents:  Zn(II) and Ca(II)

Ca(II) agent
Calcium binding the BAPTA domain of the Gd(III)-DOPTA agent. This mechanism mechanism of activation is sensitive to millimolar changes in Ca(II) concentration. A calcium-sensitive magnetic resonance imaging contrast agent. Journal of the American Chemical Society 1999, 121, 1413.  Mechanistic studies of a calcium-dependent MRI contrast agent. Inorganic chemistry 2002, 41, 4018.

We have developed q-modulated Gd(III) contrast agents responding selectively and specifically to Zn(II) and Ca(II).  Calcium is the most important intracellular secondary messenger in the body. Spatial and temporal detection of Ca(II) fluxes provides insight into many biological mechanisms. The Meade lab developed the first ion sensing q-modulated contrast agent. The Ca(II) agent is comprised of a aminopolycarboxylic acid Ca(II)-binding ligand known as BAPTA.  In the absence of Ca(II), the acetate groups of BAPTA are believed to bind to the Gd(III) center, restricting water access. Ca(II) concentrations in the millimolar range are bound by the BAPTA domain resulting in an increase in MR signal.

The optimal linker length(s) n=4/5 was determined by VT-NMR studies of 13C labeled carboxylates of the Eu analogue. Mechanisms of ZnII-activated magnetic resonance imaging agents. Inorganic chemistry 2008, 47, 10788. Structural optimization of Zn(II)-activated magnetic resonance imaging probes. Inorganic chemistry 2013, 52, 12250.

Zn(II) is the second most abundant transition metal in the body with known structural and signaling roles, similar to Ca(II). A structurally optimized Zn(II) responsive agent has been developed to respond to changes in millimolar concentrations of Zn(II).

Targeted MR-Probes 


HaloTag – targeted CA bound to extracellular HaloTag.

The receptor enhanced magnetization enhancement (RIME) effect is a common strategy to increase contrast in an MR image. RIME relies on the coupling of CA motion to a macromolecule or protein, thereby increasing its rotational correlation time; lengthening of this value at low magnetic fields translates to an increase in image brightness. To this end, a haloalkane dehalogenase (HaloTag gene reporter protein) targeted MR probe was developed. HaloTag selectively and irreversibly binds this agent’s distal haloalkane, enabling MR molecular imaging of gene expression with high specificity and sensitivity.


Determination of progesterone (PR) status in hormone-dependent diseases such as ovarian and breast cancer is essential for assessing disease states and subsequent decisions regarding treatment. The Meade lab is developing noninvasive PR-targeted MR probes to monitor PR status, precluding the need for biopsy. The first generation (ProGlo) of this agent showed selective accumulation in PR+ tumors, but was limited due to its low solubility. We are currently working towards water-soluble agents that maintain the same selective accumulation.

Structure of PR targeted MR contrast agent.
Structure of PR targeted MR contrast agent (ProGlo). The construct consists of a 21-hydroxy progesterone targeting group, a 6C linker, and a Gd(III) DO3A chelate.  Synthesis and biological evaluation of water-soluble progesterone-conjugated probes for magnetic resonance imaging of hormone related cancers. Bioconjugate chemistry 2011, 22, 2304. Rational design, synthesis, and biological evaluation of progesterone-modified MRI contrast agents. Chem Biol 2007, 14, 824. A steroid-conjugated contrast agent for magnetic resonance imaging of cell signaling. Journal of the American Chemical Society 2005, 127, 13164.
Bilateral flank tumors were grown in a mouse model PR+ and PR-. ProGlo selectively accumulates in the PR+ tumor allowing versus the blood pool agent Gd-DO3A.
Bilateral flank tumors (PR+ and PR-) were grown in a mouse model. ProGlo selectively accumulates in the PR+ tumor allowing versus the blood pool agent Gd-DO3A.  A steroid-conjugated magnetic resonance probe enhances contrast in progesterone receptor expressing organs and tumors in vivo. Mol Pharm 2011, 8, 1390. Progesterone-Targeted Magnetic Resonance Imaging Probes. Bioconjugate chemistry 2014.

Lipophilic CAs

Lipophilic CAs incorporate into the cell membrane for tracking cells in vivo by MRI.
Lipophilic CAs incorporate into the cell membrane for tracking cells in vivo by MRI. Cell labeling via membrane-anchored lipophilic MR contrast agents. Bioconjugate chemistry 2014, 25, 945.

A series of lipophilic MR CAs have been developed for cell-tracking in vivo by labeling the cells ex vivo.  Rationale design of these agents incorporates the fatty acids commonly incorporated in cellular membranes conjugated to a Gd-DO3A chelate.  These new agents exhibit significantly enhanced labeling and retention in HeLa and MDA-MB-231-mcherry cells compared to agents that are internalized by cells.

Multimeric MR-optical CAs 

The agent consists of three macrocyclic Gd(III) chelates conjugated to a fluorophore and possesses high relaxivity, water solubility, and is nontoxic. The modular synthesis is amenable for the incorporation of a variety of fluorophores to generate molecular constructs for a number of applications.

Bimodal fluorescent-MR active CA. A multimeric MR-optical contrast agent for multimodal imaging. Chem Commun (Camb) 2014, 50, 11469.

Signal Amplification using Nanotechnology 

Nanotechnology provides a diverse set of multifunctional platforms for the development of MR contrast agents. The Meade lab uses two biologically-compatible nanoconstructs based on either carbon or gold. These constructs allow for high Gd(III) loading in addition to targeting moieties.

DNA-Au Nanoparticles

13 nm Gd(III)-enriched DNA-Au nanoparticle (AuNP) conjugates represent a new class of MR CAs that demonstrate efficient cell penetration and accumulation allowing for imaging small cell populations with μM Gd(III). These AuNPs also bear a pendant fluorophore that allows for histological analysis.

Gd(III)-Au-DNA nanoparticle synthesis. Citrate stabilized AuNPs conjugated with thiol-labeled 24-mer poly dT oligonucleotides. These single strands of DNA are modified with azides primed to perform a click reaction with an alkyne pendant Gd(III) chelate. Multimodal gadolinium-enriched DNA-gold nanoparticle conjugates for cellular imaging. Angewandte Chemie 2009, 48, 9143.

Gold Nanostars

In collaboration with the Odom group at Northwestern we are developing Gd(III)-DNA Au Nanostars. Using the same synthetic scheme as the spherical AuNPs, these faceted particles  enable large second-sphere water contributions to further enhance contrast efficiency.

DNA-Au Nanostars
Gd(III)-DNA-Au Nanostar synthesis, alkyne modified Gd(III) chelates are clicked onto a thiol-terminated 24mer polyT DNA strand modified with a fluorophore. The thiol end is conjugated to a Au-Nanostar.


Gd(III) associated with graphene relaxes water proton spins at an effectiveness that approaches or exceeds the theoretical limit for a single bound water molecule. These Gd(III)-labeled materials represent a potential breakthrough in sensitivity for Gd(III)-based contrast agents used in MRI.  A library of Gd(III) chelates have been evaluated for their relaxivity and cellular uptake properties.

Pictorial representation of Gd(III) MR chelate associated to graphene oxide nanoparticles. Mechanisms of Gadographene-Mediated Proton Spin Relaxation. The journal of physical chemistry. C, Nanomaterials and interfaces 2013, 117.


Gd(III)-nanodiamond conjugates have been prepared and exhibit a 10-fold increase in relaxivity relative  to the corresponding molecular species. Nanodiamonds provide an easily functionalized material with tunable therapeutic loading and release profiles.

Uptake of Gd(III)-nanodiamonds via endocytosis, demonstrating the advantages of this biocompatible nano material. Gd(III)-nanodiamond conjugates for MRI contrast enhancement. Nano letters 2010, 10, 484.

Visualization and Volume Rendering of Images

The images generated using these probes are displayed and interrogated in the Center for Advanced Molecular Imaging adjacent to our laboratory ( The 3D “Wall” is based on graphics Processing Units (GPUs) that were originally designed for the sole purpose of rendering computer generated imagery onto a digital display. Over the last decade, however, these increasingly powerful processors have evolved to solve problems they were not originally intended to solve. A growing number of commercial software products, including the digital artist tools used by Northwestern Visualization to generate animated movies, now integrate these GPUs into their workflow. Faculty, graduate students, and postdoctoral fellows have at their disposal an immersive, highly-interactive means of visually interrogating their data while simultaneously serving as a medium on which to present their three-dimensional data to colleagues, students, and the general public.



Chemistry at Northwestern University