The method proposed in this work enables accurate registration (on the scale of a few cells) of virtual slides of adjacent tissue sections on which the expression of different proteins is evidenced by standard IHC. Furthermore, combining our method with a sequential labeling and erasing technique enables cell-scale colocalization.
One of the most widely used techniques to obtain anatomical information is magnetic resonance imaging (MRI). Despite its high resolution, it has a low sensitivity which could be enhanced by coupling MRI with a more sensitive technique such as photoacoustic imaging (PAI). The development of a bimodal agent could thus lead to hybrid images with a high anatomical resolution provided by MRI and a precise localization of the contrast agent thanks to PAI. The probes used in this work [a] NMR and Molecular Imaging, 3354 Scheme 2. Synthetic pathway to obtain the pyridine compound 3. Reagents and conditions: (a) see ref (24); (b) K 2 CO 3 , EtOH, H 2 O, r.t., 3 h; (c) BrCH 2 CO 2 Et, DMF, 80°C, 1 h30; (d) CBr 4 , PPh 3 , CH 3 CN, 0°C to r.t., overnight.
3356Scheme 4. Synthesis of the precursor of the PCTA-based chelating agent. Reagents and conditions: (g) Na 2 CO 3 , CH 3 CN, r.t., 2 d; (h) NaOH, EtOH, H 2 O, r.t., 1 h30.
Superparamagnetic iron oxide nanoparticles (SPION) are very promising contrast media, especially for molecular imaging, due to their superior NMR efficacy. They even have wider biomedical applications such as in drug and gene delivery, tissue engineering and bioseparation, or as sensitive biological nanosensors. By coupling them to affinity ligands, SPION can bind to drugs, proteins, enzymes, antibodies or nucleotides. For in vitro biomedical applications, the detection of molecular interaction is possible by using a diversity of systems capable of sensing the magnetic properties of these materials. The goal of the present work was to develop and validate various in vitro biomedical applications of ultrasmall superparamagnetic particles of iron oxide (USPIO), including some that are not related to their magnetic properties. USPIO coated with dextran, starch or bisphosphonate exposing carboxylate groups were synthesized and some of them were functionalized by conjugating various biomolecules, such as biotin, streptavidin and apoptosis, or VCAM-1 specific peptides. The in vitro biomedical applications assessed in the present work included: (1) the relaxometric measurement of antibody concentration, cell receptor expression, molecular interaction, and enzymatic activity in aqueous suspensions; (2) MRI visualization of cells and detection of molecular interaction in an ELISA system; (3) ELISA applications of USPIO derivatives; and (4) detection of specific biomolecules by histochemistry. Our results confirm that rapid and simple in vitro detection of a diversity of functionalized SPION with relevance in medicine is possible by the existing NMR techniques and by chemical staining reactions. The protocols can be applied to minimally prepared biological samples (e.g. whole blood, blood plasma or serum, cell suspensions, biopsies, histological preparations, etc.), and often do not need complicated systems of signal amplification. The use of SPION labeled compounds could furthermore contribute to cost reductions in the diagnosis and in patient care.
Building further upon the high spatial resolution offered by ultrasonic imaging and the high optical contrast yielded by laser excitation of photoacoustic imaging, and exploiting the temperature dependence of photoacoustic signal amplitudes, this paper addresses the question whether the rich information given by multispectral optoacoustic tomography (MSOT) allows to obtain 3D temperature images. Numerical simulations and experimental results are reported on agarose phantoms containing gold nanoparticles and the effects of shadowing, reconstruction flaws, etc. on the accuracy are determined.
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