Silicon detectors for combined MR-PET and MR-SPECT imaging.

Silicon based devices can extend PET-MR and SPECT-MR imaging to applications, where their advantages in performance outweigh benefits of high statistical counts.Silicon is in many ways an excellent detector material with numerous advantages, among others: excellent energy and spatial resolution, mature processing technology, large signal to noise ratio, relatively low price, availability, versatility and malleability. The signal in silicon is also immune to effects of magnetic field at the level normally used in MR devices. Tests in fields up to 7 T were performed in a study to determine effects of magnetic field on positron range in a silicon PET device. The curvature of positron tracks in direction perpendicular to the field's orientation shortens the distance between emission and annihilation point of the positron. The effect can be fully appreciated for a rotation of the sample for a fixed field direction, compressing range in all dimensions. A popular Ga-68 source was used showing a factor of 2 improvement in image noise compared to zero field operation. There was also a little increase in noise as the reconstructed resolution varied between 2.5 and 1.5 mm.A speculative applications can be recognized in both emission modalities, SPECT and PET.Compton camera is a subspecies of SPECT, where a silicon based scatter as a MR compatible part could inserted into the MR bore and the secondary detector could operate in less constrained environment away from the magnet. Introducing a Compton camera also relaxes requirements of the radiotracers used, extending the range of conceivable photon energies beyond 140.5 keV of the Tc-99m.In PET, one could exploit the compressed sub-millimeter range of positrons in the magnetic field. To exploit the advantage, detectors with spatial resolution commensurate to the effect must be used with silicon being an excellent candidate. Measurements performed outside of the MR achieving spatial resolution below 1 mm are reported.

Silicon as an Unconventional Detector in Positron Emission Tomography.

Positron emission tomography (PET) is a widely used technique in medical imaging and in studying small animal models of human disease. In the conventional approach, the 511 keV annihilation photons emitted from a patient or small animal are detected by a ring of scintillators such as LYSO read out by arrays of photodetectors. Although this has been a successful in achieving ~5mm FWHM spatial resolution in human studies and ~1mm resolution in dedicated small animal instruments, there is interest in significantly improving these figures. Silicon, although its stopping power is modest for 511 keV photons, offers a number of potential advantages over more conventional approaches. Foremost is its high spatial resolution in 3D: our past studies show that there is little diffculty in localizing 511 keV photon interactions to ~0.3mm. Since spatial resolution and reconstructed image noise trade off in a highly non-linear manner that depends on the PET instrument response, if high spatial resolution is the goal, silicon may outperform standard PET detectors even though it has lower sensitivity to 511 keV photons. To evaluate silicon in a variety of PET "magnifying glass" configurations, an instrument has been constructed that consists of an outer partial-ring of PET scintillation detectors into which various arrangements of silicon detectors can be inserted to emulate dual-ring or imaging probe geometries. Recent results have demonstrated 0.7 mm FWHM resolution using pad detectors having 16×32 arrays of 1.4mm square pads and setups have shown promising results in both small animal and PET imaging probe configurations. Although many challenges remain, silicon has potential to become the PET detector of choice when spatial resolution is the primary consideration.

Timing performance of the silicon PET insert probe.

Simulation indicates that PET image could be improved by upgrading a conventional ring with a probe placed close to the imaged object. In this paper, timing issues related to a PET probe using high-resistivity silicon as a detector material are addressed. The final probe will consist of several (four to eight) 1-mm thick layers of silicon detectors, segmented into 1 x 1 mm(2) pads, each pad equivalent to an independent p + nn+ diode. A proper matching of events in silicon with events of the external ring can be achieved with a good timing resolution. To estimate the timing performance, measurements were performed on a simplified model probe, consisting of a single 1-mm thick detector with 256 square pads (1.4 mm side), coupled with two VATAGP7s, application-specific integrated circuits. The detector material and electronics are the same that will be used for the final probe. The model was exposed to 511 keV annihilation photons from an (22)Na source, and a scintillator (LYSO)-PMT assembly was used as a timing reference. Results were compared with the simulation, consisting of four parts: (i) GEANT4 implemented realistic tracking of electrons excited by annihilation photon interactions in silicon, (ii) calculation of propagation of secondary ionisation (electron-hole pairs) in the sensor, (iii) estimation of the shape of the current pulse induced on surface electrodes and (iv) simulation of the first electronics stage. A very good agreement between the simulation and the measurements were found. Both indicate reliable performance of the final probe at timing windows down to 20 ns.

Monoclonal antibody against DNA adducts with osmium structural probes.

Osmium tetroxide complexes with nitrogen ligands (Os,L) have been widely used as probes of the DNA structure. A monoclonal antibody OsBP7H8 against DNA adducts with Os,L was produced in mice. OsBP7H8 does not bind to proteins or total yeast RNA modified with Os,2,2'-bipyridine (bipy) nor to the unmodified nucleic acids and proteins. The antibody recognizes DNA modified with Os,bipy (DNA-Os,bipy) or with OsO4,1,10-phenanthroline (DNA-Os,phen) but it does not cross-react with oxidized DNA and with DNA adducts of osmium tetroxide complexes with other ligands (such as pyridine, TEMED and bathophenanthroline disulfonic acid). The affinity of OsBP7H8 to DNA-Os,phen is about five-fold higher as compared to DNA-Os,bipy. The antibody can be thus applied either for recognition of single-stranded and distorted regions in DNA (after DNA modification with Os,bipy) or for detection of both single-stranded and double-stranded DNAs (after DNA modification with Os,phen). A new simplified procedure for the dot-blot analysis is proposed, not requiring the purification of DNA-osmium adduct prior to its application to the membrane.

Two superhelix density-dependent DNA transitions detected by changes in DNA adsorption/desorption behavior.

The adsorption behavior of covalently closed circular plasmid DNA at the mercury/water interface was studied by means of AC impedance measurements. The dependence of the differential capacitance (C) of the electrode double layer on the potential (E) was measured in the presence of adsorbed DNA. It was found that the C-E curves of supercoiled DNA at native and highly negative superhelix densities (sigma), relaxed covalently closed circular DNA, and nicked DNA differed from each other. A detailed study of topoisomer distributions ranging from -sigma of 0 to 0.11 revealed two supercoiling-dependent transitions, at about -sigma = 0.04 (transition TI) and 0.07 (transition TII). Transition TI was detected by measuring the height of the adsorption/desorption peak 1 (at about -1.2 V against the saturated calomel electrode) and the decrease of capacitance (DeltaC) at -0.35 V. This transition may be due to a sudden change in the ability of the DNA to respond to the alternating voltage, probably caused by changes in the DNA tertiary and/or secondary structure. Transition TII was detected by measuring peak 3* (at about -1.3 V), which was absent in topoisomers with -sigma less than 0.05. This transition is due to changes in the DNA adsorption/desorption behavior related to increased accessibility of bases at elevated negative superhelix density. Opening of the duplex at highly negative superhelix density was also detected by the single-strand selective probe of DNA structure, osmium tetroxide, 2, 2'-bipyridine. Our results suggest that electrochemical techniques provide sensitive experimental analysis of changes in DNA structure.

A supercoiled DNA-modified mercury electrode-based biosensor for the detection of DNA strand cleaving agents.

DNA-damaging agents in the environment represent a serious danger to human health. We use a supercoiled DNA-modified mercury electrode as a fast-response biosensor for the detection of DNA strand cleaving agents. The sensor is based on a strong difference between the a.c. voltammetric responses of covalently closed circular (supercoiled) and of open circular (nicked) plasmid DNA. We show that the sensor can detect hydroxyl radicals in laboratory-prepared solutions and in various natural and industrial water samples. The sensor is also capable of detecting unknown DNA-damaging agents in industrial waters.

Tumor suppressor protein p53 binds preferentially to supercoiled DNA.

Wild type human tumor suppressor protein p53 (expressed in insect cells) binds strongly to negatively supercoiled (sc) plasmid DNA at a native superhelix density, as evidenced by electrophoretic retardation of scDNA in agarose gels and imaging by scanning force microscopy (SFM). The binding occurs both in the presence and absence of the p53 consensus sequence. At relatively low p53/DNA ratios, binding of p53 to scDNA results in the appearance of several retarded DNA bands on the gels, similar to a conventional topoisomer ladder generated enzymatically. However, after removal of p53 by deproteination, the original mobility of the scDNA is recovered, indicating that the reduction of torsional stress accompanying p53 binding does not reflect changes in linking number. In DNA samples partially relaxed by topoisomerase I p53 binds preferentially to the scDNA molecules with the largest negative superhelix density. SFM imaging of the p53/scDNA complex reveals a partial or total relaxation of the compact scDNA, the degree of which increases with the number of bound p53 molecules. Competition assays with linear DNA reveal a preference of p53 for scDNA. In addition, scDNA induces dissociation of p53 from a preformed complex with a DNA fragment (474 bp) containing the consensus sequence. We conclude that the affinity of p53 for negatively supercoiled DNA is greater than that for the consensus sequence in linear fragments. However, thermally denatured linearized plasmid DNA is efficient in competing for the binding of p53 to scDNA, although the first retarded band (presumed to contain one bound p53 molecule) is retained in the case of the plasmid containing the consensus sequence. Thus, it appears that interactions involving both the core domain and the C-terminal domain regulate the binding of p53 to scDNA. The above results are not restricted to human p53; the wild type rat p53 protein also results in the retardation of scDNA on agarose gels. The biological implications of the novel DNA binding activities of p53 are discussed.