Vivo Monitoring Research Articles

In Vivo Monitoring of Quantum Dots in the Extracellular Space Using Push−Pull Perfusion Sampling, Online In-Tube Solid Phase Extraction, and Inductively Coupled Plasma Mass Spectrometry

To monitor the dynamic changes of extracellular quantum dots (QDs) in vivo in the livers of anesthetized rats, we developed an automatic online analytical system comprising push-pull perfusion (PPP) sampling, the established in-tube solid phase extraction (SPE) procedure, and inductively coupled plasma mass spectrometry (ICPMS). The method takes advantage of the retention of QDs onto the interior surface of a polytetrafluoroethylene (PTFE) tube as a means of extracting the QDs from complicated push-pull perfusates. For the injected QDs present in the liver extracellular fluid (ECF) at low picomolar levels, a temporal resolution of 10 min was required to collect sufficient amounts of QDs to meet the sensitivity requirements of the ICPMS system. To the best of our knowledge, this study is the first to exploit the PPP technique for the collection of QDs from living animals and PTFE tubing as a SPE adsorbent for the online extraction of QDs and the removal of biological matrix prior to ICPMS analysis of cadmium-containing inorganic nanocrystal. We confirmed the analytical reliability of this method from measurements of the spike recoveries of saline samples; in addition, we demonstrated the systems' applicability through in vivo monitoring of the time-dependent concentration profile of liver extracellular QDs in living rats after intravenous administration.

Efficient and Rapid Labeling of Transplanted Cell Populations with Superparamagnetic Iron Oxide Nanoparticles Using Cell Surface Chemical Biotinylation for in Vivo Monitoring by MRI

Determination of the dynamics of specific cell populations in vivo is essential for the development of cell-based therapies. For cell tracking by magnetic resonance imaging (MRI), cells need to internalize, or be surface labeled with a MRI contrast agent, such as superparamagnetic iron oxide nanoparticles (SPIOs): SPIOs give rise to signal loss by gradient-echo and T(2)-weighted MRI techniques. In this study, cancer cells were chemically tagged with biotin and then magnetically labeled with anti-biotin SPIOs. No significant detrimental effects on cell viability or death were observed following cell biotinylation. SPIO-labeled cells exhibited signal loss compared to non-SPIO-labeled cells by MRI in vitro. Consistent with the in vitro MRI data, signal attenuation was observed in vivo from SPIO-labeled cells injected into the muscle of the hind legs, or implanted subcutaneously into the flanks of mice, correlating with iron detection by histochemical and X-ray fluorescence (XRF) methods. To further validate this approach, human mesenchymal stem cells (hMSCs) were also employed. Chemical biotinylation and SPIO labeling of hMSCs were confirmed by fluorescence microscopy and flow cytometry. The procedure did not affect proliferation and multipotentiality, or lead to increased cell death. The SPIO-labeled hMSCs were shown to exhibit MRI signal reduction in vitro and was detectable in an in vivo model. In this study, we demonstrate a rapid, robust, and generic methodology that may be a useful and practical adjuvant to existing methods of cell labeling for in vivo monitoring by MRI. Further, we have shown the first application of XRF to provide iron maps to validate MRI data in SPIO-labeled cell tracking studies.

In Vivo Imaging and Monitoring of Transplanted Stem Cells: Clinical Applications

Regenerative medicine using stem cells has appeared as a potential therapeutic alternative for coronary artery disease, and stem cell clinical studies are currently on their way. However, initial results of these studies have provided mixed information, in part because of the inability to correlate organ functional information with the presence/absence of transplanted stem cells. Recent advances in molecular biology and imaging have allowed the successful noninvasive monitoring of transplanted stem cells in the living subject. In this article, different imaging strategies (direct labeling, indirect labeling with reporter genes) to study the viability and biology of stem cells are discussed. In addition, the limitations of each approach and imaging modality (eg, single photon emission computed tomography, positron emission tomography, and MRI) and their requirements for clinical use are addressed. Use of these strategies will be critical as the different regenerative therapies are being tested for clinical use.

THE CANADIAN NATIONAL CALIBRATION REFERENCE CENTER FOR BIOASSAY AND IN VIVO MONITORING: AN UPDATE

The Canadian National Calibration Reference Center (NCRC) for Bioassay and In Vivo Monitoring is part of the Radiation Protection Bureau, Health Canada. The NCRC operates three performance testing programs that are designed to confirm that workplace monitoring results are accurate and provide the necessary external verification that is part of a comprehensive quality assurance program. The NCRC performance testing programs cover the in vitro, in vivo, and internal dosimetry parts of Canadian facilities' radiation protection programs. The internal dosimetry performance testing is a new addition to the performance testing suite. This summary also describes the recent reorganization of the NCRC.

In Vivo Monitoring of Venous Thrombosis in Mice.

Abstract 5061Venous thromboembolism afflicts 117 people per 100,000 each year and is an important cause of morbidity and mortality. There has been extensive research dedicated to the clinical aspect of venous thrombosis, especially with regards to its diagnosis and treatment. However, animal models studying this phenomenon are scarce and, in most cases, very crude. Developing a murine model of venous thrombosis using techniques similar to the ones used to detect thrombosis in humans can be a constructive step in studying this phenomenon in more detail. The model developed in our lab uses ultrasound imaging to visualize venous clots in the Inferior Vena Cava (IVC) of mice, allowing for precise measurements of the formed clot. Ligation of the IVC is one of the well established models for studying thrombosis in mice. We ligated the IVC of wild type C57B6 mice, and allowed them to recover. We then followed clot formation at several time points after the operation using micro-ultrasonography, the Vevo 770®, a novel imaging ultrasound technology designed to monitor murine vasculature. To assess the precision of the clot measurements, we then sacrificed the mice, and dissected out the thrombi in order to precisely measure and weigh them. A thrombosis develops only after 5 hours of ligation post surgery when a clot is visualized in the IVC. The clot increases slightly over the next 24 hours. The measurements of the clot after dissection correlates favourably with the measurements done by ultrasonagraphy using the Vevo770®. These data suggest that the Vevo770® can be used as a reliable technique for non-invasive assessment of venous thromboembolism in mice. Developing a murine model for thrombosis using more accurate, and clinically more relevant techniques such as ultrasonography, is a step towards better understanding and treatment of venous thromboembolism. DisclosuresNo relevant conflicts of interest to declare.

Direct Profiling of Phytochemicals in Tulip Tissues and In Vivo Monitoring of the Change of Carbohydrate Content in Tulip Bulbs by Probe Electrospray Ionization Mass Spectrometry

Probe electrospray ionization (PESI) is a recently developed ESI-based ionization technique which generates electrospray from the tip of a solid needle. In this study, we have applied PESI interfaced with a time of flight mass spectrometer (TOF-MS) for direct profiling of phytochemicals in a section of a tulip bulb in different regions, including basal plate, outer and inner rims of scale, flower bud and foliage leaves. Different parts of tulip petals and leaves have also been investigated. Carbohydrates, amino acids and other phytochemicals were detected. A series of in vivo PESI-MS experiments were carried out on the second outermost scales of four living tulip bulbs to monitoring the change of carbohydrate content during the first week of initial growth. The breakdown of carbohydrates was observed which was in accordance with previous reports achieved by other techniques. This study has indicated that PESI-MS can be used for rapid and direct analysis of phytochemicals in living biological systems with advantages of low sample consumption and little sample preparation. Therefore, PESI-MS can be a new choice for direct analysis/profiling of bioactive compounds or monitoring metabolic changes in living biological systems.

Application of Magnetoelastic Thick Films for Wireless, In Vivo Monitoring of Pressure at Abdominal Aortic Aneurysm

According to the National Center for Health Statistics, cardiovascular diseases remain the number one killer in the United States. Among the various types of cardiovascular diseases, the aortic aneurysm is ranked number nine. The abdominal aortic aneurysm (AAA), in particular, is an abnormal, localized dilation of the abdominal aorta wall caused by weakened or diseased aorta walls. One of the treatments for this disease is using an endovascular surgery at which an endovascular graft is delivered to the aneurysm site through the femoral arteries. The deployment of the endovascular graft will exclude blood flow to the aneurysm, thus preventing further expansion of the aneurysm sac. Although this technique is preferred over open surgeries due to its minimal invasiveness, an event known as the endoleak, where the endovascular graft fails to retain the blood and leads to leakage to the aneurysm sac, may occur. Here, we are developing a novel pressure monitoring system to remotely and continuously measure the pressure in the aneurysm sac. The main component of the system is a pressure-sensitive material, made of a magnetoelastic, magnetically soft film attached or coated on the endovascular graft. When under an AC magnetic field (excitation field), the magnetoelastic film generates a secondary magnetic field. Due to its magnetoelastic property, the amplitude of the secondary field varies with applied stresses, allowing remote pressure monitoring. To eliminate noises from the excitation field, the generated secondary field is measured at twice the excitation frequency to obtain the 2nd-order harmonic field, which is used for tracking the pressure variations. A scaled-up prototype of the pressure monitoring system was constructed and examined to demonstrate the feasibility of this technology. A commercial magnetoelastic thick film, Metglas 2826MB from Metglas, Inc., was attached on a polycarbonate substrate and covered by a thin polycarbonate protective layer. The substrate was then embedded in a plastic tube with flowing liquid to represent the condition of an aorta. Liquid pressure in the tube was altered during the experiment by restricting or relaxing the flow channel. In this study, a 10mm×40mm file (Film A) and a 5mm×40mm film (Film B) were fabricated and tested. The amplitude of the 2nd-order harmonic field produced by the films was inversely proportional to the fluid pressure. It was also shown that films with different sizes exhibited different signal sensitivity with the smaller film (Film B) exhibited greater sensitivity. This experiment indicates that feasibility of the pressure monitoring technology.

Noninvasive In Vivo Monitoring of Cyanide Toxicity and Treatment Using Diffuse Optical Spectroscopy in a Rabbit Model

Currently, no reliable noninvasive methods exist for monitoring the severity of in vivo cyanide (CN) toxicity, treatment, and resulting physiological changes. We developed a broadband diffuse optical spectroscopy (DOS) system to measure bulk tissue absorption and scattering. DOS was used to optically monitor CN toxicity and treatment with sodium nitrite (NaNO2). To perform experiments, the DOS probe was placed on the hind leg of rabbits. A sodium CN solution was infused intravenously. DOS and concurrent physiologic measurements were obtained. After completion of CN infusion, NaNO2 was infused to induce methemoglobinemia (MetHb). During infusion of CN, blood gas measurements showed an increase in venous partial pressure of oxygen (pO2), and following reversal, venous pO2 values decreased. DOS measurements demonstrated corresponding changes in hemoglobin oxygenation states and redox states of cytochrome-c oxidase (CcO) during CN infusion and NaNO2 treatment. Therefore, DOS enables detection and monitoring of CN toxicity and treatment with NaNO2.

Canadian national internal dosimetry performance testing programme: results of the pilot programme

This paper describes the design and construction of a new performance testing programme that was implemented in Canada in 2008. The Canadian Regulator (Canadian Nuclear Safety Commission) had determined that their licensees, in addition to participating in the existing in vivo and in vitro performance tests, needed to demonstrate their ability in interpreting bioassay results to obtain intakes and resulting doses. The new programme is administered by the Canadian National Calibration Reference Center for Bioassay and In Vivo Monitoring (NCRC). Currently, the NCRC carries out the performance testing for the in vitro and in vivo. At the time of writing, the first round has been completed and the results for (3)H and (nat)U exposures were very consistent, while the committed effective dose from (137)Cs intake varied by a factor of two.

In Vivo Monitoring of Angiogenesis Inhibitory Treatment Effects by Dynamic Contrast-Enhanced Computed Tomography in a Xenograft Tumor Model

To evaluate the potential of dynamic CT enhanced by iohexol or a novel macromolecular contrast agent, PEG12000-Gen4-triiodo, to monitor microvascular changes in tumors treated with the angiogenesis inhibitor bevacizumab. Ten female nude rats with MDA-MB 435 xenograft tumors were treated with 1 mg intraperitoneal bevacizumab when tumors reached 1 cm diameter and, for 4 rats, treated again 7 days later. Just before and 24 hours after the first injection of anti-VEGF antibody, the tumors were imaged by dynamic CT scans enhanced with PEG12000-Gen4-triiodo (n = 3 rats) or iohexol (n = 3 rats). The other 4 rats underwent dynamic CT scans enhanced with PEG12000-Gen4-triiodo just before and 24 hours after the second injection of anti-VEGF antibody. Microvascular leakiness (K(PS)) was calculated for the tumors using a 2-compartment tissue model. PEG12000-Gen4-triiodo-enhanced CT scans showed progressive reductions in K(PS) from day 1 to 2 to 9 (from 2.55 to 1.27 to 0.69 microL min(-1) cm(-3), respectively, P < 0.005 for each comparison of day 1-2 and day 2-9). No significant difference was seen in the K(PS) estimates derived from iohexol-enhanced CT scans obtained before or after treatment (276 vs. 223.8 microL min(-1) cm(-3), respectively, P = 0.54). The microvascular leak (K(PS)) was significantly larger for iohexol than for PEG12000-Gen4-triiodo-enhanced CT, P < 0.05. Dynamic macromolecular contrast-enhanced CT can be used to monitor serial decreases in tumor microvessel leakiness induced by repeated doses of an angiogenesis inhibitor drug.

A methodology for improving throughput using portal monitors

The National Internal Radiation Assessment Section (NIRAS), which operates the Canadian National Calibration Reference Centre for Bioassay and In Vivo Monitoring, has field deployable equipment for emergency response. A substantial part of this tool kit is a set of portal monitors that can be used to quickly screen people into the 'uncontaminated' and the 'contaminated'. The former term refers to a person who has <60 kBq (empirical practical detection limit) of activation/fission products and the latter group is contaminated by that amount or more. Recent field work has shown that one type of the NIRAS's portal monitors can be alarmed at significant distances if the level of contamination is high enough. The other types, which do not initiate a count until either an infra-red beam is broken or a proximity detector is activated, do not alarm but their background will be raised and this causes other problems. This paper proposes a method of group monitoring to help speed up the process of screening a large number of potentially contaminated persons using portal monitors. In short, the group of potentially contaminated persons will be kept isolated from the portal stations. Depending on a real-time estimate of the percentage of contaminated persons in the crowd, groups of persons will be selected for screening. The hypergeometric distribution has been used to decide on the sampling group size with an expectation that 90% of the time no contaminated person will be present in the group. Once removed from the main waiting area, the group will be pre-screened and then, depending on the result, sent to the appropriate portal. It is anticipated that this will greatly speed up processing as it substantially reduces the transit time. Transits times have also been estimated in addition to the number of personnel required to run all of NIRAS's field deployable equipment.

In Vivo Monitoring of the Prion Replication Cycle Reveals a Critical Role for Sis1 in Delivering Substrates to Hsp104

Prions in Saccharomyces cerevisiae are inherited ordered aggregates reliant upon the disaggregase Hsp104 for stable maintenance. The function of other factors in the natural prion cycle is unclear. We constructed yeast-bacterial chimeric chaperones to resolve the roles of Hsp104 domains, and by extension chaperones that interact with these domains, in prion propagation. Our results show that, as with amorphous aggregate dissolution, the Hsp70/40 system recruits prion substrates to Hsp104 via its top ring. By adapting our chimera to couple to an inactive protease "trap," we monitored the reaction products of prion propagation in vivo. We find that prion maintenance is accompanied by translocation of prion proteins through Hsp104 hexamers and that both processes critically rely upon the Hsp40 Sis1. Our data suggest that yeast prion replication is a natural extension of chaperone activity in dissolving amorphous aggregates, distinguished from its ancestral reaction by the ordered, self-propagating structure of the substrate.

In Vivo Monitoring of Focused Ultrasound Surgery Using Local Harmonic Motion

The present study established the feasibility of a technique for monitoring focused ultrasound (FUS) lesion formation in vivo using localized harmonic motion (LHM) measurements. Oscillatory motion (frequencies between 50 and 300 Hz) was generated within tissues by induction of a periodic radiation force with a FUS transducer. The harmonic motion was estimated using cross correlation of RF ultrasonic signals acquired at different instances during the motion by using a confocal diagnostic ultrasound transducer. The technique was evaluated in vivo in rabbit muscle (14 locations) in an magnetic resonance (MR) imager for simultaneous ultrasound harmonic motion tracking and MR thermometry. The measured maximum amplitude of the induced harmonic motion before and after the lesion formation was significantly different for all the tested motion frequencies, and decreased between 17 and 81% depending on the frequency and location. During the FUS exposure a drop in the maximum amplitude value was observed and a threshold value could be associated to the formation of a thermal lesion. A series of controlled sonications was performed by stopping the exposure when the threshold value in LHM amplitude was reached and the presence of a thermal lesion was confirmed by MR imaging. LHM measurements were also used to perform a spatial scan of the tissues across the exposure region and the thermal lesions could be detected as a reduction in the maximum motion amplitude value at the sonication region. ( E-mail: laura.curiel@sri.utoronto.ca)

Online In-Tube Solid-Phase Extraction Using a Nonfunctionalized PVC Tube Coupled with ICPMS for in Vivo Monitoring of Trace Metals in Rat Brain Microdialysates

We have developed a simple, automatic, online in-tube solid-phase extraction (SPE) method for the preconcentration of trace elements from saline samples. The method takes advantage of the adsorption of trace metal ions onto the interior of a nonfunctionalized PVC tube as a means of separation from the matrix salt. The adsorption of transition metal ions is presumably dominated by Lewis acid/base interactions, which facilitate the formation of metal-PVC complexes. For simultaneous determination of multiple trace metals in the extracellular fluid from the brains of anesthetized rats, we developed an online analytical system comprising microdialysis sampling, the established in-tube extraction procedure, and ICPMS. In the extraction step, the efficiency was optimal when the pH of the sample was adjusted to 8.0 using phosphate buffer. After extraction onto the interior of PVC tube, the adsorbed analytes were eluted with 0.5% HNO 3 prior to online ICPMS measurement. For those elements present in the extracellular fluid at less than nanogram-per-milliliter concentrations (i.e., Cu, Zn, and Mn), a temporal resolution of 12 min was required to collect enough microdialysate to meet the sensitivity requirements of the ICPMS instrument. Noteworthily, because washing and postconditioning the interior of PVC tube are not necessary, a relatively unsophisticated and clean procedure was attained and extremely low detection limits in the range of 5.0 to 510 ng L (-1) were thus obtained for the analysis of Mn, Cu, Zn, Pb, Cd, Ni, and Co in microdialysate samples of 8 microL by ICPMS. To the best of our knowledge, this study is the first to exploit PVC peristaltic tubing as an SPE adsorbent in the laboratory-on-valve mode for online sample treatment and trace metal preconcentration prior to ICPMS measurement. We confirmed the analytical reliability of this method through the analysis of the certified reference material SLEW-3 (estuarine water) and 2670a (human urine) and demonstrated its applicability through simultaneous in vivo monitoring of the basal concentrations and concentration profiles of multiple metal ions in the brain extracellular fluid of a living rat.

In Vivo Monitoring of Intravenously Injected Gold Nanorods Using Near‐Infrared Light

Gold nanorods showing surface plasmon (SP) bands in the near-IR region are used as bioimaging probes that respond to near-IR light in mice. The SP bands of intravenously injected polyethylene glycol-modified gold nanorods are directly monitored from the mouse abdomen by using a spectrophotometer equipped with an integrating sphere. The absorbance at 900 nm from the gold nanorods immediately increases after injection and reaches a plateau. The injection of phosphatidylcholine-modified gold nanorods also increases the absorbance at 900 nm, but the absorbance decreases single exponentially with a 1.3-min half-life. In vivo spectral changes of gold nanorods depend on the surface characteristics, and can be observed in real time using simple spectroscopic measurements.

In Vivo Monitoring of Inflammation After Cardiac and Cerebral Ischemia by Fluorine Magnetic Resonance Imaging

In this study, we developed and validated a new approach for in vivo visualization of inflammatory processes by magnetic resonance imaging using biochemically inert nanoemulsions of perfluorocarbons (PFCs). Local inflammation was provoked in 2 separate murine models of acute cardiac and cerebral ischemia, followed by intravenous injection of PFCs. Simultaneous acquisition of morphologically matching proton ((1)H) and fluorine ((19)F) images enabled an exact anatomic localization of PFCs after application. Repetitive (1)H/(19)F magnetic resonance imaging at 9.4 T revealed a time-dependent infiltration of injected PFCs into the border zone of infarcted areas in both injury models, and histology demonstrated a colocalization of PFCs with cells of the monocyte/macrophage system. We regularly found the accumulation of PFCs in lymph nodes. Using rhodamine-labeled PFCs, we identified circulating monocytes/macrophages as the main cell fraction taking up injected nanoparticles. PFCs can serve as a "positive" contrast agent for the detection of inflammation by magnetic resonance imaging, permitting a spatial resolution close to the anatomic (1)H image and an excellent degree of specificity resulting from the lack of any (19)F background. Because PFCs are nontoxic, this approach may have a broad application in the imaging and diagnosis of numerous inflammatory disease states.

Continuous In Vivo Monitoring of a Flexible Subcutaneous Sensor in Freely Moving Diabetic Rats

Subcutaneous tissue is frequently the target site for placement of continuous, real-time metabolic sensors. Since the 1960s, numerous research groups have developed needle-like sensor designs, patterned after the Clarke Electrode, to monitor glucose in subcutaneous tissue. These designs perform well in vitro but often fail in vivo due to sensor instability and tissue response. None of these studies focused on the mechanical properties of implanted sensors and how these properties may affect in vivo performance. To investigate the role of sensor stiffness on short term functionality we developed a low stiffness subcutaneous sensor patterned after the Clarke Electrode and tested it in rodents. The purpose of this study was two-fold. The first goal was to demonstrate the in vivo functionality of the flexible sensor. The second goal was to evaluate the effect of stiffness on functionality by co-implanting stiff and flexible sensors. In the first series of studies the low stiffness sensors provided glucose level measurements that fell within the A and B regions of the Clarke Error Grid 93.0% of the time. The results of the second study yielded similar accuracy; however, no performance difference was seen between the stiff and flexible sensors. We concluded that the flexible sensor works for at least 3days after implantation in the subcutaneous tissue of freely moving rats and that the key property of low stiffness has no differential effect on the accuracy of the sensor in the freely moving rodent model of these studies.

196. In Vivo Monitoring of a Conditional Replicating Adenovirus in Breast Cancer by pIX – Red Fluorescent Protein Capsid Labeling

196. In Vivo Monitoring of a Conditional Replicating Adenovirus in Breast Cancer by pIX – Red Fluorescent Protein Capsid Labeling

In Vivo Monitoring of Circadian Timing in Freely Moving Mice

In mammals, the principal circadian pacemaker driving daily physiology and behavioral rhythms is located in the suprachiasmatic nucleus (SCN) in the anterior hypothalamus. The neural output of SCN is essential for the circadian regulation of behavioral activity. Although remarkable progress has been made in revealing the molecular basis of circadian rhythm generation within the SCN, the output pathways by which the SCN exert control over circadian rhythms are not well understood. Most SCN efferents target the subparaventricular zone (SPZ), which resides just dorsal to the SCN. This output pathway has been proposed as a major component involved in the outflow for circadian regulation. We have examined the downstream pathway of the central clock by means of multiunit neural activity (MUA) in freely moving mice. SCN neural activity is tightly coupled to environmental photic input and anticorrelated with MUA rhythm in the SPZ. In Clock mutant mice exhibiting attenuated circadian locomotor rhythmicity, MUA rhythmicity in the SCN and SPZ is similarly blunted. These results suggest that the SPZ plays a functional role in relaying circadian and photic signals to centers involved in generating behavioral activity.

In Vivo Monitoring of Multiple Trace Metals in the Brain Extracellular Fluid of Anesthetized Rats by Microdialysis−Membrane Desalter−ICPMS

We have developed an on-line analytical system involving microdialysis (MD) sampling, a carbohydrate membrane desalter (CMD), and an inductively coupled plasma mass spectrometer (ICPMS) system for the simultaneous determination of multiple trace metals in the extracellular fluid (ECF) in the brains of anesthetized rats. The microdialysate that perfused from the animal at a flow rate of 0.5 microL/min was on-line transferred to the CMD to remove the high-sodium matrix, followed by ICPMS measurement. The role of the CMD in this on-line system was investigated in detail. With prior addition of EDTA to the microdialysate to form anionic complexes of the metal analytes and the use of NH4Cl as a regenerant to exchange Na(+) with NH4(+) ions, both quantitative recovery of the trace metal analytes and quantitative removal of the sodium matrix could be achieved. Two experimental modes of the monitoring system were constructed. For those metals (e.g., Cu, Zn, and Mn) that existed at (sub)nanogram-per-milliliter concentrations in the microdialysate, the temporal resolution was 10 min when using a 10 microL loop for sample collection, followed by CMD and ICPMS; for those elements (e.g., Ca and Mg) that existed at microgram-per-milliliter levels (or greater), near-real-time analysis was possible because the microdialysate could be led, bypassing the sample loop, directly to the CMD for desalting without any time delay. Further improvement of the temporal resolution for the low-concentration elements was not possible without decreasing the detection limits of mass detection. Among the eight trace metals tested using this on-line system, the method detection limits for Cu, Zn, Mn, Co, Ni, and Pb reached subnanogram-per-milliliter levels; for electrolyte species such as Ca and Mg, the detection limits were in the range of 50-100 ng/mL. Analytical accuracy, expressed as spike recovery, was 100% +/- 15% for all of the elements tested. We demonstrate the applicability of the proposed system through the successful measurement of the basal values of Ca, Mg, Cu, Zn, and Mn in the ECF of a living rat brain and through in vivo monitoring of the concentration profiles of Mn and Pt in the ECF after the injection of drugs (MnCl2 and cisplatin) into the rats. This microdialysis system is the first to offer real-time, in vivo monitoring of trace elements such as Ca and Mg.