The tumor affinity of chlorin e6 and its sonodynamic effects on non-small cell lung cancer

ObjectiveSonodynamic therapy (SDT) is a promising new approach for cancer therapy. The aim of this study was to investigate the tumor affinity of chlorin e6, a photosensitizer, and its sonodynamic effects on NSCLC.MethodsHuman lung adenocarcinoma cells SPCA-1 and mice bearing SPCA-1 tumor xenograft were exposed to ultrasound in the presence or absence of chlorin e6. Chlorin e6 distribution was detected by laser scan confocal microscope. Cell apoptosis and necrosis were studied by flow cytometry analysis. Tumor size and weight were measured after different treatments.ResultsThe concentration of chlorin e6 in tumor tissue was remarkably higher than that in normal muscle near tumor, and the difference was greatest at 18 h (the fluorescence intensity was 5.38-fold higher in tumor than in muscle, P < 0.05). In vivo, ultrasound (0.4–1.6 W/cm²) or chlorin e6 (10–40 mg/kg) alone had no remarkable anti-tumor effects, but the combination of ultrasound (1.6 W/cm²) with chlorin e6 (SDT) hampered tumor growth significantly (P < 0.05). Intraperitoneal injection of 40 mg/kg chlorin e6 exerted no notable side effect on blood, liver and kidney function. Flow cytometry analysis showed that chlorin e6-mediated sonodynamic effect was mainly through the induction of cell necrosis.ConclusionChlorin e6 is a promising sonosensitizer and chlorin e6-mediated SDT may provide a new approach for NSCLC therapy.     

Relative quantification of cellular sections with molecular depth profiling ToF-SIMS imaging

We report the use of secondary ion mass spectrometry (SIMS) imaging to quantify the relative difference in the amount of lipid between two sections, the plasma membrane and the cytoplasm, of single cells from two different populations. Cells were each labeled with lipophillic dyes, frozen, fractured and analyzed in a ToF-SIMS mass spectrometer equipped with a 40 keV C₆₀⁺ ion source. In addition to identifying cells from separate populations, the lipophilic dyes can be used as a marker for the outer leaflet of the cell membrane and therefore as a depth finder. Here, we show that it is possible to compare the amount of lipids with particular headgroups in the cell membrane of a treated cell to the membrane of a control cell. Following erosion of the cell membranes, the amount of the two specific lipid head groups in the cytoplasm of the treated cell can be compared to those lipids in a control cell. Here we take the first step in this experimental design and display the ability to analyze multiple sections of frozen cells following a single fracture.     

Reliable measurement of the FRET sensitized-quenching transition factor for FRET quantification in living cells

3-cube-based Förster resonance energy transfer (FRET) microscopy, a sensitized acceptor FRET quantification method, has been widely used to visualize dynamic protein–protein interaction in living cells. Determining the FRET sensitized-quenching transition factor (G factor) of a particular donor-acceptor pair and optical system is crucial for 3-cube FRET quantification. We here improved the acceptor photobleaching-based G factor determination method (termed as mPb-G) and the two-plasmid-based G factor determination method (termed as mTP-G) for rapid and reliable measurement of the G factor. mTP-G method determines G factor by simultaneously detecting three images of cells exclusively expressing each of two tandem constructs with multiple donors and multiple acceptors. This method circumvents switchover of the cells exclusively expressing each of the two constructs. mPb-G method images G factor by detecting three images of cells expressing a donor-acceptor tandem FRET construct before and after partially photobleaching acceptor. We performed the two methods on our dual-channel wide-field FRET microscope to obtain reliable G factor, and also measured the FRET efficiency and acceptor-to-donor concentration ratio of tandem constructs with different acceptor-donor stoichiometries in living HepG2 cells. mTP-G and mPb-G methods provide two simple and reliable tools for determining the G factor, in turn, quantitatively measuring FRET signal and monitoring dynamic biochemical processes in living cells.     

Structural and permeability sensitivity of cells to low intensity ultrasound: Infrared and fluorescence evidence in vitro

This work is focused on the in vitro study of the effects induced by medical ultrasound (US) in murine fibroblast cells (NIH-3T3) at a low-intensity of exposure (spatial peak temporal average intensity I_ta < 0.1 W cm⁻²). Conventional 1 MHz and 3 MHz US devices of therapeutic relevance were employed with varying intensity and exposure time parameters. In this framework, upon cells exposure to US, structural changes at the molecular level were evaluated by infrared spectroscopy; alterations in plasma membrane permeability were monitored in terms of uptake efficiency of small cell-impermeable model drug molecules, as measured by fluorescence microscopy and flow cytometry. The results were related to the cell viability and combined with the statistical PCA analysis, confirming that NIH-3T3 cells are sensitive to therapeutic US, mainly at 1 MHz, with time-dependent increases in both efficiency of uptake, recovery of wild-type membrane permeability, and the size of molecules entering 3T3. On the contrary, the exposures from US equipment at 3 MHz show uptakes comparable with untreated samples.     

Sonodynamic action of pyropheophorbide-a methyl ester induces mitochondrial damage in liver cancer cells

Sonodynamic therapy with pyropheophorbide-a methyl ester (MPPa) presents a promising aspect in treating liver cancer. The present study aims to investigate the mitochondrial damage of liver cancer cells induced by MPPa-mediated sonodynamic action. Mouse hepatoma cell line H₂₂ cells were incubated with MPPa (2 μM) for 20 h and then exposed to ultrasound with an intensity of 0.97 W/cm² for 8 s. Cytotoxicity was investigated 24 h after sonodynamic action using MTT assay and light microscopy. Mitochondrial membrane potential (ΔΨm) was analyzed using flow cytometry with rhodamine 123 staining and ultrastructural changes were observed using transmission electron microscopy (TEM).The cytotoxicity of MPPa-mediated SDT on H₂₂ cell line was 73.00 ± 3.42%, greater than ultrasound treatment alone (28.12 ± 5.19%) significantly while MPPa treatment alone had no significant effect on H₂₂ cells. Moreover, after MPPa-mediated SDT cancer cells showed swollen mitochondria under TEM and a significant collapse of mitochondrial membrane potential. Our findings demonstrated that MPPa-mediated SDT could remarkably induce cell death of H₂₂ cells, and highlighted that mitochondrial damage might be an important cause of cell death induced by MPPa-mediated SDT.     

Antibacterial mechanism of ultrasound against Escherichia coli: Alterations in membrane microstructures and properties

This study was aimed at providing new insights on the response of bacterial cell membranes to ultrasound exposure. Escherichia coli (E. coli) O157:H7 cells were exposed to different ultrasound treatments (power intensities of 64, 191, 372, and 573 W/cm², frequency of 20 kHz, pulsed mode of 2 sec: 2 sec) and the dynamic changes in cell viability within 27 min were assessed. With an increase in ultrasonic intensity and prolonged duration, a 0.76–3.52 log CFU/mL reduction in E. coli populations was attained. The alterations in the sensitivity of ultrasound-treated cells to antimicrobial compounds were evaluated by exposure to thyme essential oil nanoemulsion (TEON). The treatment reduced the E. coli population by 2.16–7.10 log CFU/mL, indicating the effects of ultrasonic field on facilitating the antibacterial efficacy of TEON. Ultrasonic-treated E. coli cells also displayed remarkable morphological and ultrastructural damages with destroyed membrane integrity and misshaped cell structures, which was observed by electron microscopy analysis. Significant increase in outer and inner membrane permeability, along with the cytoplasmic leakage and membrane depolarization were assessed utilizing spectrophotometry. For the first time, significant reduction in the membrane fluidity in response to ultrasound exposure were investigated. Additional efforts in exploring the effect of ultrasonic field on some bacterial membrane compositions were performed with infrared spectroscopy. In this study, multiple lines of evidence effectively served to elucidate the alterations on cellular membrane structure and property during exposure to sonication that could extend our understanding of the antimicrobial molecular mechanisms of ultrasound.     

Ultrasound velocimetry of ferrofluid spin-up flow measurements using a spherical coil assembly to impose a uniform rotating magnetic field

Ferrofluid spin-up flow is studied within a sphere subjected to a uniform rotating magnetic field from two surrounding spherical coils carrying sinusoidally varying currents at right angles and 90° phase difference. Ultrasound velocimetry measurements in a full sphere of ferrofluid shows no measureable flow. There is significant bulk flow in a partially filled sphere (1-14 mm/s) of ferrofluid or a finite height cylinder of ferrofluid with no cover (1-4 mm/s) placed in the spherical coil apparatus. The flow is due to free surface effects and the non-uniform magnetic field associated with the shape demagnetizing effects. Flow is also observed in the fully filled ferrofluid sphere (1-20 mm/s) when the field is made non-uniform by adding a permanent magnet or a DC or AC excited small solenoidal coil. This confirms that a non-uniform magnetic field or a non-uniform distribution of magnetization due to a non-uniform magnetic field are causes of spin-up flow in ferrofluids with no free surface, while tangential magnetic surface stress contributes to flow in the presence of a free surface.Recent work has fitted velocity flow measurements of ferrofluid filled finite height cylinders with no free surface, subjected to uniform rotating magnetic fields, neglecting the container shape effects which cause non-uniform demagnetizing fields, and resulting in much larger non-physical effective values of spin viscosity η′∼10⁻⁸−10⁻¹² N s than those obtained from theoretical spin diffusion analysis where η′≤10⁻¹⁸ N s. COMSOL Multiphysics finite element computer simulations of spherical geometry in a uniform rotating magnetic field using non-physically large experimental fit values of spin viscosity η′∼10⁻⁸−10⁻¹² N s with a zero spin-velocity boundary condition at the outer wall predicts measureable flow, while simulations setting spin viscosity to zero (η′=0) results in negligible flow, in agreement with the ultrasound velocimetry measurements. COMSOL simulations also confirm that a non-uniform rotating magnetic field or a uniform rotating magnetic field with a non-uniform distribution of magnetization due to an external magnet or a current carrying coil can drive a measureable flow in an infinitely long ferrofluid cylinder with zero spin viscosity (η′=0).     

In vivo monitoring of multiple circulating cell populations using two-photon flow cytometry

To detect and quantify multiple distinct populations of cells circulating simultaneously in the blood of living animals, we developed a novel optical system for two-channel, two-photon flow cytometry in vivo. We used this system to investigate the circulation dynamics in live animals of breast cancer cells with low (MCF-7) and high (MDA-MB-435) metastatic potential, showing for the first time that two different populations of circulating cells can be quantified simultaneously in the vasculature of a single live mouse. We also non-invasively monitored a population of labeled, circulating red blood cells for more than two weeks, demonstrating that this technique can also quantify the dynamics of abundant cells in the vascular system for prolonged periods of time. These data are the first in vivo application of multichannel flow cytometry utilizing two-photon excitation, which will greatly enhance our capability to study circulating cells in cancer and other disease processes.     

Quantitative MR imaging of targeted SPIO particles on the cell surface and comparison to flow cytometry

Purpose

To detect anti-CEACAM5 targeted superparamagnetic iron oxide (SPIO) particles in vitro on the cell surface by quantitative magnetic resonance (MR) imaging and to compare with flow cytometry.

Materials and Methods

The monoclonal mouse antibody T84.1 and an appropriate IgG isotype antibody were conjugated to dextran-coated SPIO particles. HT29 cells expressing carcinoembryonic antigen (CEACAM5) were treated with antibody-conjugated SPIO particles. Purified cell samples were examined on a 3.0-T MR scanner using a multi-echo spin-echo sequence for MR relaxometry. Aliquots of the cell samples were further treated with a fluorescein isothiocyanate (FITC) anti-dextran antibody and an Alexa Fluor 488 anti-mouse antibody for the corresponding flow cytometry.

Results

MR relaxometry revealed a dose-dependent binding of T84.1-conjugated SPIO particles with a positive correlation between R₂ relaxation rate of cell samples and SPIO particle concentration during incubation (r=0.993, P<.01). Positive correlations were also observed between R₂ relaxation rate and flow cytometry (geometric mean) with both fluorescent antibodies (r=0.972 and r=0.953, both P<.01), respectively.

Conclusion

The study revealed the feasibility of quantitative MR imaging of targeted SPIO particles on the cell surface comparable to flow cytometry.     

Characterization of drug-eluting stent (DES) materials with cluster secondary ion mass spectrometry (SIMS)

Secondary ion mass spectrometry (SIMS) employing an SF₅⁺ polyatomic primary ion source was utilized to analyze several materials commonly used in drug-eluting stents (DES). Poly(ethylene-co-vinyl acetate) (PEVA), poly(lactic-co-glycolic acid) (PLGA) and various poly(urethanes) were successfully depth profiled using SF₅⁺ bombardment. The resultant molecular depth profiles obtained from these polymeric films showed very little degradation in molecular signal as a function of increasing SF₅⁺ primary ion dose when experiments were performed at low temperatures (signal was maintained for doses up to ∼5 × 10¹⁵ ions/cm²). Temperature was determined to be an important parameter in both the success of the depth profiles and the mass spectral analysis of the polymers. In addition to the pristine polymer films, paclitaxel (drug released in Taxus™ stent) containing PLGA films were also characterized, where it was confirmed that both drug and polymer signals could be monitored as a function of depth at lower paclitaxel concentrations (10 wt%).     

Sensitivity Optimization in Continuous-Flow FTNMR

Equations simulating the steady-state magnetization of liquids in continuous-flow FTNMR are derived using a classical vector model, assuming plug flow. These equations are applied to calculation of (S/N)_t, the relative signal/noise per unit time of any nucleus undergoing any degree of Overhauser enhancement either in the detection cell or upstream, or both, and to optimization of experimental conditions, including pulse repetition timeT_rep, pulse angle β, and flow rate. Ideal parameters include a pulse angle of 90° and aT_repvalue equal to sample residence time in the NMR detection cell. Optimal flow rates are directly proportional to the premagnetization volume (the portion of sample equilibrated with the magnetic field prior to detection) and inversely proportional to spin–lattice relaxation timesT₁. Optimal premagnetization times are smaller than previously assumed, varying from about 1.1 to 1.9T₁values. (S/N)_tfor static FTNMR is discussed in some detail, and a new graphical method is presented for its optimization. Flow advantage, the (S/N)_tof optimized flow FTNMR experiments compared to that of static FTNMR in a given detection cell, is proportional to the square root of the ratio of premagnetization to detection cell volumes, and virtually independent of[formula]where[formula]is the apparent transverse-relaxation time. The theory is applied to examples from recent literature, including dynamic electron–nuclear polarization, and the literature is critically reviewed. The analysis shows that claims by previous authors of recycled flow FTNMR by itself leading to increased (S/N)_tfor slowly relaxing resonances are misleading, owing to underdetermination of (S/N)_tin static measurements and failure to account for greater sample sizes required in flow experiments. For monitoring and control of chemical processes, the theory presented here enables the first rational basis for the design of a flow FTNMR apparatus and for the selection of acquisition parameters.     

Hypocrellin B-mediated sonodynamic action induces apoptosis of hepatocellular carcinoma cells

Objective

The present study aims to investigate apoptosis of hepatocellular carcinoma cells induced by hypocrellin B-mediated sonodynamic action.

Methods

The hypocrellin B concentration was kept constant at 2.5 μM and cells from the hepatocellular carcinoma HepG2 cell line were exposed to ultrasound with an intensity of 0.46 W/cm² for 8 s. Cell cytotoxicity was quantified using an MTT assay 24 h after sonodynamic therapy (SDT) of hypocrellin B. Apoptosis was investigated using a flow cytometry with Annexin V-FITC and propidium iodine staining. Intracellular reactive oxygen species (ROS) levels were detected using a flow cytometry with 2,7-dichlorodihydrofluorecein diacetate (DCFH-DA) staining.

Results

The cytotoxicity of hypocrellin B-mediated sonodynamic action on HepG2 cells was significantly higher than those of other treatments including ultrasound alone, hypocrellin B alone and sham treatment. Flow cytometry showed that hypocrellin B-induced sonodynamic action markedly enhanced the apoptotic rate of HepG2 cells. Increased ROS was observed in HepG2 cells after being treated with hypocrellin B-mediated sonodynamic action.

Conclusions

Our data demonstrated that hypocrellin B-mediated sonodynamic action remarkably induced apoptosis of HepG2 cells, suggesting that apoptosis is an important mechanism of cell death induced by hypocrellin B-mediated SDT.     

4D Flow MRI hemodynamic benchmarking of surgical bioprosthetic valves

PurposeWe exploited 4-dimensional flow magnetic resonance imaging (4D Flow), combined with a standardized in vitro setting, to establish a comprehensive benchmark for the systematic hemodynamic comparison of surgical aortic bioprosthetic valves (BPVs).Materials and methods4D Flow analysis was performed on two small sizes of three commercialized pericardial BPVs (Trifecta™ GT, Carpentier-Edwards PERIMOUNT Magna and Crown PRT®). Each BPV was tested over a clinically pertinent range of continuous flow rates within an in vitro MRI-compatible system, equipped with pressure transducers. In-house 4D Flow post-processing of the post-valvular velocity field included the quantification of BPV effective orifice area (EOA), transvalvular pressure gradients (TPG), kinetic energy and viscous energy dissipation.ResultsThe 4D Flow technique effectively captured the 3-dimensional flow pattern of each device. Trifecta exhibited the lowest range of velocity and kinetic energy, maximized EOA (p < 0.0001) and minimized TPGs (p ≤ 0.015) if compared with Magna and Crown, these reporting minor EOA difference s (p ≥ 0.042) and similar TPGs (p ≥ 0.25). 4D Flow TPGs estimations strongly correlated against ground-truth data from pressure transducers; viscous energy dissipation proved to be inversely proportional to the fluid jet penetration.ConclusionThe proposed 4D Flow analysis pinpointed consistent hemodynamic differences among BPVs, highlighting the not negligible effect of device size on the fluidynamic outcomes. The efficacy of non-invasive 4D Flow MRI protocol could shed light on how standardize the comparison among devices in relation to their actual hemodynamic performances and improve current criteria for their selection.     

Decay time shortening of fluorescence from donor-acceptor pair proteins using ultrafast time-resolved fluorescence resonance energy transfer spectroscopy

We improved an ultrafast time-resolved fluorescence resonance energy transfer (FRET) spectroscopy system and measured directly the decrease in the fluorescence decay time of the FRET signal, without any entanglement of components in the picosecond time scale from the donor-acceptor protein pairs (such as cameleon protein for calcium ion indicator, and ligand-activated GRIN-Go proteins pair). The drastic decrease in lifetime of the donor protein fluorescence under the FRET condition (e.g. a 47.8% decrease for a GRIN-Go protein pair) proves the deformation dynamics between donor and acceptor fluorescent proteins in an activated state of a mixed donor-acceptor protein pair. This study is the first clear evidence of physical contact of the GRIN-Go proteins pair using time-resolved FRET system. G protein-coupled receptors (GPCRs) are the most important protein family for the recognition of many chemical substances at the cell surface. They are the targets of many drugs. Simultaneously, we were able to observe the time-resolved spectra of luminous proteins at the initial stage under the FRET condition, within 10 ns from excitation. This new FRET system allows us to trace the dynamics of the interaction between proteins at the ligand-induced activated state, molecular structure change and combination or dissociation. It will be a key technology for the development of protein chip technology.     

Photobleaching-Based Quantitative Analysis of Fluorescence Resonance Energy Transfer inside Single Living Cell

The current advances of fluorescence microscopy and new fluorescent probes make fluorescence resonance energy transfer (FRET) a powerful technique for studying protein-protein interactions inside living cells. It is very hard to quantitatively analyze FRET efficiency using intensity-based FRET imaging microscopy due to the presence of autofluorescence and spectral crosstalks. In this study, we for the first time developed a novel photobleaching-based method to quantitatively detect FRET efficiency (Pb-FRET) by selectively photobleaching acceptor. The Pb-FRET method requires two fluorescence detection channels: a donor channel (CH ₁ ) to selectively detect the fluorescence from donor, and a FRET channel (CH ₂ ) which normally includes the fluorescence from both acceptor and donor due to emission spectral crosstalk. We used the Pb-FRET method to quantitatively measure the FRET efficiency of SCAT3, a caspase-3 indicator based on FRET, inside single living cells stably expressing SCAT3 during STS-induced apoptosis. At 0, 6 and 12 h after STS treatment, the FRET efficiency of SCAT3 obtained by Pb-FRET inside living cells was verified by two-photon excitation (TPE) fluorescence lifetime imaging microscopy (FLIM). The temporal resolution of Pb-FRET method is in second time-scale for ROI photobleaching, even in microsecond time-scale for spot photobleaching. Our results demonstrate that the Pb-FRET method is independent of photobleaching degree, and is very useful for quantitatively monitoring protein-protein interactions inside single living cell.     

New insights on the role of ROS in the mechanisms of sonoporation-mediated gene delivery

Reactive oxygen species (ROS) are hypothesized to play a role in the sonoporation mechanisms. Nevertheless, the acoustical phenomenon behind the ROS production as well as the exact mechanisms of ROS action involved in the increased cell membrane permeability are still not fully understood. Therefore, we investigated the key processes occurring at the molecular level in and around microbubbles subjected to ultrasound using computational chemistry methods. To confirm the molecular simulation predictions, we measured the ROS production by exposing SonoVue® microbubbles (MBs) to ultrasound using biological assays. To investigate the role of ROS in cell membrane permeabilization, cells were subjected to ultrasound in presence of MBs and plasmid encoding reporter gene, and the transfection level was assessed using flow cytometry. The molecular simulations showed that under sonoporation conditions, ROS can form inside the MBs. These radicals could easily diffuse through the MB shell toward the surrounding aqueous phase and participate in the permeabilization of nearby cell membranes. Experimental data confirmed that MBs favor spontaneous formation of a host of free radicals where HO was the main ROS species after US exposure. The presence of ROS scavengers/inhibitors during the sonoporation process decreased both the production of ROS and the subsequent transfection level without significant loss of cell viability. In conclusion, the exposure of MBs to ultrasound might be the origin of chemical effects, which play a role in the cell membrane permeabilization and in the in vitro gene delivery when generated in its proximity.     

Molecular ion yield enhancement induced by gold deposition in static secondary ion mass spectrometry

Static ToF-SIMS was used to evaluate the effect of gold condensation as a sample treatment prior to analysis. The experiments were carried out with a model molecular layer (Triacontane M = 422.4 Da), upon atomic (In⁺) and polyatomic (Bi₃⁺) projectile bombardment. The results indicate that the effect of molecular ion yield improvement using gold metallization exists only under atomic projectile impact. While the quasi-molecular ion (M+Au)⁺ signal can become two orders of magnitude larger than that of the deprotonated molecular ion from the pristine sample under In⁺ bombardment, it barely reaches the initial intensity of (M−H)⁺ when Bi₃⁺ projectiles are used. The differences observed for mono- and polyatomic primary ion bombardment might be explained by differences in near-surface energy deposition, which influences the sputtering and ionization processes.     

Model multilayer structures for three-dimensional cell imaging

The prospects for SIMS three-dimensional analysis of biological materials were explored using model multilayer structures. The samples were analyzed in a ToF-SIMS spectrometer equipped with a 20 keV buckminsterfullerene (C₆₀⁺) ion source. Molecular depth information was acquired using a C₆₀⁺ ion beam to etch through the multilayer structures at specified time intervals. Subsequent to each individual erosion cycle, static SIMS spectra were recorded using a pulsed C₆₀⁺ ion probe. Molecular intensities in sequential mass spectra were monitored as a function of primary ion fluence. The resulting depth information was used to characterize C₆₀⁺ bombardment of biological materials. Specifically, molecular depth profile studies involving dehydrated dipalmitoyl-phosphatidylcholine (DPPC) organic films indicate that cell membrane lipid materials do not experience significant chemical damage when bombarded with C₆₀⁺ ion fluences greater than 10¹⁵ ions/cm². Moreover, depth profile analyses of DPPC-sucrose frozen multilayer structures suggest that biomolecule information can be uncovered after the C₆₀⁺ sputter removal of a 20 nm overlayer with no appreciable loss of underlying molecular signal. The experimental results support the potential for three-dimensional molecular mapping of biological materials using cluster SIMS.     

Ultrasound-mediated disruption of cell membranes. II. Heterogeneous effects on cells

Ultrasound has been shown to reversibly and irreversibly disrupt membranes of viable cells through a mechanism believed to involve cavitation. Because cavitation is both temporally and spatially heterogeneous, flow cytometry was used to identify and quantify heterogeneity in the effects of ultrasound on molecular uptake and cell viability on a cell-by-cell basis for suspensions of DU145 prostate cancer and aortic smooth muscle cells exposed to varying peak negative acoustic pressures (0.6-3.0 MPa). exposure times (120-2,000 ms), and pulse lengths (0.02-60 ms) in the presence of Optison (1.7% v/v) contrast agent. Cell-to-cell heterogeneity was observed at all conditions studied and was classified into three subpopulations: nominal uptake (NUP), low uptake (LUP), and high uptake (HUP) populations. The average number of molecules within each subpopulation was generally constant: 10(4)-10(5) molecules/cell in NUP, approximately 10(6) molecules/cell in LUP, and approximately 10(7) molecules/cell in HUP. However, the fraction of cells within each subpopulation showed a strong dependence on both acoustic pressure and exposure time. Varying pulse length produced no significant effect. The distribution of cells among the three subpopulations correlated with acoustic energy exposure, which suggests that energy exposure may govern the ability of ultrasound to induce bioeffects by a nonthermal mechanism.     

Dipolar and quadrupolar detection using an FT-ICR MS setup at the MPIK Heidelberg

Dipolar and single-phase two-electrode quadrupolar detection schemes have been investigated at a Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS) setup built for the KATRIN experiment at the Max-Planck-Institute for Nuclear Physics (MPIK) in Heidelberg. We present first experimental results of ⁷Li^?+? signals from a cylindrical Penning trap configuration for both detection schemes. While the prominent signal of the conventional dipolar detection scheme marks the reduced cyclotron frequency, the main signal for the quadrupolar detection appears at the sum of the reduced cyclotron frequency and the magnetron frequency. For ideal trapping fields, this sum frequency equals the ion cyclotron frequency ?? _c ?=?qB/(2??m). Sidebands due to the combined motions of the cyclotron mode and magnetron mode are observed by quadrupolar detection which allows the determination of the respective combinations of eigenfrequencies.