Development of imaging mass spectrometry (IMS) dataset extractor software,IMS convolution

Imaging mass spectrometry (IMS) is a powerful tool for detecting and visualizing biomolecules in tissue sections. The technology has been applied to several fields, and many researchers have started to apply it to pathological samples. However, it is very difficult for inexperienced users to extract meaningful signals from enormous IMS datasets, and the procedure is time-consuming. We have developed software, called IMS Convolution with regions of interest (ROI), to automatically extract meaningful signals from IMS datasets. The processing is based on the detection of common peaks within the ordered area in the IMS dataset. In this study, the IMS dataset from a mouse eyeball section was acquired by a mass microscope that we recently developed, and the peaks extracted by manual and automatic procedures were compared. The manual procedure extracted 16 peaks with higher intensity in mass spectra averaged in whole measurement points. On the other hand, the automatic procedure using IMS Convolution easily and equally extracted peaks without any effort. Moreover, the use of ROIs with IMS Convolution enabled us to extract the peak on each ROI area, and all of the 16 ion images on mouse eyeball tissue were from phosphatidylcholine species. Therefore, we believe that IMS Convolution with ROIs could automatically extract the meaningful peaks from large-volume IMS datasets for inexperienced users as well as for researchers who have performed the analysis.     

Preparation and Function of Poly(acrylic acid)s Modified by Supramolecular Complex Composed of Porphinatoiron and a Cyclodextrin Dimer That Bind Diatomic Molecules (O2 and CO) in Aqueous Solution

Poly(acrylic acid) (PAA) is modified by 5‐(4‐β‐alanylaminophenyl)‐10,15,20‐tris(4‐sulfonatophenyl) porphinatoiron(III) to yield iron porphyrin‐bearing PAAs (FeP(n)s) through a condensation reaction. FeP(n)s were further functionalized by Py3CD, which is a per‐O‐methylated β‐cyclodextrin (CD) dimer with a pyridine linker and includes the porphyrin pendants to form ferric hemoCD‐P(n)s. Ferrous hemoCD‐P(3), having three porphyrin chromophores in a polymer chain, is shown to bind molecular oxygen (P_1/2=7.9±1.4 Torr) in aqueous solution at pH 7.0 and 25 °C, affording oxy‐hemoCD‐P(3). Oxy‐hemoCD‐P(3) is biphasically autoxidized to ferric hemoCD‐P(3), with 27 % of the dioxygen adducts being rapidly oxidized. The rate of autoxidation of oxy‐hemoCD‐P(15), having 15 porphyrin chromophores in a polymer chain, was much faster than that of oxy‐hemoCD‐P(3), thus suggesting self‐catalyzed autoxidation of oxy‐hemoCD‐P(n)s. Oxy‐hemoCD‐P(n)s are markedly stabilized by catalase, thereby indicating that hydrogen peroxide generated from oxy‐hemoCD‐P(n) accelerates the autoxidation. Most of the hemoCD‐P(3) molecules injected into the femoral vein of a rat remained in the body, though about 16 % of the hemoCD‐P(3) molecules were excreted in the urine as a carbon monoxide adduct.     

3D microfluidic chips with integrated functional microelements fabricated by a femtosecond laser for studying the gliding mechanism of cyanobacteria

Phormidium, a genus of filamentous cyanobacteria, forms endosymbiotic associations with seedling roots that accelerate the growth of the vegetable seedlings. Understanding the gliding mechanism of Phormidium will facilitate improved formation of this association and increased vegetable production. To observe the gliding movements, we fabricated various microfluidic chips termed nanoaquariums using a femtosecond (fs) laser. Direct fs laser writing, followed by annealing and successive wet etching in dilute hydrofluoric acid solution, can easily produce three-dimensional (3D) microfluidics with different structures embedded in a photostructurable glass. Using the fs laser, optical waveguides and filters were integrated with the microfluidic structures in the microchips, allowing the gliding mechanism to be more easily clarified. Using this apparatus, we found that CO(2) secreted from the seedling root attracts Phormidium in the presence of light, and determined the light intensity and specific wavelength necessary for gliding.     

Measurements on diproton emission from the break-up channels of 23Al and 22Mg

Two-proton relative momentum distributions from the break-up channels ²³Al→p+p+²¹Na and ²²Mg→p+p+²⁰Ne at an energy of 60–70 A MeV have been measured together with two-proton opening angles at the projectile fragment separator beamline (RIPS) in the RIKEN Ring Cyclotron Facility. The results demonstrate the existence of diproton emission component from single-step ²He for highly excited ²³Al and ²²Mg.     

Chilling out: a cool aqueous environment promotes the formation of gas-surface complexes

SO(2), an important atmospheric pollutant, has been implicated in environmental phenomena such as acid rain, climate change, and cloud formation. In addition, SO(2) is fundamentally interesting because it forms spectroscopically identifiable complexes with water at aqueous surfaces. Vibrational sum frequency spectroscopy (VSFS) is used here to further investigate the mechanism by which SO(2) adsorbs to water at tropospherically relevant temperatures (0-23 °C). The spectral results lead to two important conclusions. SO(2) surface affinity is enhanced at colder temperatures, with nearly all of the topmost water molecules showing evidence of binding to SO(2) at 0 °C as compared to a much lower fraction at room temperature. This surface adsorption results in significant changes in water orientation at the surface, but is reversible at the temperatures examined here. Second, the SO(2) complex formation at aqueous surfaces is independent of aqueous solution acidity. One challenge in previous uptake studies was the ability to distinguish between the effects of surface adsorption as compared to bulk accommodation. The surface and vibrational specificity of these studies make this distinction possible, allowing a selective study of how the aqueous properties temperature and pH influence SO(2) surface affinity.     

Enantiomeric separation of asymmetric triacylglycerol by recycle high-performance liquid chromatography with chiral column

In our previous studies, we employed recycle HPLC for the separation of triacylglycerol (TAG)-positional isomers (PIs). In this study, a recycle HPLC system equipped with a polysaccharide-based chiral column was applied to the enantiomeric separation of some asymmetric TAGs having straight-chain C16-C18 acyl residues. As a result, 1,2-dipalmitoyl-3-oleoyl-rac-glycerol (rac-PPO), 1,2-dioleoyl-3-palmitoyl-rac-glycerol (rac-OOP), and 1,2-dipalmitoyl-3-linoleoyl-rac-glycerol (rac-PPL) were resolved into their respective enantiomers. However, neither 1,2-dioleoyl-3-linoleoyl-rac-glycerol (rac-OOL), consisting of only unsaturated fatty acids, nor 1,2-dipalmitoyl-3-stearoyl-rac-glycerol (rac-PPS), consisting of only saturated fatty acids, was resolved. These results suggest that the asymmetric TAGs, used in this study, having both a palmitic acid moiety and an oleic acid (or a linoleic acid) moiety at the sn-1 or sn-3 positions are resolved by the chiral column. This new chiral separation method can be used in combination with atmospheric pressure chemical ionization mass spectrometry to determine the sn-OOP/sn-POO ratio in palm oil. This method is applicable for the chiral separation of asymmetric TAGs in palm oil.     

Size separation of colloidally dispersed nanoparticles using a monolithic capillary column

We developed a method to separate colloidally dispersed nanoparticles on monolithic capillary columns. Silica nanoparticles were eluted according to their sizes, and the plots of the logarithm of the size of nanoparticles against their elution volume showed good linearity (r=0.992) over wide range of sizes. Because of the high porosity of the monolithic column (porosity; 88%), the column's length could be increased without clogging of the dispersed samples and the pressure in a long column (500 mm × 0.2 mm i.d.) was low, with a value of 5.8 MPa at a flow rate of 1 μL/min. We demonstrate that this method using monolithic capillary columns could be used as a powerful tool for size separation of nanometer-size materials, which will open a new pathway to quality control of nanomaterials in nanotechnology applications.     

Efficient biotinylation of methacryloyl-functionalized nonadherent cells for formation of cell microarrays

In order to obtain cell microarrays formed with human promyelocytic leukemia cells (HL-60), biotinylation of methacryloyl-functionalized HL-60 cells was performed via a thiol-ene reaction with thiol-terminated 4-arm poly(ethylene glycol) (PEG(4)10K-SH). Biotinylated HL-60 cells were selectively adhered onto an avidin-patterned surface with high viability.