High-resolution Differential Mobility Research Articles

Opinion: Should high-resolution differential mobility analyzers be used in mainstream aerosol studies?

Abstract. Differential mobility analyzers (DMAs) are widely used instruments to measure the size distributions of submicron aerosols. High-resolution DMAs (HRDMAs) are defined here as plain DMAs maintaining a steady flow over an unusually broad range of sheath gas flow rates Q. HRDMAs, first developed by Georg Reischl's group, have existed for a long time. However, they have not been widely adopted, except in the size range below 10 nm, often in new particle formation studies. Here we question the commonly held view that HRDMAs are necessarily complex, bulky and expensive machines, mainly of interest in exotic applications outside mainstream aerosol research. Rather, many studies central to aerosol research could be carried out with HRDMAs with considerable advantage in size range, resolution, sensitivity and measurement speed. DMA manufacturers will hopefully take the challenge of developing commercial HRDMAs of complexity and cost comparable to those of today's commercial instruments, adapted for broad use by aerosol scientists, though with greatly improved flexibility and performance. Some of the technical challenges that still need to be overcome are discussed, such as the development of high-flow condensation counter detectors, and the control of high sample and sheath gas flow rates.

Technical note: Characterization of polystyrene latex spheres 300 nm in diameter with a singularly narrow size distribution

We have recently combined improved polystyrene latex (PSL) particle synthesis methods and new high resolution differential mobility analyzers (DMAs) covering the viral size range (up to about 300 nm) to demonstrate the availability of ∼200 nm PSL spheres with unusually narrow mobility distributions, with relative full width at half maximum (FWHM) as small as 1/60. By similar techniques we now report on the synthesis and analysis of PSL particles with diameters of about 300 nm achieving FWHM as small as 1/80. This is as narrow as the narrowest mobility peaks previously found with all but one of the viral particle so far studied with high resolution DMAs. The electrosprayed 300 nm particles produce two resolvable peaks, with the least mobile one being apparently an artifact of the electrospraying or charge reduction process. We conclude that future PSL synthesis improvements should be able to provide particles yielding mobility peaks with FWHM as small as 1/100.

A high resolution differential mobility analyzer for the study of large virus particles

The conical inner electrode of a recently described differential mobility analyzer (DMA), covering at resolution >50 the size range up to 60 nm, has been modified by increasing its radius at the outlet slit from 1 cm to 1.6 cm, and decreasing its convergence angle from 3° to 0.5°. The cylindrical outer electrode radius is maintained at 2 cm, while other critical dimensions are only slightly modified. The classification voltage for fixed flow rate of sheath gas and particle diameter decreases almost fourfold, extending the size range theoretically spanned at high resolution well beyond 120 nm. Tests with multiply charged polystyrene latex spheres 200 nm in diameter confirm the actual ability to cover this wide size range. A resolution above 100 (with no corrections for instrument or particle nonidealities) is demonstrated with rotavirus double-layered particles (DLP) whose mobility diameter is found to be near 60 nm. The DLP has several configurations that complicate its use for DMA characterization, but occasionally presents mainly one conformer, with singularly narrow relative mobility width FWHM < 1/150. Two broad range instruments with slightly different axial lengths have been developed and tested, both reaching resolving powers of 100.

Ions Generated from a Premixed Methane-Air Flame: Mobility Size Distributions and Charging Characteristics

ABSTRACT Chemical ionization in combustion systems forms concentrated ions of both polarities. Applying electric fields and plasmas to combustion systems has been shown to reduce emissions (such as particulate matter) potentially by controlling the ionic properties. Detailed flame-generated ion properties need to be characterized to better understand and predict the dynamics and roles of these ions in combustion and particle formation processes. In this work, we used a high-resolution differential mobility analyzer (HR-DMA) to map the mobility and size distributions of positive and negative ions generated from a premixed methane-air flat flame under atmospheric pressure. Measurements were conducted over a wide range of stoichiometric ratios (0.8 to 1.2) and heights above the burner (HAB, 7–42 mm). Positively charged ions are relatively stable over the entire range of experimental conditions, showing two major modes, one at 1.16 nm, and another at 1.42 nm. The mode corresponding to 1.42 nm showed a gradual increase in size with HAB, indicating the charging of hydrocarbon precursors. With the mobility values of the ions, we calculated their approximate mass values based on the empirical mobility–mass relationship and estimated the charging characteristics of particles in the flame. The ion profiles and particle charging characteristics obtained in this study will improve our understanding of electrostatic interactions in flame systems.

Singularly narrow size distributions of 200 nm polystyrene latex spheres determined by high resolution mobility analysis

A high-resolution differential mobility analyzer (DMA) is used for the first time to study the mobility distribution of Polystyrene Latex (PSL) particles. We investigate in particular some unique, nominally 200 nm and 100 nm PSL particles, known by other methods to be unusually monodisperse. The suspensions are electrosprayed after adding ammonium acetate to a concentration of 10 mmol/L. Mobility analysis is carried out after charge reduction with a second electrospray of the opposite polarity. The 200 nm particles typically produce two narrow peaks, with relative intensities that depend on electrospray parameters. Under the most favorable spraying conditions achieved, the more mobile peak becomes dominant, is narrowest, and has a repeatable mobility; we attribute it provisionally to the unpolluted suspended particles. The less mobile peaks are substantially more variable but never entirely removed. We initially attributed them to attachment of involatile solutes in the suspension to the atomized PSL particles; however, this hypothesis is put in doubt by later observations. The measured relative width of the mobility distribution for the most mobile peak is FWHMZ = (1.65 ± 0.08) %. This translates into a width for the diameter distribution FWHMd = (1.09 ± 0.05) % or standard deviation σd = (0.46 ± 0.02) %, by far the most monodisperse polymeric submicron particle suspension so far reported. Concentrations of ammonium acetate of 100 mmol/L and 40 mmol/L were also tested in order to reduce residue attachment by forming smaller initial electrosprayed drops. These higher salinities, however, led to capillary clogging, apparently because the narrow electrospraying jet precludes the passage of the PSL particles. The 100 nm particles gave a wider variation in mean diameter and size distribution depending on electrospraying parameters, perhaps because of the suspension's considerably larger content of involatile solutes. Unlike the 200 nm particles, no spraying condition was found under which a precisely repeatable size distribution could be achieved for the 100 nm particles. However, the data do point to an estimated upper bound on the size distribution width of FWHMd ≈ 3 % (σd ≈ 1.3 %), which is uniquely narrow for particles of this size.

Separation of disaccharide epimers, anomers and connectivity isomers by high resolution differential ion mobility mass spectrometry

Glycans are ubiquitous, structurally diverse molecules that have specific and general roles involving metabolism, structure, and cell-to-cell signaling. Functional specificity depends strongly on the complexity of structures that polysaccharides can adopt based on their subunit composition, length, extent of branching, glycosidic bond connectivity and anomeric configuration. However, a rapid and comprehensive characterization of glycan isomers can be challenging owing to limitations associated with their separation. Here, ten composition, anomeric and connectivity disaccharide isomers were separated and detected using high-resolution differential ion mobility-mass spectrometry (DMS-MS, also known as FAIMS). Focus was primarily directed to compositional isomers corresponding to epimers that differ by the axial or equatorial position of a single hydroxyl group. DMS resolving power was enhanced 14-fold primarily by increasing the fraction of helium in the ion carrier gas and lowering the flow rate. At relatively high disaccharide concentrations, DMS-MS of each disaccharide resulted in complex and unique multi-peak spectra with up to ten fully and partially resolved peaks for β-1,4-mannobiose (Man-1,4β-Man), which can be attributed to the DMS separation and subsequent dissociation of ionic non-covalently bound oligomers into monomer ions. Each DMS spectrum has at least one differentiating peak that is not in the other spectra, indicating that DMS can be used to fully or partially resolve composition, configuration and connectivity isomers. At relatively low disaccharide concentrations, mixtures of disaccharide epimers can also be readily separated by DMS. The integration of high-resolution, ambient pressure DMS with complementary reduced-pressure ion mobility and MS-based glycomics and glycoproteomics workflows may be useful for improving the characterization of glycans and glycosylated biomolecules.

An atmospheric pressure ion funnel with a slit entrance for enhancing signal and resolution in high resolution differential ion mobility mass spectrometry.

Differential ion mobility (DMS) is a versatile ion separation method that is often integrated with mass spectrometry (MS). In DMS, extremely high electric fields are used such that ion mobility depends non-linearly on electric field and thus, ion separations can be more orthogonal to MS than lower field ion mobility-based methods. DMS can have sufficiently high resolution to be used for enantiomer analysis of small molecules and to separate protein ions with peak widths comparable to those obtained for peptides. However, the performance of high resolution DMS-MS can be limited owing to the substantial loss of ions (>10-fold) that can occur upon their transfer from atmospheric pressure (where DMS separation typically occurs) to vacuum through a narrow conductance limited inlet (e.g. capillary) to the MS. Here, results from simulated ion trajectory simulations suggest that in high resolution DMS most ions can be lost by 'crashing' onto the narrow capillary inlet after exiting the DMS separation channel. To enhance DMS sensitivity and resolving power, an integrated DMS-MS interface concept is reported that consists of a slit electrode and a 12-electrode atmospheric pressure ion funnel (APIF). By using an APIF with slit entrance, the simulated ion transmission efficiencies increase by up to 257% for singly charged ions ([DMMP + H]+, [tryptophan + H]+, and [(2-dodecanone)2 + H]+) and by 209% for [ubiquitin + 12H]12+, without compromising resolving power. The use of APIF improves the ion focussing from the DMS exit to the MS capillary to improve sensitivity, and the slit ensures that ion dispersion in the analytically relevant direction perpendicular to the DMS electrodes is restricted to enhance resolution. By narrowing the slit of the DMS-Slit-APIF interface, the DMS resolving power can be increased further by at least 20%. Overall, these results indicate that the integrated DMS-Slit-APIF interface is promising for improving the sensitivity and resolution for many different types of DMS-MS experiments.

Disentangling Lipid Isomers by High-Resolution Differential Ion Mobility Spectrometry/Ozone-Induced Dissociation of Metalated Species.

The preponderance and functional importance of isomeric biomolecules have become topical in biochemistry. Therefore, one must distinguish and identify all such forms across compound classes, over a wide dynamic range as minor species often have critical activities. With all the power of modern mass spectrometry for compositional assignments by accurate mass, the identical precursor and often fragment ion masses render this task a steep challenge. This is recognized in proteomics and epigenetics, where proteoforms are disentangled and characterized employing novel separations and non-ergodic dissociation mechanisms. This issue is equally pertinent to lipidomics, where the lack of isomeric depth has thwarted the deciphering of functional networks. Here we introduce a new platform, where the isomeric lipids separated by high-resolution differential ion mobility spectrometry (FAIMS) are identified using ozone-induced dissociation (OzID). Cationization by metals (here K+, Ag+, and especially Cu+) broadly improves the FAIMS resolution of isomers with alternative C═C double bond (DB) positions or stereochemistry, presumably via metal attaching to the DB and reshaping the ion around it. However, the OzID yield diminishes for Ag+ and vanishes for Cu+ adducts. Argentination still strikes the best compromise between efficient separation and diagnostic fragmentation for optimal FAIMS/OzID performance.

Measurement of sub-2 nm stable clusters during silane pyrolysis in a furnace aerosol reactor.

The initial stages of particle formation are important in several industrial and environmental systems; however, the phenomenon is not completely understood due to the inability to measure cluster size distributions. A high resolution differential mobility analyzer with an electrometer was used to map out the early stages of Si particle formation from pyrolysis of SiH4 in a furnace aerosol reactor. We detected for the first time subnanometer stable clusters from silane pyrolysis, and the diameter was measured to be about 0.7 nm. This diameter is within the range of probable sizes that the reported families of critical silane clusters could have based on their actual molecular structure. The size distributions of negative clusters are also mapped out. In addition, gas chromatography mass spectrometry, and transmission electron microscopy characterizations of the clusters and primary particles are used to assess their mechanistic roles in aerosol dynamics of the initial stages of particle formation.

Analysis of ultrafine metal oxide particles in aerosols using mobility-resolved time-of-flight mass spectrometry

In this work we present a powerful method to gain insight into the formation of inorganic nanoparticles from the gas phase by combining a high-resolution differential mobility analyzer with a time-of-flight mass spectrometer. Exemplary particles are generated by glowing-wire evaporation from tungsten, tantalum, rhenium, niobium and molybdenum and transferred via a new custom-made interface to the mobility classifier that is directly attached to the mass spectrometer. With only minor fragmentation occurring, clusters that represent early stages of particle formation from the gas phase can be detected. Regardless of the inert experimental conditions, only oxides were found for all tested precursor metals. Comparing the recorded mass-mobility data with theoretical predictions from a modified Mason-Schamp equation shows that produced materials possess densities smaller than those of the respective bulk materials. The maximum observed particle sizes were about 2 nm.

High-Resolution Differential Ion Mobility Separations/Orbitrap Mass Spectrometry without Buffer Gas Limitations.

Strong orthogonality between differential ion mobility spectrometry (FAIMS) and mass spectrometry (MS) makes their hybrid a powerful approach to separate isomers and isobars. Harnessing that power depends on high resolution in both dimensions. The ultimate mass resolution and accuracy are delivered by Fourier Transform MS increasingly realized in Orbitrap MS, whereas FAIMS resolution is generally maximized by buffers rich in He or H2 that elevate ion mobility and lead to prominent non-Blanc effects. However, turbomolecular pumps have lower efficiency for light gas molecules and their flow from the FAIMS stage complicates maintaining the ultrahigh vacuum (UHV) needed for Orbitrap operation. Here we address this challenge via two hardware modifications: (i) a differential pumping step between FAIMS and MS stages and (ii) reconfiguration of vacuum lines to isolate pumping of the high vacuum (HV) region. Either greatly ameliorates the pressure increases upon He or H2 aspiration. This development enables free optimization of FAIMS carrier gas without concerns about MS performance, maximizing the utility and flexibility of FAIMS/MS platforms.

Rapid separation of isomeric perfluoroalkyl substances by high-resolution differential ion mobility mass spectrometry

The analysis of persistent organic pollutants, such as perfluoroalkyl substances (PFAS) including perfluorooctanoic acid (C7F15COOH, PFOA) and perfluorooctane sulfonate (C8F17SO3−, PFOS) by hyphenated chromatography-mass spectrometry methods is crucial in ensuring water quality. One of the challenges in PFOA and PFOS analysis is the separation of linear and branched isomers that have the same mass-to-charge ratio but can have different toxicological properties. Current methods that are used for separating isomeric PFAS require relatively long analysis times (min to h) that can limit throughput. An emerging technique for the direct analysis of isomeric compounds is differential mobility spectrometry (DMS), which can rapidly separate gas phase ions prior to detection by mass spectrometry (MS). However, an ion mobility-based method for the analysis of PFAS has not been reported in the literature. Herein, high-resolution DMS-MS is used to separate and detect isomeric PFAS compounds for the first time in a separation process that occurs in milliseconds. The resolution of isomeric peaks increased by over 200% and 500% for PFOA and PFOS isomers, respectively, using a DMS carrier gas composed of 50:50% He:N2 by volume compared to 100% N2, which was crucial in the separation of PFAS isomers under these conditions. Linear, secondary-branched, and tertiary-branched isomers of PFOA and PFOS including those that differ by the position of a single perfluoromethyl group can be resolved by DMS-MS. The DMS compensation field required to transmit different isomers increases as the extent of branching increases. For isomeric PFASs with a single branching point, the compensation field for optimal transmission also increases as the perfluoromethyl group is positioned closer to the carboxylate and sulfonate groups under these conditions. These results indicate that high-resolution DMS-MS should be a useful approach for the rapid analysis of isomeric PFAS.

Exploratory analysis of a sooting premixed flame via on-line high resolution (APi–TOF) mass spectrometry

By taking advantage of recent advances in High-Resolution Atmospheric Pressure intake Time of Flight (APi–TOF) Mass Spectrometry (MS), the chemical analysis of naturally charged flame-generated soot nuclei and precursors is explored using a well-characterized dilution sampling approach. Measurements were performed for mass-to-charge ratio up to 2000 Thomson, bridging the gap between the gas phase and the particle phase. The flame products were sampled at several heights above the burner (HAB) in the soot inception zone of the flame, quickly diluted in nitrogen and directly transported to the APi–TOF inlet. The investigated sooting premixed flame has been the object of multiple studies over the years and the present results complement existing literature data. The analyses of flame products naturally carrying charge of either polarity revealed the chemical and polarity-dependent complexity of the nucleation and chemi-ionization processes. The measured high-resolution mass spectra include peaks attributed to (hydrocarbon) molecules/clusters containing oxygen and nitrogen atoms and suggest that collision charging of flame pyrolysis products likely involves protonation/deprotonation of neutral materials. Results clearly show the change of the overall composition of the charged flame products at different HABs. Patterns in the mass spectra under different conditions were investigated to discriminate between collision charging, chemical reaction and physical clustering (i.e., coagulation and condensation) growth mechanisms. A comparison of the results with those obtained with High-resolution Differential Mobility Analysis (HR-DMA) in a recent study allowed for a more quantitative determination of the ion number concentrations.

Metal-ion free chiral analysis of amino acids as small as proline using high-definition differential ion mobility mass spectrometry

The chiral analysis of enantiomers is important because bioactivity can depend strongly on stereochemistry as ligand-protein binding motifs are typically chiral. Ion mobility mass spectrometry-based methods are emerging for the rapid and sensitive chiral analysis of molecules. However, such methods are typically limited by the use of metal-bound trimers, which can be challenging to form owing to ion suppression and the need for extensive pre-screening experiments to identify suitable metal ions. Moreover, the chiral separation of very small molecules, such as cysteine and proline, using ion mobility has remained challenging. Here, using electrospray ionisation high-resolution differential ion mobility mass spectrometry (ESI-DMS-MS), we demonstrate that the enantiomers of benchmark amino acids as small as proline can be rapidly distinguished without the use of metal ions for the first time. ESI-DMS-MS of proton-bound diastereomeric dimer complexes, containing enantiomers of amino acids and a ‘chiral selector’ (N-tert-butoxycarbonyl-O-benzyl-l-serine; BBS) corresponding to [L/D-X(BBS)+H]+ (X = cysteine and proline) resulted in the separation of L and D-enantiomers. By use of DMS-MS and standard solutions of chiral mixtures, these data indicate that the enantiomeric excess of proline can be accurately quantified by differential ion mobility mass spectrometry. Overall, these results provide further evidence that DMS-MS can be used for the rapid and accurate ‘metal-ion free’ chiral analysis of many other biologically important molecules.

Broad Separation of Isomeric Lipids by High-Resolution Differential Ion Mobility Spectrometry with Tandem Mass Spectrometry.

Maturation of metabolomics has brought a deeper appreciation for the importance of isomeric identity of lipids to their biological role, mirroring that for proteoforms in proteomics. However, full characterization of the lipid isomerism has been thwarted by paucity of rapid and effective analytical tools. A novel approach is ion mobility spectrometry (IMS) and particularly differential or field asymmetric waveform IMS (FAIMS) at high electric fields, which is more orthogonal to mass spectrometry. Here we broadly explore the power of FAIMS to separate lipid isomers, and find a ~75% success rate across the four major types of glycero- and phospho- lipids (sn, chain length, double bond position, and cis/trans). The resolved isomers were identified using standards, and (for the first two types) tandem mass spectrometry. These results demonstrate the general merit of incorporating high-resolution FAIMS into lipidomic analyses. Graphical Abstract ᅟ.

Probing gas-to-particle transition in a moderately sooting atmospheric pressure ethylene/air laminar premixed flame. Part II: Molecular clusters and nascent soot particle size distributions

A comprehensive investigation was conducted on a sooting ethylene laminar premixed flame. The present contribution focuses on measurements of soot nuclei by: a) thermophoretic sampling on fine wires followed by microscopy analysis and b) high-dilution probing followed by High-Resolution Differential Mobility Analysis (HR-DMA). The results complement Part I that was focused on the characterization of the gas composition and of soot properties by optical measurements, and highlighted significant artifacts induced by the heat loss associated with the presence of a stagnation plate. Significant novelties in the diagnostics include: the ability to perform spatially resolved thermophoretic measurements on fine wires straddling a critical temperature range that rules out artifacts brought about by either condensation of nonvolatiles or surface kinetics, as demonstrated by the invariance of particle size with the wire residence time in the flame; and the high-resolution capability of the DMA revealing the presence of small (D<3nm) nanoparticles both at lower heights in the flame, where younger soot is present, and further downstream at the same location where more mature soot is present. At the location at which both thermophoretically collected particles and HR-DMA data are available, the size distribution functions are in good agreement, except for the smallest particles that are quantifiable only by HR-DMA. Comparison of soot volume fraction and particle sizes from multiple techniques reveals good self-consistency, especially in the region where somewhat more mature soot particles are present (HAB≥12.5mm). A simplified three-dimensional model of the HR-DMA sampling procedure using a finite element solver package reveals the perturbation introduced by the “cold” probe to the temperature–time history experienced by the sampled aerosol and provides the input parameters for a one-dimensional streamtube model of the perturbed flame that included detailed chemistry. Even though additional work is needed to make the HR-DMA determination of the small particle peak fully quantitative, the developed database provides a useful benchmark for experimental validation of soot nucleation models, casting a shadow on established measurements that rely on commercial instrumentation with lower resolution.

The high charge fraction of flame-generated particles in the size range below 3 nm measured by enhanced particle detectors

Charging in flames significantly affects the properties of the resultant particles produced because of its influence in almost all stages of particle formation. The charging characteristics of flame-generated sub-3nm particles were investigated with three enhanced particle detectors including a high resolution differential mobility analyzer (DMA) coupled with an electrometer, a particle size magnifier coupled with a butanol-based condensation particle counter (PSM-bCPC), and an atmospheric pressure interface time-of-flight mass spectrometer (APi-TOF). Up to 95% of the flame-generated sub-3nm particles were charged at a sampling height of 5mm above the burner, indicating the existence of a strong ionization process in the investigated flame. This high fraction of charged particles contradicts the classical charging theories, which predict <1% charge fraction for particles below 3nm. Positively and negatively charged sub-3nm particles generated from a blank flame were dominated by organic ions and nitrate ions respectively. The flame-generated ions play an important role during titanium dioxide (TiO2) nanoparticle synthesis, as shown by the attachment of nitrate species on Ti-containing particles observed by the APi-TOF. The effects of the sampling height and precursor feed rate were also investigated.

Localization of Post-Translational Modifications in Peptide Mixtures via High-Resolution Differential Ion Mobility Separations Followed by Electron Transfer Dissociation.

Precise localization of post-translational modifications (PTMs) on proteins and peptides is an outstanding challenge in proteomics. While electron transfer dissociation (ETD) has dramatically advanced PTM analyses, mixtures of localization variants that commonly coexist in cells often require prior separation. Although differential or field asymmetric waveform ion mobility spectrometry (FAIMS) achieves broad variant resolution, the need for standards to identify the features has limited the utility of approach. Here we demonstrate full a priori characterization of variant mixtures by high-resolution FAIMS coupled to ETD and the procedures to systematically extract the FAIMS spectra for all variants from such data. Graphical Abstract ᅟ.

Observation of incipient particle formation during flame synthesis by tandem differential mobility analysis-mass spectrometry (DMA-MS)

While flame aerosol reactor (FLAR) synthesis of nanoparticles is widely used to produce a range of nanomaterials, incipient particle formation by nucleation and vapor condensation is not well understood. This gap in our knowledge of incipient particle formation is caused by limitations in instruments, where, during measurements, the high diffusivity of sub 3 nm particles significantly affects resolution and transport loss. This work used a high resolution differential mobility analyzer (DMA) and an atmospheric pressure interface-mass spectrometer (APi-TOF) to observe incipient particle formation during flame synthesis. By tandemly applying these two instruments, differential mobility analysis-mass spectrometry (DMA-MS) measured the size and mass of the incipient particles simultaneously, and the effective density of the sub 3 nm particles was estimated. The APi-TOF further provided the chemical compositions of the detected particles based on highly accurate masses and isotope distributions. This study investigated the incipient particle formation in flames with and without the addition of synthesis precursors. Results from FLAR using two types of precursors including tetraethyl orthosilicate (TEOS) and titanium isopropoxide (TTIP) are presented. The effect of the precursor feed rates on incipient particle growth was also investigated.

Challenges of measuring nascent soot in flames as evidenced by high-resolution differential mobility analysis

ABSTRACTNascent soot particles with mobility diameters ≤10 nm were measured in an ethylene/air premixed flame to shed light on the challenges and potential artifacts affecting studies on soot inception by differential mobility analysis (DMA) techniques. The size distribution functions (SDFs) of particles with charge acquired either naturally or diffusively upon ion seeding were measured at several positions in the flame using rapid-dilution probing and a high-resolution DMA for different values of the ratio of dilution ratio to residence time (DR/Δt). The SDFs are roughly bimodal with a sub-3 nm mode and a larger one that appears either downstream in the flame or for low DR/Δts. Soot nuclei smaller than 3 nm preferentially acquire positive charge, which brings into question the assumption of steady-state charging probability of flame sampled soot nuclei in the bipolar diffusion neutralizer. The approximately polarity-symmetric lognormal SDF of larger particles is attributed to nuclei coagulation. Naturall...