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Investigation of passive atmospheric sounding using millimeter and submillimeter wavelength channels

Presented in this study are the results of controlled partially polarimetric measurements of thermal emission at 91.65 GHz from a striated water surface as corroborated by a geometrical optics radiative model. The measurements were obtained outdoors using a precision polarimetric radiometer which directly measured the first three modified Stokes' parameters. Significant variations in these parameters as a function of azimuthal water wave angle were found, with peak-to-peak variations in T(sub u) of up to approximately 10 K. The measurements are well corroborated by the GO model over a range of observations angles from near nadir up to approximately 65 degrees from nadir. The model incorporates both multiple scattering and a realistic downwelling background brightness field.

Gasiewski, Albin J.↗

Analysis of Small-Scale Atmospheric Gravity Waves Using UARS MLS Radiance Measurements

Gravity waves play an important role in determining atmospheric circulation and small-scale mixing. Upper Atmosphere Research Satellite (UARS) Microwave Limb Sounder (MLS) 63-GHz radiances can be used to calculate small-scale wave variances at 30-80 km altitudes. The major results from this new data set are summarized in the following: (1) MLS radiance fluctuations are contributed mostly by gravity waves of small (about 100 km) horizontal and large (>10 km) vertical scales. (2) MLS observations show that variance enhancements are strongly correlated with the stratospheric polar vortices, tropospheric deep convection zones, and surface topography. (3) As expected for gravity wave propagation, the normalized wave variances grow exponentially with height in the stratosphere but saturate in the mesosphere. (4) The long-term variations of the wave variance are dominated by an annual cycle in the stratosphere and a semiannual cycle in the mesosphere. (5) Separate analyses of the ascending and descending measurements show that the variances are sensitive to wave propagation directions. The subtropical variances, which are associated with deep convection, are likely caused by the gravity waves that propagate upward and eastward in the westward background wind. Additional information contained in the original.

Wu, Dong L.↗

(abstract) A Polarimetric Model for Effects of Brine Infiltrated Snow Cover and Frost Flowers on Sea Ice Backscatter

A polarimetric scattering model is developed to study effects of snow cover and frost flowers with brine infiltration on thin sea ice. Leads containing thin sea ice in the Artic icepack are important to heat exchange with the atmosphere and salt flux into the upper ocean. Surface characteristics of thin sea ice in leads are dominated by the formation of frost flowers with high salinity. In many cases, the thin sea ice layer is covered by snow, which wicks up brine from sea ice due to capillary force. Snow and frost flowers have a significant impact on polarimetric signatures of thin ice, which needs to be studied for accessing the retrieval of geophysical parameters such as ice thickness. Frost flowers or snow layer is modeled with a heterogeneous mixture consisting of randomly oriented ellipsoids and brine infiltration in an air background. Ice crystals are characterized with three different axial lengths to depict the nonspherical shape. Under the covering multispecies medium, the columinar sea-ice layer is an inhomogeneous anisotropic medium composed of ellipsoidal brine inclusions preferentially oriented in the vertical direction in an ice background. The underlying medium is homogeneous sea water. This configuration is described with layered inhomogeneous media containing multiple species of scatterers. The species are allowed to have different size, shape, and permittivity. The strong permittivity fluctuation theory is extended to account for the multispecies in the derivation of effective permittivities with distributions of scatterer orientations characterized by Eulerian rotation angles. Polarimetric backscattering coefficients are obtained consistently with the same physical description used in the effective permittivity calculation. The mulitspecies model allows the inclusion of high-permittivity species to study effects of brine infiltrated snow cover and frost flowers on thin ice. The results suggest that the frost cover with a rough interface significantly increases the backscatter from thin saline ice and the polarimetric signature becomes closer to the isotropic characteristics. The snow cover also modifies polarimetric signatures of thin sea ice depending on the snow mixture and the interface condition.

thin sea ice radar backscatter frost flowers snow ↗

Sample mounts for microcrystal crystallography

Sample mounts (10) for mounting microcrystals of biological macromolecules for X-ray crystallography are prepared by using patterned thin polyimide films (12) that have curvature imparted thereto, for example, by being attached to a curved outer surface of a small metal rod (16). The patterned film (12) preferably includes a tip end (24) for holding a crystal. Preferably, a small sample aperture is disposed in the film for reception of the crystal. A second, larger aperture can also be provided that is connected to the sample aperture by a drainage channel, allowing removal of excess liquid and easier manipulation in viscous solutions. The curvature imparted to the film (12) increases the film's rigidity and allows a convenient scoop-like action for retrieving crystals. The polyimide contributes minimally to background and absorption, and can be treated to obtain desired hydrophobicity or hydrophilicity.

Thorne, Robert E.↗

Wasserstein Normalized Autoencoder for Anomaly Detection in ProtoDUNE Vertical-Drift Detector

ProtoDUNE Vertical Drift needs a selective triggering algorithm. The detector sits on Earth's surface, so cosmic activity dominates its data. Our goal in this paper is to trigger on neutrino events more robustly than the current deployed Analog-to-Digital Converter Simple Window (ADCSW) model and, eventually, search for signals of Beyond Standard Model (BSM) physics at DUNE as our ultimate North Star objective. As a step towards this goal, we evaluate a Wasserstein Normalized Autoencoder (WNAE) on simulated collection-plane only windows of shape $1\times10\times10$ where Neutrinos act as our BSM-proxy and Cosmic-ray Muons serve as our learned background. The network parameters are fitted using only cosmic-ray muon events as background in order to maintain an unsupervised pipeline. Training uses finite-step Langevin $x^-$ samples, positive-sample reconstruction energy, and an empirical sliced $2$-Wasserstein objective to learn a normalized Boltzmann energy model. We then calibrate on a nominal $5\,\mathrm{Hz}$ operating threshold calculated from cosmic validation data. Both WNAE and ADCSW accept 311 of 194,083 held-out cosmic background events at this $5\,\mathrm{Hz}$ threshold. We found that WNAE accepts 9,677 of 34,634 neutrino-proxy events $(27.9\pm0.24)\%$, compared with 10,076 $(29.1\pm0.24)\%$ for ADCSW, an observed WNAE-minus-ADCSW difference of $-1.15\%$. At another nominal $2\,\mathrm{Hz}$ target threshold, the corresponding efficiencies are $(20.5\pm0.22)\%$ and $(22.6\pm0.22)\%$, respectively. Of the WNAE-selected neutrino proxies at $5\,\mathrm{Hz}$, $(20.8\pm0.4)\%$ of the classified neutrino-proxy events are unique to WNAE, where the uncertainty is an absolute binomial standard error of $0.4\%$.

Zheng, Jake [Chicago U.] (ORCID:0009000218901379)↗

Ensemble Spread Behavior in Coupled Climate Models: Insights From the Energy Exascale Earth System Model Version 1 Large Ensemble

AbstractAssessing uncertainty in future climate projections requires understanding both internal climate variability and external forcing. For this reason, single‐model initial condition large ensembles (SMILEs) run with Earth System Models (ESMs) have recently become popular. Here we present a new 20‐member SMILE with the Energy Exascale Earth System Model version 1 (E3SMv1‐LE), which uses a “macro” initialization strategy choosing coupled atmosphere/ocean states based on inter‐basin contrasts in ocean heat content (OHC). The E3SMv1‐LE simulates tropical climate variability well, albeit with a muted warming trend over the twentieth century due to overly strong aerosol forcing. The E3SMv1‐LE's initial climate spread is comparable to other (larger) SMILEs, suggesting that maximizing inter‐basin ocean heat contrasts may be an efficient method of generating ensemble spread. We also compare different ensemble spread across multiple SMILEs, using surface air temperature and OHC. The Community Earth system Model version 1, the only ensemble which utilizes a “micro” initialization approach perturbing only atmospheric initial conditions, yields lower spread in the first ∼30 years. The E3SMv1‐LE exhibits a relatively large spread, with some evidence for anthropogenic forcing influencing spread in the late twentieth century. However, systematic effects of differing “macro” initialization strategies are difficult to detect, possibly resulting from differing model physics or responses to external forcing. Notably, the method of standardizing results affects ensemble spread: control simulations for most models have either large background trends or multi‐centennial variability in OHC. This spurious disequlibrium behavior is a substantial roadblock to understanding both internal climate variability and its response to forcing.

Stevenson, Samantha↗

Meso-beta scale numerical simulation studies of terrain-induced jet streak mass/momentum perturbations

Work performed during the report period is summarized. The first numerical experiment which was performed on the North Carolina Supercomputer Center's CRAY-YMP machine during the second half of FY92 involved a 36 hour simulation of the CCOPE case study. This first coarse-mesh simulation employed the GMASS model with a 178 x 108 x 32 matrix of grid points spaced approximately 24 km apart. The initial data was comprised of the global 2.5 x 2.5 degree analyses as well as all available North American rawinsonde data valid at 0000 UTC 11 July 1981. Highly-smoothed LFM-derived terrain data were utilized so as to determine the mesoscale response of the three-dimensional atmosphere to weak terrain forcing prior to including the observed highly complex terrain of the northern Rocky Mountain region. It was felt that the model should be run with a spectrum of terrain geometries, ranging from observed complex terrain to no terrain at all, to determine how crucial the terrain was in forcing the mesoscale phenomena. Both convection and stratiform (stable) precipitation were not allowed in this simulation so that their relative importance could be determined by inclusion in forth-coming simulations. A full suite of planetary boundary layer forcing was allowed in the simulation, including surface sensible and latent heat fluxes employing the Blakadar PBL formulation. The details of this simulation, which in many ways could be considered the control simulation, including the important synoptic-scale, meso-alpha scale, and meso-beta scale circulations is described. These results are compared to the observations diagnosed by Koch and his colleagues as well as hypotheses set forth in the project proposal for terrain-influences upon the jet stream and their role in the generation of mesoscale wave phenomenon. The fundamental goal of the analyses being the discrimination among background geostrophic adjustment, terrain influences, and shearing instability in the initiation and maintainance of mesoscale internal wave phenomena. Based upon these findings, FY93 plans are discussed. A review of linear theory and theoretical modeling of a geostrophic zonal wind anomaly is included.

Lin, Yuh-Lang↗

Detection of CO emission from massive molecular clouds in the inner disk of M31

Faint emission in the CO(1-0) and CO(2-1) spectral lines at mm wavelengths has been discovered from dark dust clouds in the inner disk of M31. These dust clouds are located in regions of the galaxy where there is little evidence of star-forming activity. The CO emission from these clouds is resolved, and comes from roughly the same area covered by the dust clouds. The line profile widths and cloud sizes imply cloud masses typical for giant molecular clouds (GMCs) in the Galaxy, but the observed surface brightnesses are only a few tenths of a K, and the typical CO(1-0) luminosities are a factor of about 10 smaller than those of Galactic GMCs of the same size and velocity line width. The CO(2-1)/CO(1-0) line ratios are very low, suggesting subthermal excitation, and precluding use of this ratio alone to determine the kinetic temperature. However, there are indications that the kinetic temperatures are significantly lower than the values of 10-30 K typical for Galactic GMCs, and may be close to the 2.7 K cosmic background.

Allen, Ronald J.↗

Spherule Beds 3.47-3.24 Billion Years Old in the Barberton Greenstone Belt, South Africa: A Record of Large Meteorite Impacts and Their Influence on Early Crustal and Biological Evolution

Four layers, S1-S4, containing sand-sized spherical particles formed as a result of large meteorite impacts, occur in 3.47-3.24 Ga rocks of the Barberton Greenstone Belt, South Africa. Ir levels in S3 and S4 locally equal or exceed chondritic values but in other sections are at or only slightly above background. Most spherules are inferred to have formed by condensation of impact-produced rock vapor clouds, although some may represent ballistically ejected liquid droplets. Extreme Ir abundances and heterogeneity may reflect element fractionation during spherule formation, hydraulic fractionation during deposition, and/or diagenetic and metasomatic processes. Deposition of S1, S2, and S3 was widely influenced by waves and/or currents interpreted to represent impact-generated tsunamis, and S1 and S2 show multiple graded layers indicating the passage of two or more wave trains. These tsunamis may have promoted mixing within a globally stratified ocean, enriching surface waters in nutrients for biological communities. S2 and S3 mark the transition from the 300-million-year-long Onverwacht stage of predominantly basaltic and komatiitic volcanism to the late orogenic stage of greenstone belt evolution, suggesting that regional and possibly global tectonic reorganization resulted from these large impacts. These beds provide the oldest known direct record of terrestrial impacts and an opportunity to explore their influence on early life, crust, ocean, and atmosphere. The apparent presence of impact clusters at 3.26-3.24 Ga and approx. 2.65-2.5 Ga suggests either spikes in impact rates during the Archean or that the entire Archean was characterized by terrestrial impact rates above those currently estimated from the lunar cratering record.

Lowe, Donald R.↗

Sample mounts for microcrystal crystallography

Sample mounts (10) for mounting microcrystals of biological macromolecules for X-ray crystallography are prepared by using patterned thin polyimide films (12) that have curvature imparted thereto, for example, by being attached to a curved outer surface of a small metal rod (16). The patterned film (12) preferably includes a tapered tip end (24) for holding a crystal. Preferably, a small sample aperture is disposed in the film for reception of the crystal. A second, larger aperture can also be provided that is connected to the sample aperture by a drainage channel, allowing removal of excess liquid and easier manipulation in viscous solutions. The curvature imparted to the film (12) increases the film's rigidity and allows a convenient scoop-like action for retrieving crystals. The polyimide contributes minimally to background and absorption, and can be treated to obtain desired hydrophobicity or hydrophilicity.

Thorne, Robert E.↗

Effective entropy of quantum fields coupled with gravity

Entanglement entropy, or von Neumann entropy, quantifies the amount of uncertainty of a quantum state. For quantum fields in curved space, entanglement entropy of the quantum field theory degrees of freedom is well-defined for a fixed background geometry. In this paper, we propose a generalization of the quantum field theory entanglement entropy by including dynamical gravity. The generalized quantity named effective entropy, and its Renyi entropy generalizations, are defined by analytic continuation of a replica calculation. The replicated theory is defined as a gravitational path integral with multiple copies of the original boundary conditions, with a co-dimension-2 brane at the boundary of region we are studying. We discuss different approaches to define the region in a gauge invariant way, and show that the effective entropy satisfies the quantum extremal surface formula. When the quantum fields carry a significant amount of entanglement, the quantum extremal surface can have a topology transition, after which an entanglement island region appears. Our result generalizes the Hubeny-Rangamani-Takayanagi formula of holographic entropy (with quantum corrections) to general geometries without asymptotic AdS boundary, and provides a more solid framework for addressing problems such as the Page curve of evaporating black holes in asymptotic flat spacetime. We apply the formula to two example systems, a closed two-dimensional universe and a four-dimensional maximally extended Schwarzchild black hole. We discuss the analog of the effective entropy in random tensor network models, which provides more concrete understanding of quantum information properties in general dynamical geometries. We show that, in absence of a large boundary like in AdS space case, it is essential to introduce ancilla that couples to the original system, in order for correctly characterizing quantum states and correlation functions in the random tensor network. Using the superdensity operator formalism, we study the system with ancilla and show how quantum information in the entanglement island can be reconstructed in a state-dependent and observer-dependent map. We study the closed universe (without spatial boundary) case and discuss how it is related to open universe.

79 ASTRONOMY AND ASTROPHYSICS↗

Hyper-Velocity Nanoparticle Plasma Jet as Fast Probe for Runaway Electrons in Tokamak Disruptions

The large energy focalized deposition of the runaway electron (RE) beam on the first wall can potentially yield to destructive effects on the reactor vessel during the explosive instability of a major disruption in a large tokamak (e.g. RE beam estimation is multi-MA of ~10 MeV energy in ITER.) Impurity injection is seen as an effective way to suppress REs after thermal quench (TQ). Injected material in solid state is advantageous, but it must be in small size grain form, with large surface-to-mass ratio to facilitate its rapid ablation within plasma. However, the results using shattered pellet injection (SPI), a technique aiming to satisfy these condition, indicated that the relatively unexplored RE evolution phases and RE beam-plasma interaction are far from being properly diagnosed and fully understood. We proposed the idea to probe REs with a pulsed power hyper-velocity large-mass nanoparticle plasma jet (NPPJ). NPPJ provides a high specific surface area and swift injection to core, hence fast increase of three basic parameters: 1) the electron density ne (bound and free through ionization, if the cold post-TQ plasma has a still high enough T e ), 2) the effective charge (Z eff ) which rises both the dynamical friction force and plasma resistivity, and, finally, 3) the value of the (Dreicer and critical) threshold E-field above which electrons are accelerated and become REs. But the fast transient phenomena of RE ”seed” creation, RE ”avalanche” multiplication, and RE beam formation and interaction with the background plasma all take place dominantly in the core plasma, where the induction E-field has its maximum. Thus, the RE probe success requires a rapid (1 - 2 ms) injection of sufficient mass able to penetrate through the strong toroidal magnetic field (2 - 5 T) over a long distance (~1 - 2 m) with a large assimilation fraction in the core plasma. As a RE probe, NPPJ is a unique combination of fast response with trigger-to-delivery time of few milliseconds, large mass-velocity (>75 mg at several km/s) and deep injection into post-TQ central plasma. The Objectives of this Category #1 project (concept development) were as follows: 1: We investigated the essential basic physics aspects of creating a plasma with ~10 15 to 10 16 cm -3 boron nitride (BN) NP density suited for a pulsed power plasma gun acceleration to several km/s. We concluded that, for RE NPPJ probe, BN (diameter Ø10 to 100 nm NPs) are in all respects usable. 2: We advanced solutions to get BN NPPJ by adapting FAR-TECH’s technology for C 60 NPPJ. 3: We developed self-consistent models for RE current density j RE (R; t) evolution in the TQ aftermath, with fast and simultaneous increase of n e (R; t) and Z eff (R; t) > 1 upon the specific injection of C 60 (i.e. C ions) and BN NP (i.e. B and N ions), respectively. We showed that prompt post-TQ injection and high velocity are of essence in RE probe diagnostics and suppression. 4: We used the in-house HEM-2D code to simulate the C$_{60}^{q+}$ NPPJ transverse penetration through tokamak-level B-field (BT ~2 to 5 T) and distance (~1 to 2m), with fragmentation of C$_{60}^{q+}$ and developed the algorithm for BN Q+ NPPJ ablative sublimation of BN Q+ NP by the collisions with post-TQ plasma electrons. HEM-2D results indicated that fragmenting C$_{60}^{1+}$ NPPJ penetrates up to ~60 cm in 100 us when injecting at 1.6 ms after TQ and gradually released C 2 fragments and C ions.

70 PLASMA PHYSICS AND FUSION TECHNOLOGY↗

Opportunities at the Sanford Underground Research Facility

The Sanford Underground Research Facility (SURF) has been operating for more than 15 years as an international facility dedicated to advancing compelling multidisciplinary underground scientific research in rare-process physics, as well as offering research opportunities in other disciplines. SURF laboratory facilities include a Surface Campus as well as campuses at the 4850-foot level (1490 m, 4300 m.w.e.) that host a range of significant physics experiments, including the LUX-ZEPLIN (LZ) dark matter experiment and the MAJORANA DEMONSTRATOR neutrinoless double-beta decay experiment. The CASPAR nuclear astrophysics accelerator completed the first phase of operation and is planning for the second phase beginning in 2024. SURF is also home to the Long-Baseline Neutrino Facility (LBNF) that will host the international Deep Underground Neutrino Experiment (DUNE). SURF offers world-class service, including an ultra-low background environment, low-background assay capabilities, and electroformed copper is produced at the facility. SURF is preparing to increase underground laboratory space. Plans are advancing for construction of new large caverns (nominally 100m L x 20m W x 24m H) on the 4850L (1485 m, 4100 mwe) on the timeframe of next-generation experiments (~2030). SURF plans to leverage existing advisory and community committees as well as engage the underground science community to inform plans for future laboratory space.

72 PHYSICS OF ELEMENTARY PARTICLES AND FIELDS↗

Project 1640 Observations of Brown Dwarf GJ 758 B: Near-infrared Spectrum and Atmospheric Modeling

The nearby Sun-like star GJ 758 hosts a cold substellar companion, GJ 758 B, at a projected separation of ≲30 au, previously detected in high-contrast multi-band photometric observations. In order to better constrain the companion’s physical characteristics, we acquired the first low-resolution (R ∼ 50) near-infrared spectrum of it using the high-contrast hyperspectral imaging instrument Project 1640 on Palomar Observatory’s 5 m Hale telescope. We obtained simultaneous images in 32 wavelength channels covering the Y, J, and H bands (∼952–1770 nm), and used data processing techniques based on principal component analysis to efficiently subtract chromatic background speckle-noise. GJ 758 B was detected in four epochs during 2013 and 2014. Basic astrometric measurements confirm its apparent northwest trajectory relative to the primary star, with no clear signs of orbital curvature. Spectra of SpeX/IRTF observed T dwarfs were compared to the combined spectrum of GJ 758 B, with χ {sup 2} minimization suggesting a best fit for spectral type T7.0 ± 1.0, but with a shallow minimum over T5–T8. Fitting of synthetic spectra from the BT-Settl13 model atmospheres gives an effective temperature T {sub eff} = 741 ± 25 K and surface gravity logg=4.3±0.5 dex (cgs). Our derived best-fit spectral type and effective temperature from modeling of the low-resolution spectrum suggest a slightly earlier and hotter companion than previous findings from photometric data, but do not rule out current results, and confirm GJ 758 B as one of the coolest sub-stellar companions to a Sun-like star to date.

79 ASTRONOMY AND ASTROPHYSICS↗

Electromagnetic evidence concerning the lunar interior and its evolution

Reviews of the evidence for magnetization of the moon found from discovery of remanence in lunar samples, direct measurements of fields on the surface of the moon, and direct and indirect determination of fields from lunar orbit. It is shown that the evidence implies that the fields are not only local but that regional properties are found though there is still no direct evidence for a global dipole moment. Limits on the detectibility of a global dipole are given and it is shown that the strength of magnetization for reasonable thermal gradients places possible dipole moments just below the threshold of detectibility of current experiments. The hypothesis of plate magnetics is reviewed. Current ideas regarding the source of the background magnetic field presumed responsible for the magnetization are critically considered. These are the dynamo hypothesis and primordial magnetization. Consequences of both are discussed and finally the constraints placed upon the thermal evolution of the moon are considered.

Sonett, C. P.↗

Modelling of the MEA float zone using accelerometer data

During a floating zone experiment involving the growth of indium on a recent orbiter mission, (STS 32) oscillation of the zone shapes were observed to occur in response to the background acceleration. An understanding of the nature of the response of the zone shape to forced (g-jitter) oscillations and predictions of its impact on future experiments is of great interest not only to the PI's but to other commercial and academic investigators who plan to fly similar experiments in the orbiter and on space station. Motivated by this, a 15 month study was undertaken to analyze the nature of the g-sensitivity of the STS 32 floating zone crystal growth experiment. Numerical models were used to describe the time-dependent free surface motion of the zone as it responds to the spacecraft residual acceleration. Relevant experimental data concerning the acceleration environment was obtained from the Honeywell in Space Accelerometer (HISA) investigators through MSFC's ACAP program and processed and analyzed. For the indium floating zone experiment, a series of calculations were made using time-dependent axial accelerations g(t). The form of g(t) included simple sinusoidal disturbances as well as actual data (subject to appropriate filtering) measured on the STS 32 mission. Focus was on the calculation of the response of the free surface of the zone as well as the internal flows and internal heat transfer. The influence of solidification on the response of the zone shape was also examined but found to be negligible.

Alexander, J. Iwan D.↗

Investigation of the effect of noise parameters of a 3D lidar on the error in estimating relief signatures of distant objects from 2D field intensity distributions of reflected radiation

We consider the influence of noise parameters of a 3D lidar and external background radiation on the error in estimating the relief signatures of distant objects from 2D field intensity distributions of the reflected radiation. A physical and mathematical model is developed to analyse the effect of noise. The dependence of the error in estimating the relief signatures at various signal-to-noise ratios is investigated taking into account the parameters of real multi-element photodetectors (MEPDs). The obtained results indicate the possibility of evaluating relief signatures of distant objects with high accuracy based on standard MEPDs simultaneously for a large number of points on the object’s surface with a signal-to-noise ratio of more than 4. (paper)

46 INSTRUMENTATION RELATED TO NUCLEAR SCIENCE AND ↗

Calibration of 2-D soil zymography for correct analysis of enzyme distribution

Soil zymography is a new technique developed to visualize two-dimensional distributions of enzyme activities. The method is composed of incubating a membrane saturated with an enzyme-specific fluorogenic substrate on a surface of the soil sample, followed by recording the membrane image generated by a fluorescent product (e.g. MUF: methylumbelliferone) in ultraviolet light. Despite its relative ease of use, performing zymography involves multiple user-made decisions that might affect the accuracy of enzyme activity estimates. Thus, unification of the zymography methodology is required for correct estimations and comparisons of various studies. We investigated the following methodological aspects of the implementation of zymography: (a) camera settings and image processing, (b) effects of evaporation and (c) calibration procedures. Camera settings (shutter speeds or exposure time) affected the intensity of background fluorescence and signal-to-noise ratios (SNR). However, because their combined effects varied depending on MUF concentrations, light and camera setting need to be optimized for the expected range of MUF concentrations prior to zymography. Evaporation of MUF solution from the membrane had no effect on fluorescence. Relations between MUF concentration and intensity of fluorescence during calibrations demonstrated a saturated pattern and were strongly affected by image noise outside the optimal range (e.g. 8–14 μm MUF pixel -1 ). We developed a new calibration approach that is based on a piecewise linear regression. The new approach accounted for specific ranges of MUF concentration and uses nonuniformly saturated membranes, reflecting the real distribution of enzyme activities in soil. The new calibration algorithm eliminated biases of the standard calibration and resulted in greater accuracy in predicting MUF concentrations.

59 BASIC BIOLOGICAL SCIENCES↗