Fluorescence Line Research Articles

Synoptic-to-intraseasonal atmospheric modulation of phytoplankton biomass in the inner sea of Chiloé, Northwest Patagonia (42.5°-43.5°S, 72.5°-74°W), Chile

The Inner Sea of Chiloé (ISC) in northwestern Patagonia has experienced large harmful algal blooms in the past decade, impacting human health and affecting the large aquaculture industry of the region. Thus, the investigation of factors favouring regional phytoplankton growth are of particular interest. Analysing the synoptic-to-intraseasonal variability, we explore changes in phytoplankton biomass in southern ISC (S-ISC, 42.5°-43.5°S, 72.5°-74°W) and their concurrent mesoscale and large-scale meteorological and oceanographic conditions. We use high-resolution satellite normalized fluorescence line height (nFLH) and chlorophyll-a (CHL-A) from the MODIS-Aqua sensor as proxies for phytoplankton biomass, besides oceanic and atmospheric variables derived from various remote-sensing sources and atmospheric fields from the ERA5 reanalysis. Specifically, we focus on high phytoplankton biomass events HBEs, which are defined as those cases when intraseasonal nFLH anomaly (nFLH’) exceeds the 95th percentile threshold. Each event was characterised by its first date of occurrence (called day 0). We detected 16 HBE between 2003 and 2019 in S-ISC. HBEs tend to occur under the influence of a mid-latitude migratory anticyclone that induce persistent cloudless conditions preceding day 0, leading to enhanced photosynthetically active radiation (PAR) starting around day -8, and positive sea surface temperature (SST) anomalies between days -4 and +4. We hypothesise that HBEs are mainly modulated by i) mixing and advection that could contribute to a greater availability of nutrients in the upper sea layers before the onset of the anticyclonic anomalies; and ii) increased thermal stratification related to positive PAR and SST anomalies that would promote phytoplankton growth during the anticyclonic regime. Furthermore, we show that the Madden-Julian Oscillation modulates the frequency of nFLH’ and thus of HBEs, a result that suggests an enhanced predictability of these cases.

Construction of fluorescent transgenic zebrafish Tg (ttn.2: EGFP)

In order to develop a transgenic zebrafish line with green fluorescent protein (enhanced green fluorescent protein, EGFP) expressed specifically in muscle and heart, the recombinant expression vector constructed using the zebrafish ttn.2 gene promoter fragment and EGFP gene coding sequence and the capped mRNA of Tol2 transposase were co-injected into the zebrafish 1-cell stage embryos. The stable genetic Tg (ttn.2: EGFP) transgenic zebrafish line was successfully developed by fluorescence detection, followed by genetic hybridization screening and molecular identification. Fluorescence signals and whole-mount in situ hybridization showed that EGFP expression was located in muscle and heart, the specificity of which was consistent with the expression of ttn.2 mRNA. Inverse PCR showed that EGFP was integrated into chromosomes 4 and 11 of zebrafish in No. 33 transgenic line, while integrated into chromosome 1 in No. 34 transgenic line. The successful construction of this fluorescent transgenic zebrafish line, Tg (ttn.2: EGFP), laid a foundation for the research of muscle and heart development and related diseases. In addition, the transgenic zebrafish lines with strong green fluorescence can also be used as a new ornamental fish.

Nustar observation of the binary system 4U 0114 + 65

ABSTRACT The high-mass X-ray binary system 4U 0114 + 65 was observed by Nustar in October 2019 and by XMM-Newton in August 2015. Here we performed spectral and timing analysis of the Nustar observation and carry out timing analysis on the XMM-Newton data. We measured the spin period of the neutron star from both observations and found a spin-up rate $\dot{p}$ = 1.54 ± 0.38 × 10−6$\mathrm{s\, s}^{-1}$ . During the Nustar observation, two flares occurred, one shortly after the start of the observation and the other near the end separated by a long period of low/quiescent state. The large and sudden flares mostly resulted from accretion of corotating interaction region (CIR) material. A common spectral model to high mass X-ray binaries (HMXBs), powerlaw with high energy cutoff and absorption at low energy, gave a good fit to both flaring and quiescent states. A fluorescent iron line was not required in fitting any of the states. On the other hand, very tentative evidence of Cyclotron Resonant Scattering Feature (CRSF) at ∼17 keV was found during fitting using cyclabs model; however, fitting improvement was not significant enough to confirm its detection, plus a very narrow width (<1 keV) was obtained for the line and its first harmonic. Visual inspection of the spectra showed a deficiency of emission near the expected first and second harmonic. Another important feature visually noticed in the spectra is the presence of hard tail above 50 keV. This could be explained by the shocked material bounding the CIR.

Seeded stimulated X-ray emission at 5.9 keV.

X-ray free-electron lasers (XFELs) provide intense pulses that can generate stimulated X-ray emission, a phenomenon that has been observed and studied in materials ranging from neon to copper. Two schemes have been employed: amplified spontaneous emission (ASE) and seeded stimulated emission (SSE), where a second color XFEL pulse provides the seed. Both phenomena are currently explored for coherent X-ray laser sources and spectroscopy. Here, we report measurements of ASE and SSE of the 5.9 keV Mn Kα1 fluorescence line from a 3.9 molar NaMnO4 solution, pumped with 7 femtosecond FWHM XFEL pulses at 6.6 keV. We observed ASE at a pump pulse intensity of 1.7 × 1019 W/cm2, consistent with earlier findings. We observed SSE at dramatically reduced pump pulse intensities down to 1.1 × 1017 W/cm2. These intensities are well within the range of many existing XFEL instruments, which supports the experimental feasibility of SSE as a tool to generate coherent X-ray pulses, spectroscopic studies of transition metal complexes, and other applications.

On the Zeeman effect in magnetically arrested disks

Abstract The magnetically arrested disk (MAD) scenario has been argued as the key accretion phase to realize the formation of relativistic jets. However, due to the lack of magnetic field measurements of accreting systems, MAD has not been observationally confirmed yet. Here we propose that a strong magnetic field accompanied by MAD would induce the Zeeman splitting of relativistically broadened Fe Kα fluorescence lines in X-ray binaries and active galactic nuclei, where we consider a two-phase medium in the inner accretion disk, magnetically dominated hot corona, and cold reflector. Such a geometrical configuration is suggested from X-ray observations and recently confirmed by numerical simulations. Although turbulence in accretion flows would broaden the split lines, future X-ray high-energy resolution satellites, XRISM and Athena, would be capable of seeing the Zeeman effect on the Fe lines in X-ray binaries in the case with the MAD configuration. The signature of the Zeeman split lines would provide observational evidence for MAD.

Ultrasensitive Hybridization Chain Reaction-Assisted Multisite Exonuclease III Amplification Strategy Combined with a Direct Quantitative Fluorescence Lateral Flow Technique for Multiple Bacterial 16S rRNA Detection.

Accurate and in-time detection of bacteria conduces to preventing their rapid spread around the environment, while a nucleic acid test (NAT) is a powerful tool for early diagnosis of pathogens. Herein, we propose a hybridization chain reaction (HCR)-mediated multisite exonuclease III (Exo-III) amplification strategy (HCR/Exo-III amplifier) to achieve the one-pot and ultrasensitive isothermal amplification of bacterial 16S rRNA and a portable fluorescence detection device (PFD) to directly read signals in a lateral flow assay (LFA). In detail, the target-initiated HCR products present multiple binding sites for triggering the Exo-III amplifier that produces numerous target amplicons. Following that, the target amplicons travel up on the strip and bridge between the DNA-CdTe/CdS probes and the capture DNA to form a positive fluorescence line. After that, the strip is inserted into the PFD to accomplish the fluorescence signal reading. The constructed HCR/Exo-III amplifier-based PFD-LFA implemented the simultaneous and specific detection of three bacteria with a detection limit of a few tenths of fM for synthetic 16S rRNA fragments and dozens of CFU/mL for Staphylococcus aureus, Listeria monocytogenes, and Salmonella typhimurium in pure cultures. The sensing platform features isothermal amplification, convenient operation, and good economy, displaying great potential for on-site testing toward multiple nucleic acid analytes.

NGC 4388: a test case for relativistic disc reflection and Fe K fluorescence features

ABSTRACT We present a new analysis of the Suzaku X-ray spectrum of the Compton-thin Seyfert 2 galaxy NGC 4388. The spectrum above ∼2 keV can be described by a remarkably simple and rather mundane model, consisting of a uniform, neutral spherical distribution of matter, with a radial column density of $2.58\pm 0.02 \times 10^{23} \ \rm cm^{-2}$, and an Fe abundance of $1.102^{+0.024}_{-0.021}$ relative to solar. The model does not require any phenomenological adjustments to self-consistently account for the low-energy extinction, the Fe Kα and Fe Kβ fluorescent emission lines, the Fe K edge, and the Compton-scattered continuum from the obscuring material. The spherical geometry is not a unique description, however, and the self-consistent, solar abundance MYTORUS model, applied with toroidal and non-toroidal geometries, gives equally good descriptions of the data. In all cases, the key features of the spectrum are so tightly locked together that for a wide range of parameters, a relativistic disc-reflection component contributes no more than $\sim 2{{\ \rm per\ cent}}$ to the net spectrum in the 2–20 keV band. We show that the commonly invoked explanations for weak X-ray reflection features, namely a truncated and/or very highly ionized disc, do not work for NGC 4388. If relativistically broadened Fe Kα lines and reflection are ubiquitous in Seyfert 1 galaxies, they should also be ubiquitous in Compton-thin Seyfert 2 galaxies. The case of NGC 4388 shows the need for similar studies of more Compton-thin AGN to ascertain whether this is true.

On the cyclotron absorption line and evidence of the spectral transition in SMC X-2 during 2022 giant outburst

ABSTRACT We report comprehensive spectral and temporal properties of the Be/X-ray binary pulsar SMC X-2 using X-ray observations during the 2015 and 2022 outbursts. The pulse profile of the pulsar is unique and strongly luminosity dependent. It evolves from a broad-humped into a double-peaked profile above luminosity 3 × 1038 erg s−1. The pulse fraction of the pulsar is found to be a linear function of luminosity as well as energy. We also studied the spectral evolution of the source during the latest 2022 outburst with NICER. The observed photon index shows a negative and positive correlation below and above the critical luminosity, respectively, suggesting evidence of spectral transition from the sub-critical to supercritical regime. The broad-band spectroscopy of four sets of NuSTAR and XRT/NICER data from both outbursts can be described using a cut-off power-law model with a blackbody component. In addition to the 6.4 keV iron fluorescence line, an absorption-like feature is clearly detected in the spectra. The cyclotron line energy observed during the 2015 outburst is below 29.5 keV, however latest estimates in the 2022 outburst suggest a value of 31.5 keV. Moreover, an increase of 3.4 keV is detected in the cyclotron line energy at equal levels of luminosity observed in 2022 with respect to 2015. The observed cyclotron line energy variation is explored in terms of accretion induced screening mechanism or geometrical variation in line forming region.

Development of measures for metrological support of Raman spectroscopy

The method of Raman spectroscopy (RS) is widely used for timely metrological support of technological lines of the industrial sector in the chemical, medical and pharmaceutical, food, as well as criminalistics and forensic examinations. The wide application of the Raman spectroscopy method requires the use of specific metrological support tools, namely, measures for calibrating Raman spectrometers and microscopes according to the spectrum shape (i. e. relative spectral sensitivity).The purpose of the research was to develop prototype measures designed to calibrate Raman spectrometers and microscopes on a scale of relative intensities provided with metrological traceability to the SI base units.Prototype measures were made from inorganic glasses based on an oxide matrix, each of the glasses was activated with metal ions selected to excite a broad fluorescence line with radiation at a given wavelength: 532 nm (manganese ions), 633 nm (bismuth ions) and 785 nm (chromium ions). Metrological characteristics were established for prototype measures, where the certified characteristic is the relative intensity of the reproduced fluorescence radiation. The maximum expanded measurement uncertainty of the relative fluorescence intensity at a coverage factor k = 2 was determined, which is 9.4 %, 5.2 % and 2.8 % for prototype measures designed to reproduce the relative fluorescence intensity when excited at wavelengths of 532 nm, 633 nm and 785 nm, respectively.Certification of measures performed on the laser Raman confocal microscope Confotec NR500, which is part of the GET 196-2015 standard, allows establishing metrological traceability through the scale of relative intensities of the GET 8 6-2017 microscope, providing traceability to SI units of the “(light) energy flux” value. Thus, it is possible to find the spectral correction function for determining the Raman spectra traceable to the State Primary Standard GET 196-2015 for calibrated devices using certified measures.The practical significance of the results of the research makes it possible to expand the possibility of establishing and monitoring the stability of the calibration characteristics of microscopes and Raman spectrometers, namely, it allows calibration on a scale of relative intensities.

HABs Karenia brevis and Pseudo-nitzschia pre- and post-Hurricane Michael.

Increased occurrences of harmful algal blooms (HAB) in the Gulf of Mexico, and even worldwide, yield concern for increases in brevetoxin exposure leading to respiratory illness or even death, highlighting the need for extensive scientific research and human health monitoring. It is known that major events such as tropical storms and hurricanes are followed by periods of increased red tides caused by HABs; however, the nature by which phytoplankton blooms proliferate following major events remains a topic of great interest and research. The impact of Hurricane Michael on October 10, 2018 on HABs in the Florida panhandle was examined by analyzing data from the Florida Fish and Wildlife Conservation Commission in coordination with Normalized Fluorescence Line Height (nFLH) data from the University of South Florida College of Marine Science. Results presented here demonstrate four phases of HABs during storm events: 1. Pre-storm concentrations, 2. Decreased concentration during the storm, 3. Elevated concentrations following the storm and 4. Recovery period. This time frame can serve to be important in understanding the health dynamics of coastal systems following major storm events.

Exploring the Depths of Corrosion: A Novel GE-XANES Technique for Investigating Compositionally Complex Alloys.

In this study, we propose the use of nondestructive, depth-resolved, element-specific characterization using grazing exit X-ray absorption near-edge structure spectroscopy (GE-XANES) to investigate the corrosion process in compositionally complex alloys (CCAs). By combining grazing exit X-ray fluorescence spectroscopy (GE-XRF) geometry and a pnCCD detector, we provide a scanning-free, nondestructive, depth-resolved analysis in a sub-micrometer depth range, which is especially relevant for layered materials, such as corroded CCAs. Our setup allows for spatial and energy-resolved measurements and directly extracts the desired fluorescence line, free from scattering events and other overlapping lines. We demonstrate the potential of our approach on a compositionally complex CrCoNi alloy and a layered reference sample with known composition and specific layer thickness. Our findings indicate that this new GE-XANES approach has exciting opportunities for studying surface catalysis and corrosion processes in real-world materials.

Discovery of periodicities in two highly variable intermediate polars towards the Galactic centre

Aims. We performed a systematic analysis of X-ray point sources within 1.°5 of the Galactic centre using archival XMM-Newton data. While doing so, we discovered Fe Kα complex emission and pulsation in two highly variable sources, 4XMM J174917.7–283329 and 4XMM J174954.6–294336. In this work we report the findings of the X-ray spectral and timing studies. Methods. We performed detailed spectral modelling of the sources and searched for pulsation in the light curves using Fourier timing analysis. We also searched for multi-wavelength counterparts for the characterization of the sources. Results. The X-ray spectrum of 4XMM J174917.7–283329 shows the presence of complex Fe K emission in the 6–7 keV band. The equivalent widths of the 6.4 and 6.7 keV lines are 99−72+84 and 220−140+160 eV, respectively. The continuum is fitted by a partially absorbed apec model with a plasma temperature of kT = 13−2+10 keV. The inferred mass of the white dwarf (WD) is 0.9−0.2+0.3 M⊙. We detected pulsations with a period of 1212 ± 3 s and a pulsed fraction of 26 ± 6%. The light curves of 4XMM J174954.6–294336 display an asymmetric eclipse and dipping behaviour. To date, this is only the second known intermediate polar to show a total eclipse in X-rays. The spectrum of the sources is characterized by a power-law model with photon index Γ = 0.4 ± 0.2. The equivalent widths of the iron fluorescent (6.4 keV) and Fe XXV (6.7 keV) lines are 171−79+99 and 136−81+89 eV, respectively. The continuum is described by emission from optically thin plasma with a temperature of kT ∼ 35 keV. The inferred mass of the WD is 1.1−0.3+0.2 M⊙. We detect coherent pulsations from the source with a period of 1002 ± 2 s. The pulsed fraction is 66 ± 15%. Conclusions. The spectral modelling indicates the presence of intervening clouds with a high absorbing column density in front of both sources. The detected periodic modulations in the light curves are likely associated with the spin period of WDs in magnetic cataclysmic variables. The measured spin period, hard photon index, and equivalent width of the fluorescent Fe Kα line are consistent with the values found in intermediate polars. 4XMM J174954.6–294336 has already been classified as an intermediate polar, and we suggest that 4XMM J174917.7–283329 is a new intermediate polar. The X-ray eclipses in 4XMM J174954.6–294336 are most likely caused by a low-mass companion star obscuring the central X-ray source. The asymmetry in the eclipse is likely caused by a thick bulge that intercepts the line of sight during the ingress phase but not during the egress phase located behind the WD along the line of sight.

Evaluation of the clinical efficacy of quantitative light-induced fluorescence technology in diagnosing cracked teeth.

This retrospective study evaluated the clinical efficacy of quantitative light-induced fluorescence (QLF) technology for crack detection and the diagnosis of cracked teeth and assessed the possibility of a quantitative evaluation of cracks using QLF technology. Patients who were clinically diagnosed with cracked teeth over a 1-year period were included. The QLF images of the corresponding symptomatic cracked teeth and asymptomatic contralateral teeth with crack lines were taken with Qraypen C (AIOBIO, Seoul, Korea). Fluorescence loss (ΔF), maximum fluorescence loss (ΔFmax), red fluorescence (ΔR), and maximum red fluorescence (ΔRmax) of the crack line were analyzed. The correlation between these parameters and sex, age, tooth position (1st premolar, 2nd premolar, 1st molar, 2nd molar), spontaneous pain (+/-), percussion test (+/-), cold test (++/+/-), and bite test (+/-) were statistically analyzed. A total of 66 patients were included. Twenty-four patients had asymptomatic contralateral teeth with apparent crack lines; thus, 90 teeth were analyzed. The crack lines in 84 teeth observed as red fluorescent lines on the QLF images showed ΔR values higher than the cut-off value set by the analysis program used. The patient's age and the ∣ΔF∣ and ΔR values were positively correlated. However, there was no statistically significant difference in the QLF parameters between the same patient's symptomatic tooth and the contralateral tooth. QLF technology is a useful assistive diagnostic device for diagnosing cracked teeth.

Methodology and Results of Satellite Monitoring of Karenia Microalgae Blooms, That Caused the Ecological Disaster off Kamchatka Peninsula

The environmental disaster in Kamchatka in the autumn of 2020 was caused by an extensive bloom of harmful microalgae of the genus Karenia. A spectral shape algorithm was used to detect algae on satellite imagery. The algorithm calibration of in situ species composition data made it possible to identify areas where harmful algae dominated in biomass. The algorithm allowed evaluation of the dynamics of the distribution of the algae. The state of phytoplankton was estimated based on images of the specific capacity of photosynthesis. Specific fluorescence is the ratio of the height of the fluorescence line (flh) to the concentration of chlorophyll-a (chl-a). The parameter was used to recognize the stages of the algal bloom: intensive growth, blooming, and change in the dominant algal species. In addition, an increase in the concentration of harmful substances in the coastal zone due to wind impact was analyzed. After analyzing the available data, the events that caused the ecological disaster can be summarized as follows. After the stage of intensive growth of microalgae, nutrient deficiency stimulated the production of metabolites that have a harmful effect on the environment. The change of the dominant alga species in the second half of September and the past storm contributed to a sharp increase in the concentration of metabolites and dead organic matter in the coastal zone, which caused an ecological disaster. The subsequent mass bloom of alga species of the same genus, and the regular wind impact leading to the concentration of harmful substances in the coastal zone, contributed to the development of this catastrophic phenomenon.

Simultaneous 3D cellular positioning and apical dendritic morphology of transgenic fluorescent mouse CA3 hippocampal pyramidal neurons

BackgroundPyramidal neurons throughout hippocampal CA3 are diverse in their dendritic morphology, and CA3 is not homogenous in its structure or function. Nonetheless, few structural studies have captured the precise 3D somatic position and the 3D dendritic morphology of CA3 pyramidal neurons simultaneously. New methodHere, we present a simple approach to reconstruct the apical dendritic morphology of CA3 pyramidal neurons using the transgenic fluorescent Thy1-GFP-M line. The approach simultaneously tracks the dorsoventral, tangential, and radial positions of reconstructed neurons within the hippocampus. It is especially designed for use with transgenic fluorescent mouse lines, which are commonly used in genetic studies of neuronal morphology and development. ResultsWe demonstrate how topographic and morphological data are captured from transgenic fluorescent mouse CA3 pyramidal neurons. Comparison with existing methodsThere is no need to select and label CA3 pyramidal neurons with the transgenic fluorescent Thy1-GFP-M line. By taking transverse (not coronal) serial sections, we preserve fine dorsoventral, tangential, and radial somatic positioning of 3D-reconstructed neurons. Because CA2 is well defined by PCP4 immunohistochemistry, we use that technique here to to increase precision in defining tangential position along CA3. ConclusionsWe developed a method for simultaneously collecting precise somatic positioning as well as 3D morphological data among transgenic fluorescent mouse hippocampal pyramidal neurons. This fluorescent method should be compatible with many other transgenic fluorescent reporter lines and immunohistochemical methods, facilitating the capture of topographic and morphological data from a wide variety of genetic experiments in mouse hippocampus.

A dual-flow RootChip enables quantification of bi-directional calcium signaling in primary roots.

One sentence summary: Bi-directional-dual-flow-RootChip to track calcium signatures in Arabidopsis primary roots responding to osmotic stress. Plant growth and survival is fundamentally linked with the ability to detect and respond to abiotic and biotic factors. Cytosolic free calcium (Ca2+) is a key messenger in signal transduction pathways associated with a variety of stresses, including mechanical, osmotic stress and the plants' innate immune system. These stresses trigger an increase in cytosolic Ca2+ and thus initiate a signal transduction cascade, contributing to plant stress adaptation. Here we combine fluorescent G-CaMP3 Arabidopsis thaliana sensor lines to visualise Ca2+ signals in the primary root of 9-day old plants with an optimised dual-flow RootChip (dfRC). The enhanced polydimethylsiloxane (PDMS) bi-directional-dual-flow-RootChip (bi-dfRC) reported here adds two adjacent inlet channels at the base of the observation chamber, allowing independent or asymmetric chemical stimulation at either the root differentiation zone or tip. Observations confirm distinct early spatio-temporal patterns of salinity (sodium chloride, NaCl) and drought (polyethylene glycol, PEG)-induced Ca2+ signals throughout different cell types dependent on the first contact site. Furthermore, we show that the primary signal always dissociates away from initially stimulated cells. The observed early signaling events induced by NaCl and PEG are surprisingly complex and differ from long-term changes in cytosolic Ca2+ reported in roots. Bi-dfRC microfluidic devices will provide a novel approach to challenge plant roots with different conditions simultaneously, while observing bi-directionality of signals. Future applications include combining the bi-dfRC with H2O2 and redox sensor lines to test root systemic signaling responses to biotic and abiotic factors.

Changes in the distribution of circum-binary material around the HMXB GX 301-2 during a rapid spin-up episode of the neutron star

ABSTRACT Some accretion-powered X-ray pulsars with supergiant companion stars undergo occasional rapid spin-up episodes that last for weeks to a few months. We explore the changes in the accretion environment of the pulsar GX 301-2 during its latest 80 days long spin-up episode in 2019 when the spin frequency of the pulsar increased by ∼2 per cent over two orbits of the binary. By performing time-resolved spectroscopy with the MAXI/GSC spectra of the source, we estimated the equivalent hydrogen column density and equivalent width of the iron fluorescence line during the spin-up episode, and compared them with the long-term average values estimated by orbital-phase-resolved spectroscopy. The measured absorption column density during the spin-up episode is about twice that of an average orbit, while the equivalent width of the iron line is less than half of an average orbit. Though the spin-up episode started immediately after a pre-periastron flare and lasted for the two consecutive orbits of the binary, the associated enhancement in luminosity started a few days after the pre-periastron flare and lasted only during the first orbit, and some enhancement was seen again during the pre-periastron passage of the second orbit. The absorption column density and iron line equivalent width vary throughout the spin-up episode and are distinct from an average orbit. These observations indicate a significant change in the accretion and reprocessing environment in GX 301-2 during the spin-up episode and may hold important clues for the phenomenon in this source and several other sources with supergiant companions.

Generation of a human embryonic stem cell line (SKLRMe004-A) carrying NKX2.5-EGFP and TBX5-Tdtomato dual fluorescent reporters

TBX5 and NKX2-5 are two important transcription factors regulating cardiomyocytes commitment. In this study, we generated a dual fluorescent reporter cell line based on H9 human embryonic stem cells (hESCs) and called it H9-NKX2.5GFP/TBX5Td-T2A-18, which could be short for T2A-18. When T2A-18 was induced towards a cardiomyocyte fate, EGFP and Tdtomato fluorescences were observed, indicating the expression of NKX2.5 and TBX5, respectively. Meanwhile, T2A had normal karyotype and could maintain main characteristics of wildtype H9. Therefore, T2A-18 could be used as a tool to help us study the mechanism of cardiomyocytes specification from hESCs.

Split-GFP lamin as a tool for studying C. elegans LMN-1 dynamics in vivo.

We engineered a fluorescent fusion protein of C. elegans lamin, by fusing the eleventh beta strand of GFP to the N-terminus of LMN-1 at the endogenous lmn-1 locus. When co-expressed with GFP1-10, GFP11::LMN-1 was observed at the nuclear periphery of a wide variety of somatic cells. Homozygous gfp11::lmn-1 animals had normal numbers of viable embryos. However, the gfp11::lmn-1 animals had a mild swimming defect. While not completely functional, the GFP11::LMN-1 strain is more healthy than other published fluorescent LMN-1 lines, making it a valuable reagent for studying lamins.

Photosynthetic Fluorescence from Earthlike Planets around Sunlike and Cool Stars

Remote sensing of the Earth has demonstrated that photosynthesis is traceable as the vegetation red edge (VRE), which is a steep rise in the reflection spectrum of vegetation, and as solar-induced fluorescence. This study examines the detectability of biological fluorescence from two types of photosynthetic pigments, chlorophylls (Chls) and bacteriochlorophylls (BChls), on Earthlike planets with oxygen-rich/poor and anoxic atmospheres around the Sun and M dwarfs. Atmospheric absorption, such as H2O, CH4, O2, and O3, and the VRE obscure the fluorescence emissions from Chls and BChls. We find that the BChl-based fluorescence for wavelengths of 1000–1100 nm, assuming the spectrum of BChl b–bearing purple bacteria, could provide a suitable biosignature, but only in the absence of water cloud coverage or other strong absorbers near 1000 nm. The Chl fluorescence is weaker for several reasons, e.g., spectral blending with the VRE. The apparent reflectance excess is greatly increased in both the Chl and BChl cases around TRAPPIST-1, due to the fluorescence and stellar absorption lines. This could be a promising feature for detecting the fluorescence around ultracool red dwarfs using follow-up ground-based observations at high spectral resolution; however, this would require a long time around Sunlike stars, even for a LUVOIR-like space mission. Moreover, the simultaneous detection of fluorescence and the VRE is the key to identifying traces of photosynthesis, because absorption, reflectance, and fluorescence are physically connected. For further validation of the fluorescence detection, the nonlinear response of biological fluorescence as a function of light intensity could be considered.