Fluorescence Pattern Research Articles

The Discovery of the Hepatitis D Virus: Three Princes of Serendip and the Recognition of Autoantibodies to Liver-Kidney Microsomes.

The Discovery of the Hepatitis D Virus: Three Princes of Serendip and the Recognition of Autoantibodies to Liver-Kidney Microsomes.

Direct Optical Patterning of Quantum Dot Light-Emitting Diodes via In Situ Ligand Exchange.

Precise patterning of quantum dot (QD) layers is an important prerequisite for fabricating QD light-emitting diode (QLED) displays and other optoelectronic devices. However, conventional patterning methods cannot simultaneously meet the stringent requirements of resolution, throughput, and uniformity of the pattern profile while maintaining a high photoluminescence quantum yield (PLQY) of the patterned QD layers. Here, a specially designed nanocrystal ink is introduced, "photopatternable emissive nanocrystals" (PENs), which satisfies these requirements. Photoacid generators in the PEN inks allow photoresist-free, high-resolution optical patterning of QDs through photochemical reactions and in situ ligand exchange in QD films. Various fluorescence and electroluminescence patterns with a feature size down to ≈1.5 µm are demonstrated using red, green, and blue PEN inks. The patterned QD films maintain ≈75% of original PLQY and the electroluminescence characteristics of the patterned QLEDs are comparable to thopse of non-patterned control devices. The patterning mechanism is elucidated by in-depth investigation of the photochemical transformations of the photoacid generators and changes in the optical properties of the QDs at each patterning step. This advanced patterning method provides a new way for additive manufacturing of integrated optoelectronic devices using colloidal QDs.

A novel and selective multi-emission chemiluminescence system for the quantification of deltamethrin in food samples

A paper-based analytical device (PADs) combined with a powerful chemiluminescence (CL) system was established for the determination of deltamethrin (DM), based on the enhancing effect of polyphosphate (PP) on graphene quantum dots (GQDs)-KMnO4 CL reaction. A possible mechanism for the obtained emission was proposed using the CL spectra, fluorescence and ultraviolet-visible patterns. The reaction of KMnO4 and GQDs can lead to generation of GQDs in excited state (GQDs*), which can emit at 490 nm. Interestingly, PP changes the CL mechanism and the main emitter becomes Mn2+ instead of GQDs leading to a strong emission at 695 nm. Furthermore, the obtained multi-emission CL system was examined for analytical applications. The initial experiments showed that the amplified CL emission of the GQDs-KMnO4 system was selectively quenched in the presence of trace levels of DM, probably due to its effective interaction with GQDs or reaction with KMnO4. This observation led to a facile, reliable and sensitive PADs-CL probe, developed for the determination of DM residue in food samples. Using this CL system and under the optimized experimental conditions, the generated signal is decreased by increasing DM concentration in the range of 0.3−10 μg mL-1 with limit of detection (LOD) of 0.15 μg mL-1.

Preparation of 1, 3, 6, 8-Pyrenesulfonic Acid Tetrasodium Salt Dye-Doped Silica Nanoparticles and Their Application in Water-Based Anti-Counterfeit Ink.

In order to improve the luminescent stability of water-based anti-counterfeit ink, a new fluorescent material is prepared by doping dye into silica nanoparticles. Water soluble anionic dye 1, 3, 6, 8-pyrenesulfonic acid sodium salt (PTSA) is selected as the dopant. In this work, PTSA is successfully trapped into silica nanoparticles (SiNPs) by the reverse microemulsion method using cationic polyelectrolyte poly (dimethyl diallyl ammonium chloride; PDADMAC) as a bridge. The UV absorption spectra, fluorescence emission spectra and fluorescent decay curves are used to describe the luminescent properties of the PTSA-doped silica nanoparticles (PTSA-SiNPs). In addition, the as-prepared PTSA-SiNPs and polyurethane waterborne emulsion are used to prepare water-based anti-counterfeit ink, and fluorescent patterns are successfully printed through screen-printing. The samples printed by the ink exhibit desirable fluorescence properties, heat stability, robust photostability, and a fluorescent anti-counterfeit effect, which makes the PTSA-SiNPs promising luminescent materials for anti-counterfeit applications.

Endoscopic visualization of cancer and dysplasia in patients with ulcerative colitis following sensitization with oral 5-aminolevulinic acid.

ObjectiveEarly diagnosis of colitis‐associated cancer and dysplasia through surveillance endoscopy is vital for patients with ulcerative colitis (UC). This study aimed to evaluate the efficacy of autofluorescence endoscopy (AFE) using 5‐aminolevulinic acid (ALA) and to investigate the fluorescence signal localization pattern following 5‐ALA administration in tumorous lesions diagnosed as colitis‐associated cancer and dysplasia. The sensitivity and specificity of tumorous lesions detected by white light endoscopy (WLE) with and without AFE were evaluated.MethodsOverall, 13 endoscopic procedures were performed in 11 patients with UC using WLE and AFE following the oral administration of 5‐ALA. The biopsied lesions detected via endoscopy and resected specimens from cases underwent colectomy were assessed histopathologically. The sensitivity and specificity of detecting tumorous lesions by WLE with and without AFE were evaluated.ResultsOf the 68 lesions detected and biopsied, 63 were detected via WLE, and five were detected via AFE alone. The sensitivity of detecting colitis‐associated cancer and dysplasia via WLE combined with AFE was 36.4%, and the specificity, positive predictive value and negative predictive value were 94.2%, 57.1%, and 87.5%, respectively. Tumorous lesions displayed three types of fluorescence patterns on AFE.ConclusionsAFE using 5‐ALA can detect colitis‐associated cancer and dysplasia in patients with long‐standing UC and lesions that could not be detected via WLE. The distinctive fluorescence patterns in lesions may permit qualitative diagnoses of colitis‐associated cancer and dysplasia.

Aqueous Angiography in Normal Canine Eyes.

PurposeTo conduct aqueous angiography (AA) using a clinically applicable technique in normal dogs and to compare findings to intravenous scleral angiography (SA).MethodsWe examined 10 canine cadaver eyes and 12 eyes from live normal dogs. A gravity-fed trocar system delivered 2% sodium fluorescein and 0.25% indocyanine green (ICG) intracamerally (IC) in cadaver eyes. In vivo AA was subsequently performed in one eye of each of the 12 dogs via IC bolus of ICG under sedation. The same 12 dogs received SA via intravenous ICG (mean ± SD) 10.7 ± 3.3 days later. Identical scleral sectors were imaged using a Spectralis confocal scanning laser ophthalmoscope.ResultsThe gravity-fed trocar system permitted visualization of the conventional aqueous humor outflow (CAHO) pathways in cadaver eyes, but not in vivo. Fluorescence was observed superonasally in four of the 10 cadaver eyes within 24.0 ± 3.6 seconds. A single IC bolus of ICG showed CAHO pathways in vivo, demonstrating sectoral outflow patterns in the superotemporal sclera in 10 of the 12 eyes within 35.0 ± 4.3 seconds; four of the 12 eyes exhibited pulsatile aqueous movement. SA exhibited fluorescence patterns comparable to AA with weak pulsatile aqueous humor outflow.ConclusionsAngiography (AA or SA) in dogs permits visualization of the CAHO pathway and its vascular components in vivo. AA may be a more useful modality to assess aqueous humor outflow.Translational RelevanceIntracameral AA has potential utility for evaluating CAHO in vivo in dogs, an important animal model species.

Orcein and Fluorescent Banding Analysis of Two Floral Types of Catharanthus Roseus L.

Two floral types of Catharanthus roseus L. viz. pink and white were studied through differential staining with orcein, CMA and DAPI for cytogenetical characterization and to assist towards updating their taxonomical status and evaluating chromosomal diversity between them. "Simple Chromocenter Type" of interphase nuclei was observed with some darkly stained small heterochromatic regions throughout the nuclei. Most of the prophase chromosomes of Catharanthus roseus (pink and white) were "Continuous Type" and a few were "Gradient Type". Although these two floral types possessed 16 metacentric chromosomes in somatic cells, they showed variation in fluorescent banding pattern considering the modification of GC- and AT-rich repetitive segments. Taking into account all the parameters of both the floral types of C. roseus showed strict symmetric karyotype as well as primitive nature. Therefore, the combined data of differential staining provide information to make comments on their chromosomal status with cytogenetical characterization and also create a baseline for future research.
 Dhaka Univ. J. Biol. Sci. 29(2): 219-227, 2020 (July)

Expanding the Limits of Computer-Assisted Sperm Analysis through the Development of Open Software.

Computer assisted sperm analysis (CASA) systems can reduce errors occurring in manual analysis. However, commercial CASA systems are frequently not applicable at the forefront of challenging research endeavors. The development of open source software may offer important solutions for researchers working in related areas. Here, we present an example of this, with the development of three new modules for the OpenCASA software (hosted at Github). The first is the Chemotactic Sperm Accumulation Module, a powerful tool for studying sperm chemotactic behavior, analyzing the sperm accumulation in the direct vicinity of the stimuli. This module was validated by comparing fish sperm accumulation, with or without the influence of an attractant. The analysis clearly indicated cell accumulation in the treatment group, while the distribution of sperm was random in the control group. The second is the Sperm Functionality Module, based on the ability to recognize five sperm subpopulations according to their fluorescence patterns associated with the plasma membrane and acrosomal status. The last module is the Sperm Concentration Module, which expands the utilities of OpenCASA. These last two modules were validated, using bull sperm, by comparing them with visual counting by an observer. A high level of correlation was achieved in almost all the data, and a good agreement between both methods was obtained. With these newly developed modules, OpenCASA is consolidated as a powerful free and open-source tool that allows different aspects of sperm quality to be evaluated, with many potential applications for researchers.

Morphological Alterations of CAD Cells Overexpressing AKT1.

CAD cells are neuronal cells used in studies of cell differentiation and in cellular models of neuropathology. When cultured in differentiation medium, CAD cells exhibit characteristics of mature neurons including the generation of action potential. In addition to being a central signaling kinase in cell survival, AKT1 plays important roles in the nervous system including neuroplasticity and this study examined the localization of exogenous AKT1 in CAD cells. Neuropeptides modulate many signal transduction pathways and melacortins are implicated in regulating growth factor signal transduction pathways, including the PI3K/AKT pathway. AKT1-DsReD was transfected into CAD cells that were stably expressing melanocortin 3-receptor-GFP (MC3R-GFP), a G-protein coupled receptor. The cells were imaged with confocal microscopy to determine the fluorescent protein localization patterns. AKT1-DsRed was predominantly localized in the cytoplasm and the nucleus. Further, expression of exogenous AKT1 in these cell lines led to morphological changes reminiscent of apoptosis. As expected, MC3R-GFP localized to the plasma membrane but it internalized upon cell stimulation with the cognate ligand. In limited areas of the plasma membrane, AKT1-DsRed and MC3R-GFP were colocalized. In conclusion, quantitative studies to understand the role of relative levels of AKT1 in determining cell survival are needed.

Pericentromere clustering in Tradescantia section Rhoeo involves self-associations of AT- and GC-rich heterochromatin fractions, is developmentally regulated, and increases during differentiation

A spectacular but poorly recognized nuclear repatterning is the association of heterochromatic domains during interphase. Using base-specific fluorescence and extended-depth-of-focus imaging, we show that the association of heterochromatic pericentromeres composed of AT- and GC-rich chromatin occurs on a large scale in cycling meiotic and somatic cells and during development in ring- and bivalent-forming Tradescantia spathacea (section Rhoeo) varieties. The mean number of pericentromere AT-rich domains per root meristem nucleus was ca. half the expected diploid number in both varieties, suggesting chromosome pairing via (peri)centromeric regions. Indeed, regular pairing of AT-rich domains was observed. The AT- and GC-rich associations in differentiated cells contributed to a significant reduction of the mean number of the corresponding foci per nucleus in relation to root meristem. Within the first 10 mm of the root, the pericentromere attraction was in progress, as if it was an active process and involved both AT- and GC-rich associations. Complying with Rabl arrangement, the pericentromeres preferentially located on one nuclear pole, clustered into diverse configurations. Among them, a strikingly regular one with 5–7 ring-arranged pericentromeric AT-rich domains may be potentially engaged in chromosome positioning during mitosis. The fluorescent pattern of pachytene meiocytes and somatic nuclei suggests the existence of a highly prescribed ring/chain type of chromocenter architecture with side-by-side arranged pericentromeric regions. The dynamics of pericentromere associations together with their non-random location within nuclei was compared with nuclear architecture in other organisms, including the widely explored Arabidopsis model.

Neuropatía periférica como manifestaciónunica de poliangeítis microscópica en la altura. Reporte de caso

La poliangeítis microscópica, es una vasculitis de vasos pequeños cuya incidencia es de 4 por 1 000 000 de habitantes y que frecuentemente se presenta asociada a manifestaciones de glomerulonefrítis rápidamente progresiva y capilaritis pulmonar. Las manifestaciones neurológicas son menos frecuentes. El diagnostico se confirma con serología positiva para Anticuerpos Anticitoplasma de neutrófilos (ANCA) y especificidad para patrón de fluorescencia perinuclear (P-ANCA) y Mieloperoxidasa (MPO). El tratamiento idóneo son los glucocorticoides e inmunosupresores. Se presenta el caso de un paciente de 69 años con poliangeitis microscópica que debuta con neuropatía periférica como única manifestación clínica, presentación que se considera atípica. Se plantea que ante la presencia de una neuropatía periférica aislada, deben efectuarse estudios para confirmar o descartar la presencia de Poliangeitis Microscópica en base a la gran variabilidad de su sintomatología clínica y potenciales formas de tratamiento.

NOSA, an Analytical Toolbox for Multicellular Optical Electrophysiology.

Understanding how neural networks generate activity patterns and communicate with each other requires monitoring the electrical activity from many neurons simultaneously. Perfectly suited tools for addressing this challenge are genetically encoded voltage indicators (GEVIs) because they can be targeted to specific cell types and optically report the electrical activity of individual, or populations of neurons. However, analyzing and interpreting the data from voltage imaging experiments is challenging because high recording speeds and properties of current GEVIs yield only low signal-to-noise ratios, making it necessary to apply specific analytical tools. Here, we present NOSA (Neuro-Optical Signal Analysis), a novel open source software designed for analyzing voltage imaging data and identifying temporal interactions between electrical activity patterns of different origin. In this work, we explain the challenges that arise during voltage imaging experiments and provide hands-on analytical solutions. We demonstrate how NOSA’s baseline fitting, filtering algorithms and movement correction can compensate for shifts in baseline fluorescence and extract electrical patterns from low signal-to-noise recordings. NOSA allows to efficiently identify oscillatory frequencies in electrical patterns, quantify neuronal response parameters and moreover provides an option for analyzing simultaneously recorded optical and electrical data derived from patch-clamp or other electrode-based recordings. To identify temporal relations between electrical activity patterns we implemented different options to perform cross correlation analysis, demonstrating their utility during voltage imaging in Drosophila and mice. All features combined, NOSA will facilitate the first steps into using GEVIs and help to realize their full potential for revealing cell-type specific connectivity and functional interactions.

Cocaïne et lésions destructrices centro-faciales : à propos d'un cas

IntroductionCocaine use is associated with multiple complications, some of which can mimic systemic diseases, especially Antineutrophil Cytoplasmic Antibody (ANCA) associated vasculitis. We report a case of Cocaine Induced Midline Destructive Lesions (CIMDL) for which a diagnosis of granulomatosis with polyangiitis (GPA) was discussed. Case reportA 42-year-old male, cocaine consumer, was admitted in our department for a centrofacial destructive process. He had no extra ear, nose and throat (ENT) involvement. ANCA were positive with a perinuclear fluorescence pattern and an anti-Proteinase 3 specificity. Regarding this unusual immunologic pattern and in the absence of histological argument for a GPA, a diagnosis of CIMDL was made. ConclusionCIMDL is a centrofacial destructive process due to intranasal cocaine use. It is frequently associated with the presence of p-ANCA with both anti-HNE and anti-PR3 specificity.

Synthesis and Docking Study of N-(Cinnamoyl)-N'-(substituted)acryloyl Hydrazide Derivatives Containing Pyridinium Moieties as a Novel Class of Filamentous Temperature-Sensitive Protein Z Inhibitors against the Intractable Xanthomonas oryzae pv. oryzae Infections in Rice.

Xanthomonas oryzae pv. oryzae (Xoo) is an offensive phytopathogen that can invade a wide range of plant hosts to develop bacterial diseases, including the well-known rice bacterial leaf blight. However, few agrochemicals have been identified to effectively prevent and eliminate Xoo-induced diseases. Thus, designing novel antibacterial compounds on the basis of the potential targets from Xoo may lead to the discovery of highly efficient and innovative anti-Xoo agents. Filamentous temperature-sensitive protein Z (FtsZ), an important functional protein in the progression of cell division, has been widely reported and exploited as a target for creating antibacterial drugs in the field of medicine. Therefore, the fabrication of innovative frameworks targeting XooFtsZ may be an effective method for managing bacterial leaf blight diseases via blocking the binary division and reproduction of Xoo. As such, a series of novel N-(cinnamoyl)-N'-(substituted)acryloyl hydrazide derivatives containing pyridinium moieties were designed, and the anti-Xoo activity was determined. The bioassay results showed that compound A7 had excellent anti-Xoo activity (EC50 = 0.99 mg L-1) in vitro and distinct curative activity (63.2% at 200 mg L-1) in vivo. Further studies revealed that these designed compounds were XooFtsZ inhibitors, validating by the reduced GTPase activity of recombinant XooFtsZ, the nonfilamentous XooFtsZ assembly observed in the TEM images, and the prolonged Xoo cells from the fluorescence patterns. Computational docking studies showed that compound A7 had strong interactions with ASN34, GLN193, and GLN197 residues located in the α helix regions of XooFtsZ. The present study demonstrates the developed FtsZ inhibitors can serve as agents to control Xoo-induced infections.

Lupus 2020

The publication of the new ACR/EULAR classification criteria for systemic lupus erythematosus (SLE) and the revision of the EULAR recommendations for the management of SLE last year include many important novelties that will change the classification and care of lupus patients. This refers to both general basic treatment principles and specific therapy recommendations. For example, SLE can only be classified as such if antinuclear antibodies (ANA) are present, independently of titer height and fluorescence pattern. If ANAs are detected, lupus-specific symptoms and serological abnormalities are assessed and separately scored. A score of 10 and more classifies an SLE patient as such. With regard to the care of SLE patients, every therapeutic decision should follow a treat-to-target approach with the aim of achieving remission. Aim is to ensure long-term survival, reduce damage and improve quality of life. These goals are achieved by early diagnosis, consistent therapy and regular evaluation of disease activity. All patients should receive antimalarials in the absence of contraindications. If the response is inadequate or in case of moderate to severe organ manifestations, steroid pulses over a few days and the early use of immunosuppressive drugs (IM) are recommended. With the inclusion of biologicals, especially belimumab and rituximab, new substance classes made their way into the recommendations.

Seasonal patterns of near-bottom chlorophyll fluorescence in the eastern Chukchi Sea: 2010–2019

The Chukchi Sea consists of a broad, shallow (<45 m) shelf that is seasonally (November–July) covered by sea ice. This study characterizes the seasonal patterns of near-bottom primary production using moored instruments measuring chlorophyll fluorescence, oxygen, nitrate, and photosynthetically active radiation. From 2010 to 2018, moorings were deployed at multiple sites each year. Instruments were restricted to within 10 m of the seafloor due to ice keels, which can reach 30 m below the surface in this region. Near-bottom blooms were common at all mooring sites. The bloom onset directly followed ice retreat whereas the end of the bloom followed loss of light in September. The intensity of light at the seafloor (~40 m deep) was similar to levels observed under 1–2 m thick ice floes in the spring/early summer, and was sufficient to support photosynthesis near the seafloor, utilizing nitrate and producing oxygen. We hypothesize that the near bottom bloom originated from aggregates of ice algae that sank during ice retreat. As a consequence of climate warming and earlier ice retreat, we predict that the near-bottom bloom onset will occur earlier, but the timing of the end of the near-bottom bloom will remain the same pending a sufficient nutrient supply. The Chukchi Sea is highly productive even though the growing season is short. This production is promoted by a shallow seafloor, which allows multiple production layers (surface open water, bottom of the mixed layer, under-ice algae, and disassociated ice algae which settles near the seafloor). We term this the Multiple Production Layers (MPL) hypothesis.

Pathological Validity of Using Near-infrared Fluorescence Imaging for Securing Surgical Margins During Liver Resection.

This study investigated the use of near-infrared fluorescent imaging for securing safe margins during liver resection. This study included 125 patients who underwent liver tumor resection in 2014-2018. Indocyanine green testing was performed 2-14 days before surgery. Histopathological specimens of hepatocellular carcinoma (HCC) and colorectal liver metastasis (CRLM) were evaluated using fluorescent microscopy. Fluorescence microscopy identified signals in 26/53 (49.0%) and 36/72 (50%) cases of HCC and CRLM, respectively. HCC demonstrated total, partial, rim, and combined fluorescence patterns; CRLM uniformly demonstrated rim fluorescence. Although rim fluorescence was seen in both HCC and CRLM, no malignancy was confirmed pathologically in the peritumoral area demonstrating fluorescence. The median widths of fluorescence from the tumor edge in HCC and CRLM were 1227.5 μm and 1608 μm, respectively, with no significant difference. Near-infrared fluorescent imaging can reliably detect safe surgical margins intraoperatively during liver resection.

Naphthalimide‐Based Aggregation‐Induced Emissive Polymeric Hydrogels for Fluorescent Pattern Switch and Biomimetic Actuators

Front Cover: In article 2000123, Wei Lu, Tao Chen and co-workers prepare a robust aggregation-induced emissive polymeric hydrogel by copolymerization of N-isopropylacrylamide and a specially designed 4-phenoxy-N-allyl-1,8-naphthalimide (PhAN) monomer. The hydrogel is characterized with thermally controlled emission/transmittance response, and its potential applications as a fluorescence pattern switch and biomimetic actuator are demonstrated.

Dermatofluoroscopy Is Also for Redheads a Sensitive Method of Early Melanoma Detection

Background: Caucasians with red hair and fair skin have a remarkably increased risk of malignant melanoma compared to non-redhead Caucasians. Objectives: With the aim of a reliable melanoma diagnosis in redheads, the application of dermatofluoroscopy was analyzed in 16 patients with red hair. Most of them had been included in a clinical dermatofluoroscopy study for patients with the suspicion of melanoma. We examined whether the 25 lesions of the redheads showed the same characteristic melanin fluorescence spectra for dysplastic nevi and melanomas as those of non-redhead Caucasians or whether there was a different fluorescence pattern. This is important in view of the known significantly altered ratio of eumelanin to pheomelanin in the skin of redheads. Methods: More than 8,000 spatially resolved fluorescence spectra of 25 pigmented lesions were measured and analysed. The spectra were excited by the stepwise absorption of two 800-nm photons (principle of dermatofluoroscopy). Furthermore, the fluorescence spectra of eumelanin and pheomelanin in hair samples were determined in the same way. Results: The evaluation revealed that the melanin fluorescence spectra of dysplastic nevi and melanomas of redheads have the same spectral characteristics as those of non-redhead Caucasians. An accompanying result is that dermatofluoroscopy shows identical fluorescence spectra for eumelanin and pheomelanin. Conclusions: Dermatofluoroscopy proves to be a reliable diagnostic method also for redheads. Our results also explain our recent finding that there is a uniform fluorescence spectroscopic fingerprint for melanomas of all subtypes, which is of particular interest for hypomelanotic and apparently amelanotic melanomas containing pheomelanin.

Exploiting geometric similarity for statistical quantification of fluorescence spatial patterns in bacterial colonies

BackgroundCurrently the combination of molecular tools, imaging techniques and analysis software offer the possibility of studying gene activity through the use of fluorescent reporters and infer its distribution within complex biological three-dimensional structures. For example, the use of Confocal Scanning Laser Microscopy (CSLM) is a regularly-used approach to visually inspect the spatial distribution of a fluorescent signal. Although a plethora of generalist imaging software is available to analyze experimental pictures, the development of tailor-made software for every specific problem is still the most straightforward approach to perform the best possible image analysis. In this manuscript, we focused on developing a simple methodology to satisfy one particular need: automated processing and analysis of CSLM image stacks to generate 3D fluorescence profiles showing the average distribution detected in bacterial colonies grown in different experimental conditions for comparison purposes.ResultsThe presented method processes batches of CSLM stacks containing three-dimensional images of an arbitrary number of colonies. Quasi-circular colonies are identified, filtered and projected onto a normalized orthogonal coordinate system, where a numerical interpolation is performed to obtain fluorescence values within a spatially fixed grid. A statistically representative three-dimensional fluorescent pattern is then generated from this data, allowing for standardized fluorescence analysis regardless of variability in colony size. The proposed methodology was evaluated by analyzing fluorescence from GFP expression subject to regulation by a stress-inducible promoter.ConclusionsThis method provides a statistically reliable spatial distribution profile of fluorescence detected in analyzed samples, helping the researcher to establish general correlations between gene expression and spatial allocation under differential experimental regimes. The described methodology was coded into a MATLAB script and shared under an open source license to make it accessible to the whole community.