Comparison of residual silicone oil index after removal of silicone oil with fluid-air versus oil-fluid exchange.

Objectives: To compare the effectiveness of fluid-air exchange with silicone oil-fluid exchange in reducing the residual silicone oil (SO) droplets after the removal of SO. Methods: This was a prospective, quasi-experimental study conducted from October 2021 to February 2022 at Eye Unit-III, COAVS, Mayo Hospital, Lahore. Sixty-one patients with siliconized eyes underwent removal of SO with two different techniques and were divided into fluid-air exchange and oil-fluid exchange groups. To quantify the residual silicone droplets objectively, B-scan echographic images were analyzed within seven days of surgery. Silicone oil index (SOI) which is the amount of residual SO droplets/vitreal area in the images was calculated with the help of imagej software. Results: The residual SOI of the fluid-air exchange group (0.99 ± 1.76%) was significantly lower than the oil-fluid exchange group (3.25 ± 3.85%). The SOI is positively correlated with the duration of tamponade, preoperative intraocular- pressure and axial length. Persistent IOP elevation post-operatively was seen in 16.67% individuals in the fluid-air exchange group and 54.8% individuals in the oil-fluid exchange group. Conclusion: Fluid-air exchange group was found to be superior in reducing residual SO droplets than the oil-fluid exchange group.

Changes in the tear film and meibomian gland morphology between preclinical dry eye and normal subjects represented by ocular surface disease index scores.

Tear film and MG morphology play an essential role in detecting dry eyes and Meibomian Gland Dysfunction (MGD). Previous studies have reported these changes in cases diagnosed with dry eyes. However, it is important to study these changes among subjects with symptoms of dry eyes without signs. Hence, this study evaluated the changes in the tear film and meibomian gland (MG) morphology among subjects with pre-clinical dry eye and compared it with the normals defined by the Ocular Surface Disease Index score. One hundred and fifty subjects were enrolled in this prospective cross-sectional study. All the subjects completed the Ocular Surface Disease Index (OSDI) questionnaire and the Computer Vision Syndrome Questionnaire (CVS-Q). Tear film tests such as non-invasive break-up time (NIBUT), tear meniscus height (TMH), lipid layer pattern, Schirmer's test, and corneal staining were performed. Images were captured from both the upper and the lower eyelids to study the MG morphology. TMH, MG length, thickness, loss, and tortuosity were measured using ImageJ software. Subjects were differentiated into two groups based on an OSDI threshold score of ≥13 as preclinical dry eye and normals. Among all the subjects, 43.6% of them were categorized as pre-clinical dry eye, and 56.4% as normals. In the pre-clinical dry eye group, a significant reduction in NIBUT (Z = -2.13, p = 0.03) and MG length of the lower lid (Z = -2.58, p = 0.01) was found when compared to normals. TMH, Schirmer's test, and MG width did not vary among both groups (p > 0.05). Similarly, MG loss and tortuosity score was higher in the pre-clinical dry eye group, but did not show any statistical significance (p > 0.05). The majority of the young individuals presenting for the regular eye examination may have pre-clinical dry eye based on their OSDI scores. Hence, it is important to administer the OSDI questionnaire and perform non-invasive tests such as NIBUT and IR imaging of MG as a part of the routine eye examination.

Impact of using magnetic water on the micro structure of leached saline-sodic soil.

Structural changes in the porous media are critical in evaluating the soil pore system and other physical properties of leached soil. In this work, the combination of selected physical parameters such as bulk density and hydraulic conductivity with images of scanning electron microscopic (SEM) was used to study the impact of five levels (1, 3, 5, 7, and 9 magnets) of magnetically treated water (MTW) on the structural changes in pore spaces of leached high saline-sodic soil, compared with leached soil with non- magnetically treated water (NMTW) and unleached soil. Results show that leaching saline-sodic soil with MTW leads to a remarkable increase in soil pore network as confirmed by SEM observations and ImageJ software program. The influence of MTW is important in all cases but the changes in pores of leached soil were mainly observed when the magnetic fields of 7 and 9 were used. It has been found that the value of total percentage area of pores in the leached soil was found to be 7.55 and 9.53, respectively, while the total percentage area of pores in both leached soil with NMTW and unleached soil were 0.69 and 0.1 respectively. It can be concluded that MTW can improve the physiochemical properties of soils in two ways: first, it accelerates the solubility of salts and increases calcium ion available on the soil exchange surface which displaced a higher concentration of adsorbed sodium ions. Second, the changes in the microstructure lead to improved soil porosity combined with saturated hydraulic conductivity and bulk density. This study will provide a method for using magnetic water technology to improve soil properties greatly by increasing soil porosity. Through this porosity, water, solutes, and gases can diffuse through the ground, thus increasing exchange processes.

Transient receptor potential vanilloid 4 activation inhibits the delayed rectifier potassium channels in hippocampal pyramidal neurons: An implication in pathological changes following pilocarpine-induced status epilepticus.

Activation of transient receptor potential vanilloid 4 (TRPV4) can increase hippocampal neuronal excitability. TRPV4 has been reported to be involved in the pathogenesis of epilepsy. Voltage-gated potassium channels (VGPCs) play an important role in regulating neuronal excitability and abnormal VGPCs expression or function is related to epilepsy. Here, we examined the effect of TRPV4 activation on the delayed rectifier potassium current (IK ) in hippocampal pyramidal neurons and on the Kv subunits expression in male mice. We also explored the role of TRPV4 in changes in Kv subunits expression in male mice following pilocarpine-induced status epilepticus (PISE). Application of TRPV4 agonists, GSK1016790A and 5,6-EET, markedly reduced IK in hippocampal pyramidal neurons and shifted the voltage-dependent inactivation curve to the hyperpolarizing direction. GSK1016790A- and 5,6-EET-induced inhibition of IK was blocked by TRPV4 specific antagonists, HC-067047 and RN1734. GSK1016790A-induced inhibition of IK was markedly attenuated by calcium/calmodulin-dependent kinase II (CaMKII) antagonist. Application of GSK1016790A for up to 1 hr did not change the hippocampal protein levels of Kv1.1, Kv1.2, or Kv2.1. Intracerebroventricular injection of GSK1016790A for 3 d reduced the hippocampal protein levels of Kv1.2 and Kv2.1, leaving that of Kv1.1 unchanged. Kv1.2 and Kv2.1 protein levels as well as IK reduced markedly in hippocampi on day 3 post PISE, which was significantly reversed by HC-067047. We conclude that activation of TRPV4 inhibits IK in hippocampal pyramidal neurons, possibly by activating CaMKII. TRPV4-induced decrease in Kv1.2 and Kv2.1 expression and IK may be involved in the pathological changes following PISE.

Smartphone coupled with paper-based chemical sensor for on-site determination of iron(III) in environmental and biological samples.

We report a smartphone-paper-based sensor impregnated with cetyltrimethylammonium bromide modified silver nanoparticles (AgNPs/CTAB) for determination of Fe3+ in water and blood plasma samples. The methodology for determination of Fe3+ is based on the change in signal intensity of AgNPs/CTAB fabricated on a paper substrate after the deposition of analyte, using a smartphone followed by processing with ImageJ software. The mechanism of sensing for detection and determination of Fe3+ is based on the discoloration of AgNPs which impregnated the paper substrate. The discoloration is attributed to the electron transfer reaction taking place on the surface of NPs in the presence of CTAB. Fe3+ was determined when the paper was impregnated with 1 mM AgNPs for 5 min of reaction time and the substrate was kept under acidic conditions. The linear range for determination of total iron in terms of Fe3+ was 50-900 µg L-1 with a limit of determination (LOD) of 20 µg L-1 and coefficient of variation (CV) of 3.2%. The good relative recovery of 91.3-95.0% and interference studies showed the selectivity of the method for determination of total iron in water and blood plasma samples. Smartphone-paper-based sensors have advantages of simplicity, rapidity, user-friendliness, low cost, and miniaturization of the method for on-site determination of total iron compared to methods that require sophisticated analytical instruments. Graphical abstract Smartphone-paper-based sensor with cetyltrimethylammonium bromide modified silver nanoparticles for determination of Fe3+ in water and blood plasma samples.

Combining qualitative and quantitative imaging evaluation for the assessment of genomic DNA integrity: The SPIDIA experience.

In this note, we present an ad hoc procedure that combines qualitative (visual evaluation) and quantitative (ImageJ software) evaluations of Pulsed-Field Gel Electrophoresis (PFGE) images to assess the genomic DNA (gDNA) integrity of analyzed samples. This procedure could be suitable for the analysis of a large number of images by taking into consideration both the expertise of researchers and the objectiveness of the software. We applied this procedure on the first SPIDIA DNA External Quality Assessment (EQA) samples. Results show that the classification obtained by this ad hoc procedure allows a more accurate evaluation of gDNA integrity with respect to a single approach.

TreeJ: an ImageJ plugin for interactive cell lineage reconstruction from static images.

BACKGROUND: With the emergence of deep-learning methods, tools are needed to capture and standardize image annotations made by experimentalists. In developmental biology, cell lineages are generally reconstructed from time-lapse data. However, some tissues need to be fixed to be accessible or to improve the staining. In this case, classical software do not offer the possibility of generating any lineage. Because of their rigid cell walls, plants present the advantage of keeping traces of the cell division history over successive generations in the cell patterns. To record this information despite having only a static image, dedicated tools are required. RESULTS: We developed an interface to assist users in the building and editing of a lineage tree from a 3D labeled image. Each cell within the tree can be tagged. From the created tree, cells of a sub-tree or cells sharing the same tag can be extracted. The tree can be exported in a format compatible with dedicated software for advanced graph visualization and manipulation. CONCLUSIONS: The TreeJ plugin for ImageJ/Fiji allows the user to generate and manipulate a lineage tree structure. The tree is compatible with other software to analyze the tree organization at the graphical level and at the cell pattern level. The code source is available at https://github.com/L-EL/TreeJ .

Image calibration in choroidal vascularity index measurement.

Choroid is a tissue with a very high blood flow which is a metabolic supporter of the retina. Recently, the study of choroidal blood flow in ocular and systemic disorders is a hot topic in scientific research. With the advent of enhanced depth imaging OCT (EDI-OCT), it is possible to measure the entire choroidal thickness. The choroidal vascularity index (CVI) is a relatively new index in studying choroidal hemodynamics. However, the CVI measurement needs image processing. Image calibration is a necessary step before any image processing with software such as ImageJ.

Customization of spectacle frames for individuals with facial deformities by altering conventional spectacle frames - Phase I.

Purpose: Manufacturing a spectacle frame for a facially deformed individual is challenging because of facial asymmetry. One of the solutions is the customization of spectacle frames. Customization of spectacle frames for facially deformed individuals requires a better understanding of the facial anthropometry of deformed faces. This study aimed to analyze the facial anthropometry of deformed faces to understand the range of variability. The results of this study will be used to find customization methods in the future. Methods: We measured and analyzed the 12 facial parameters of individuals with facial deformities using the ImageJ software. Results: The data collected were normally distributed. Paired sample test revealed a statistically significant difference between innercanthus distances (right innercanthus distance [RICD] and left innercanthus distance [LICD]). Correlation analysis showed a positive difference between horizontal and vertical pupillary distance-innercanthus distance (PD-ICD) (0.68, 0.75, and 0.81) and pupillary distance-helix distance (PD-HD) (0.57, 0.68, and 0.59) relations. PD-ICD correlations are stronger compared to the PD-HD relation. Conclusion: Altering the frame center distance and the temple heights are the most important for asymmetric faces. Large population-based data are required to make concrete decisions to design a spectacle frame for asymmetric faces.

Comparison of single catheter versus dual catheter-based EBT3 film calibration for the Ir-192 beam energy.

Purpose. Films and TLDs have been the common choices for passivein-vivodose measurement in radiotherapy. In the brachytherapy applications, it is very difficult to report and verify the dose at multiple localized high dose gradient regions and also the dose to organ at risk. This study was carried out to introduce a new and accurate calibration method for GafChromic EBT3 films irradiated using Ir-192 photon energy from miniature High Dose Rate (HDR) Brachytherapy source.Materials and methods. Film holder made of Styrofoam was used to hold the EBT3 film at its center. It was placed inside the mini water phantom and the films were irradiated by Ir-192 source of microSelectron HDR afterloading brachytherapy system. Two different setups: Single catheter-based film exposure and dual catheter-based film exposure were compared. The films scanned on a flatbed scanner were analysed in three different color channels: red, green, and blue using Image J software. The dose calibration graphs were generated using the third-order polynomial equations fitted on the data points from two different methods of calibration procedure. Maximum and mean dose difference between TPS calculated and measured was analyzed.Results. The measured dose difference from the TPS calculated doses were evaluated for the three groups of dose ranges (low, medium and high). In the high dose range, standard uncertainty of dose difference are ±2.3%, ±2.9%, and ±2.4% respectively for the red, green, and blue color channel when the TPS calculated dose was compared with single catheter based film calibration equation. Whereas it is observed as 1.3%, 1.4% and 3.1% for the red, green, and blue color channels respectively when compared with the dual catheter based film calibration equation. A test film was exposed to a TPS calculated dose of 666 cGy to validate the calibration equations, single catheter based film calibration equation estimated the dose difference as -9.2%, -7.8% and -3.6% respectively in the red, green, and blue color channels whereas the same were observed as 0.1%, 0.2% and 6.1% respectively when dual catheter based film calibration equation was applied.Conclusion.Source miniature size, reproducible positioning of the film and catheter system inside water medium are the major challenges in the film calibration with Ir-192 beam. To overcome these situations dual catheter-based film calibration was found more accurate and reproducible as compare to the single catheter based film calibration.

Characterization of the immune infiltrate in mouse tissue by multiplex immunofluorescence.

Multiplexed immunofluorescence imaging of formalin-fixed, paraffin-embedded (FFPE) specimens mounted on glass slides allow the identification of multiple cell phenotypes while retaining spatial and morphological context. Multiplex immunofluorescence protocols have already been developed and validated for mouse tissues. Immunophenotyping analysis reliably depicts the immune landscape of cancer tissues that has been demonstrated to influence cancer development and progression as well as to have an impact on therapy responsiveness and resistance. Here, we describe a method for multiplexed fluorescence image analysis, enabling analysis of mouse cancer morphology and cell phenotypes in FFPE sections.

Automated, image-based disease measurement for phenotyping resistance to soybean frogeye leaf spot.

BACKGROUND: Frogeye leaf spot is a disease of soybean, and there are limited sources of crop genetic resistance. Accurate quantification of resistance is necessary for the discovery of novel resistance sources, which can be accelerated by using a low-cost and easy-to-use image analysis system to phenotype the disease. The objective herein was to develop an automated image analysis phenotyping pipeline to measure and count frogeye leaf spot lesions on soybean leaves with high precision and resolution while ensuring data integrity. RESULTS: The image analysis program developed measures two traits: the percent of diseased leaf area and the number of lesions on a leaf. Percent of diseased leaf area is calculated by dividing the number of diseased pixels by the total number of leaf pixels, which are segmented through a series of color space transformations and pixel value thresholding. Lesion number is determined by counting the number of objects remaining in the image when the lesions are segmented. Automated measurement of the percent of diseased leaf area deviates from the manually measured value by less than 0.05% on average. Automatic lesion counting deviates by an average of 1.6 lesions from the manually counted value. The proposed method is highly correlated with a conventional method using a 1-5 ordinal scale based on a standard area diagram. Input image compression was optimal at a resolution of 1500 × 1000 pixels. At this resolution, the image analysis method proposed can process an image in less than 10 s and is highly concordant with uncompressed images. CONCLUSION: Image analysis provides improved resolution over conventional methods of frogeye leaf spot disease phenotyping. This method can improve the precision and resolution of phenotyping frogeye leaf spot, which can be used in genetic mapping to identify QTLs for crop genetic resistance and in breeding efforts for resistance to the disease.

Application of ImageJ Software for the Quantification of Hand Joint Space Narrowing in Patients with Rheumatoid Arthritis.

BACKGROUND: ImageJ software is used to quantify the joint space width (JSW) of hand and wrist in patients with rheumatoid arthritis (RA) as well as in the healthy control group. METHODS: Forty-one RA patients and 31 healthy controls were included in this study. All of 72 participants underwent digital radiography of the bilateral hand and wrist; then, all the images were opened by ImageJ software to measure the width of wrist and hand joint space (total 2160 joints). Joint space narrowing (JSN) was defined if the width was less than the mean - 2SD of the control group. RESULT: The mean JSW of all sites of wrist and hand joints of RA patients were significantly reduced as compared to those in the control group (p<0.001). There were 37/41 (90.24%) RA patients who had JSN in at least one joint in hand or wrist. In total, 70.89% of joints on the right and 68.46% of joints on the left wrist and hand had JSN. CONCLUSION: ImageJ software is simple and convenient that helps rheumatologists quantify the width of joint space for diagnosis and follow-up in RA patients.

Bioinformatic Analysis of MIRU-VNTR Repeats of Mycobacterium Tuberculosis Strains from Lagos, Nigeria.

The analysis of Mycobacterial Interpersed Repetitive Unit-Variable Number of Tandem Repeat (MIRU-VNTR) discriminates against the species of M. tuberculosis involved in the transmission of the disease. The reference method is the manual method. Our study involved developing a bioinformatics method of interpreting MIRU-VNTR and comparing it to the manual method. For this we used two softwares, namely imagej and Microsoft Excel. Imagej was used to determine the migration distance of the bands and for the measure of size in a base pair. The number of repetitions of 18 markers used was analyzed with Excel macro. The results obtained were: 27% of the results exactly consistent, 16% of outliers generated by the macro and 57% of the results not matching.

Determination of Collimator Helmet Factors for Leksell Gamma Knife 4C Unit Using GAF Chromic EBT3 Film and ImageJ Software.

BACKGROUND: Measurement of Collimator helmet factors (CHF) is an important quality assurance procedure to be performed on Leksell Gamma Knife unit at regular interval to make sure that the interchangeable collimator helmet fit into the source channels without any positional inaccuracy which leads to major treatment error. The primary aim of this study is to measure the CHFs for Elekta Leksell Gamma knife 4C helmets using GafChromic EBT3 film and Image J software. METHODS: GafChromic EBT3 film, EPSON expression 10000 XL scanner and Image J analysis software was used for this study. The calibration curve of GafChromic EBT3 film was generated with known dose values for 14 mm collimator helmet using ImageJ software. The collimator helmet factor (CHF) for 4mm, 8mm and 14 mm collimator helmets were measured by normalizing dose rates of 4mm, 8mm and 14 mm to the dose rate of 18 mm collimator helmet using the previously generated calibration curve. The measured CHF was compared to Elekta reference value and previously published mean values. RESULTS: The measured CHFs were 0.896, 0.958, and 0.986 for 4mm, 8mm and 14mm collimators respectively. The percentage difference obtained was 1.7 %, 0.21 %, 0.1 % between measured values and reference values. CONCLUSION: The measurement of CHFs in LGK 4C unit using GafChromic EBT3 film and ImageJ software is a reliable method to verify the manufacturer quoted CHFs in routine quality assurance procedures.

Silver nanoparticle-base lateral flow immunoassay for rapid detection of Staphylococcal enterotoxin B in milk and honey.

A silver nanoparticle (AgNP)-based sandwich-type lateral flow immunoassay (LFIA) was evaluated for rapid detection of Staphylococcal enterotoxin B (SEB) in milk and honey. The role of trisodium citrate dihydrate (TSC) in the formation of Ag/TSC nanoparticles was established using UV-Vis spectroscopy. The association of silver with TSC in Ag/TSC nanoparticles was studied by the decrease in the intensity of anodic peak potential at 0.47 V and shift to 0.30 V in cyclic voltammetry (CV). The morphological, compositional and interaction studies of the AgNPs conjugated with the anti-SEB polyclonal antibody (Ag-sAb) was established using transmission electron microscopy (TEM) and X-ray photo electron spectroscopy (XPS) measurements. The visible detection limit and optical detection limit of the SEB test strip were 0.5 and 0.125 ppm, respectively, in SEB standard solution. This assay showed no cross-reaction with Staphylococcal enterotoxin A, Staphylococcal enterotoxin C or Salmonella typhi. Finally, the SEB test strip was effectively applied for the detection of SEB in spiked liquid milk and viscous honey, with optical detection limits of 0.25 and 0.5 ppm, respectively.

Monosynaptic inputs to specific cell types of the intermediate and deep layers of the superior colliculus.

The intermediate and deep layers of the midbrain superior colliculus (SC) are a key locus for several critical functions, including spatial attention, multisensory integration, and behavioral responses. While the SC is known to integrate input from a variety of brain regions, progress in understanding how these inputs contribute to SC-dependent functions has been hindered by the paucity of data on innervation patterns to specific types of SC neurons. Here, we use G-deleted rabies virus-mediated monosynaptic tracing to identify inputs to excitatory and inhibitory neurons of the intermediate and deep SC. We observed stronger and more numerous projections to excitatory than inhibitory SC neurons. However, a subpopulation of excitatory neurons thought to mediate behavioral output received weaker inputs, from far fewer brain regions, than the overall population of excitatory neurons. Additionally, extrinsic inputs tended to target rostral excitatory and inhibitory SC neurons more strongly than their caudal counterparts, and commissural SC neurons tended to project to similar rostrocaudal positions in the other SC. Our findings support the view that active intrinsic processes are critical to SC-dependent functions, and will enable the examination of how specific inputs contribute to these functions.

TMIS: a new image-based software application for the measurement of tear meniscus height.

PURPOSE: To present a new automated image recognition software for the measurement of tear meniscus height (TMH) and investigate its correlation and efficacy compared with an open-source software (NIH ImageJ) and manual evaluation. METHODS: A total of 520 slit lamp photographs, among which 276 were in ×16 magnification and 244 were ×40 magnified, captured from 138 eyes of 69 healthy subjects were assessed for TMH by the new automated Tear Meniscus Identification Software (TMIS), ImageJ and human graders. Images processing of TMIS included filtration, recognition and measurement of slit lamp photographs under certain algorithm, which output two measurement patterns, TMISM ax and TMISM ean . TMH measured by ImageJ software, considered as the reference value, was conducted by a masked observer while four masked ophthalmologists performed the manual evaluation. RESULTS: In both magnifications, TMH measured by TMISM ean showed similar values with ImageJ while manual evaluation demonstrated underestimated results, and a strong correlation was detected between TMIS and ImageJ. In ×16 magnified photographs, manually obtained TMH revealed a higher correlation with ImageJ, whereas a notably stronger correlation of TMIS with ImageJ was observed in ×40 photographs. Correspondingly, the accuracy for both TMISM ax and TMISM ean appeared to be lower than most doctors in ×16 slit lamp images, in contrast to a better precision of TMISM ean in ×40 ones. CONCLUSION: The new software displayed high accuracy and efficacy in ×40 magnification and TMISM ean pattern, suggesting the possibility of this automated TMH measurement platform to be a valid tool in dry eye screening and follow-up practice.

Temporomandibular disorders and mandibular vertical asymmetry.

Objective To analyze the association between mandibular vertical asymmetry and the presence of TMD in adult patients. Methods This case-control study recruited patients from the orthodontic clinic at FEBUAP. Patients were classified on the basis of diagnostic criteria for temporomandibular disorders (DC/TMD). Panoramic radiographs were measured to determine mandibular vertical asymmetry indices according to criteria described by Kjellberg and Habets. Both methods (DC/TMD and index criteria) were standardized a priori. Results A total of 56 patients were analyzed, of whom 25 (44.6%) met the inclusion criteria and were divided into two groups (TMD group, n = 15; non-TMD group, n = 10). The highest asymmetry index was observed in the condylar neck, followed by the total condylar asymmetry index, but without significant differences between groups (p > .05). Conclusion There is no association (p > .05) between temporomandibular disorders and mandibular vertical asymmetry.

ImageJ: A Free, Easy, and Reliable Method to Measure Leg Ulcers Using Digital Pictures.

Wound measurement to document the healing course of chronic leg ulcers has an important role in the management of these patients. Digital cameras in smartphones are readily available and easy to use, and taking pictures of wounds is becoming a routine in specialized departments. Analyzing digital pictures with appropriate software provides clinicians a quick, clean, and easy-to-use tool for measuring wound area. A set of 25 digital pictures of plain foot and leg ulcers was the basis of this study. Photographs were taken placing a ruler next to the wound in parallel with the healthy skin with the iPhone 6S (Apple Inc, Cupertino, CA), which has a camera of 12 megapixels using the flash. The digital photographs were visualized with ImageJ 1.45s freeware (National Institutes of Health, Rockville, MD; http://imagej.net/ImageJ ). Wound area measurement was carried out by 4 raters: head of the department, wound care nurse, physician, and medical student. We assessed intra- and interrater reliability using the interclass correlation coefficient. To determine intraobserver reliability, 2 of the raters repeated the measurement of the set 1 week after the first reading. The interrater model displayed an interclass correlation coefficient of 0.99 with 95% confidence interval of 0.999 to 1.000, showing excellent reliability. The intrarater model of both examiners showed excellent reliability. In conclusion, analyzing digital images of leg ulcers with ImageJ estimates wound area with excellent reliability. This method provides a free, rapid, and accurate way to measure wounds and could routinely be used to document wound healing in daily clinical practice.