Dose equivalent consideration from neutron contamination in modified radiotherapy vault: a Monte Carlo study.

Laminated barriers incorporating metal sheets provide effective protection for space-restricted radiotherapy centers. This study aimed to assess photoneutron contamination in smaller vaults protected by different compositions of multilayer barriers during simulated pelvic radiotherapy with 18 MV photon beams. Monte Carlo Simulations of 18 MV LINAC (Varian 2100 C/D) and Medical Internal Radiation Dose (MIRD) phantom were used to assess photoneutron contamination within reconstructed vaults incorporating different combinations of metal sheet and borated polyethylene (BPE) during pelvic radiotherapy. The findings highlight a 3.27 and 2.91 times increase in ambient neutron doseHn*(10) along the maze of reconstructed vaults that use lead and steel sheets, respectively, compared to concrete. TheHn*(10) outside the treatment room increased after incorporating a metal sheet, but it remained within the permissible limit of 20µSv/week for uncontrolled areas adjacent to the LINAC bunker, even with a workload of 1000Gy/week. Neutron equivalent doses in the patient's organs ranged from 0.22 to 0.96 mSv Gy-1. There is no notable distinction in the organ's neutron equivalent dose, fatal cancer risk, secondary radiation-induced cancer risk, and cancer mortality for various laminated barrier compositions. Furthermore, the use of metal sheets for vault wall reconstruction keeps the variation in cancer risk induced by photoneutrons below 6%, while risks of fatal cancer and cancer mortality vary less than 11%. While the metal portion of the laminated barrier raises the neutron dose, the addition of a BPE plate reduces concerns of increased effective dose and secondary malignancy risk.

A novel ternary magnetic nanobiocomposite based on tragacanth-silk fibroin hydrogel for hyperthermia and biological properties.

This study involves the development of a new nanocomposite material for use in biological applications. The nanocomposite was based on tragacanth hydrogel (TG), which was formed through cross-linking of Ca2+ ions with TG polymer chains. The utilization of TG hydrogel and silk fibroin as natural compounds has enhanced the biocompatibility, biodegradability, adhesion, and cell growth properties of the nanobiocomposite. This advancement makes the nanobiocomposite suitable for various biological applications, including drug delivery, wound healing, and tissue engineering. Additionally, Fe3O4 magnetic nanoparticles were synthesized in situ within the nanocomposite to enhance its hyperthermia efficiency. The presence of hydrophilic groups in all components of the nanobiocomposite allowed for good dispersion in water, which is an important factor in increasing the effectiveness of hyperthermia cancer therapy. Hemolysis and 3-(4,5 dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assays were conducted to evaluate the safety and efficacy of the nanobiocomposite for in-vivo applications. Results showed that even at high concentrations, the nanobiocomposite had minimal hemolytic effects. Finally, the hyperthermia application of the hybrid scaffold was evaluated, with a maximum SAR value of 41.2 W/g measured in the first interval.

Beam quality and the mystery behind the lower percentage depth dose in grid radiation therapy.

Grid therapy recently has been picking momentum due to favorable outcomes in bulky tumors. This is being termed as Spatially Fractionated Radiation Therapy (SFRT) and lattice therapy. SFRT can be performed with specially designed blocks made with brass or cerrobend with repeated holes or using multi-leaf collimators where dosimetry is uncertain. The dosimetric challenge in grid therapy is the mystery behind the lower percentage depth dose (PDD) in grid fields. The knowledge about the beam quality, indexed by TPR20/10 (Tissue Phantom Ratio), is also necessary for absolute dosimetry of grid fields. Since the grid may change the quality of the primary photons, a new [Formula: see text] should be evaluated for absolute dosimetry of grid fields. A Monte Carlo (MC) approach is provided to resolving the dosimetric issues. Using 6 MV beam from a linear accelerator, MC simulation was performed using MCNPX code. Additionally, a commercial grid therapy device was used to simulate the grid fields. Beam parameters were validated with MC model for output factor, depth of maximum dose, PDDs, dose profiles, and TPR20/10. The electron and photon spectra were also compared between open and grid fields. The dmax is the same for open and grid fields. The PDD with grid is lower (~ 10%) than the open field. The difference in TPR20/10 of open and grid fields is observable (~ 5%). Accordingly, TPR20/10 is still a good index for the beam quality in grid fields and consequently choose the correct [Formula: see text] in measurements. The output factors for grid fields are 0.2 lower compared to open fields. The lower depth dose with grid therapy is due to lower depth fluence with scatter radiation but it does not impact the dosimetry as the calibration parameters are insensitive to the effective beam energies. Thus, standard dosimetry in open beam based on international protocol could be used.

Experimental investigation of the effect of landfill leachate on the mechanical and cracking behavior of polypropylene fiber-reinforced compacted clay liner.

This paper aims to investigate the effect of leachate on the geotechnical parameters and the cracking behavior of compacted clay liners (CCLs) containing different percentages of polypropylene fibers. Accordingly, 200 compacted clay samples were reinforced with different percentages of fiber contents (FC) (i.e., 0, 0.5, 0.75, and 1%) and prepared with water or leachate to conduct different laboratory tests. First, the physical properties of the clay were determined. Then, the shear strength parameters (i.e., cohesion and friction angle), unconfined compressive strength, and the hydraulic permeability were determined subjected to water or leachate. Notably, the cracking behavior was modeled using visual images of the samples. The leachate increased desiccation cracks in the natural soil from 0.425 to nearly 1.111%. However, the addition of 0.5% (in the case of water) and 1% (in the case of leachate) fiber to the soil reduced the surface desiccation cracks in clay liners to about 0.185 and 0.352%, respectively. In both water- or leachate-prepared samples, the addition of fibers significantly increased the cohesion and friction angle. The shear strengths of the unreinforced leachate-prepared samples were lower than those of the water-prepared samples. The shear strength and unconfined compressive strength of all specimens increased with increasing fiber percentage. The presence of fibers in all samples caused more ductile behavior. The required amount of energy to achieve the maximum strength in the samples increased with increasing FC. By increasing the percentage of fibers, the permeability of the natural soil and the leachate-prepared samples increased. However, the highest permeability was observed in the leachate-prepared samples containing 1% fibers of 8.3 × 10-10 m/s, which is less than 10-9 m/s (maximum allowable permeability for clay liners). Finally, the obtained results were satisfactorily confirmed by scanning electron microscopy (SEM) analysis.

Dual application of Polyvinyl Alcohol Glutaraldehyde Methylthymol Blue Fricke hydrogel in clinical practice: Surface dosimeter and bolus.

An essential issue is an accurate evaluation of surface dose distribution for such sensitive treatments. This work aimed to feasibility of the dual application of the Ferrous Polyvinyl Alcohol Glutaraldehyde Methylthymol Blue (PVA-GTA-MTB) gel as a bolus compensator and surface dosimeter in breast radiotherapy. The differences between the surface dose measured using PVA-GTA-MTB gel and film dosimetry in the medial and lateral parts of the breast were 3.74% and 4.18%, respectively. A qualitative comparison of the isodose curves showed that the PVA-GTA-MTB bolus creates a uniform dose distribution similar to the superflab bolus in the target volume.

Network-Based and Machine-Learning Approaches Identify Diagnostic and Prognostic Models for EMT-Type Gastric Tumors.

The microsatellite stable/epithelial-mesenchymal transition (MSS/EMT) subtype of gastric cancer represents a highly aggressive class of tumors associated with low rates of survival and considerably high probabilities of recurrence. In the era of precision medicine, the accurate and prompt diagnosis of tumors of this subtype is of vital importance. In this study, we used Weighted Gene Co-expression Network Analysis (WGCNA) to identify a differentially expressed co-expression module of mRNAs in EMT-type gastric tumors. Using network analysis and linear discriminant analysis, we identified mRNA motifs and microRNA-based models with strong prognostic and diagnostic relevance: three models comprised of (i) the microRNAs miR-199a-5p and miR-141-3p, (ii) EVC/EVC2/GLI3, and (iii) PDE2A/GUCY1A1/GUCY1B1 gene expression profiles distinguish EMT-type tumors from other gastric tumors with high accuracy (Area Under the Receiver Operating Characteristic Curve (AUC) = 0.995, AUC = 0.9742, and AUC = 0.9717; respectively). Additionally, the DMD/ITGA1/CAV1 motif was identified as the top motif with consistent relevance to prognosis (hazard ratio > 3). Molecular functions of the members of the identified models highlight the central roles of MAPK, Hh, and cGMP/cAMP signaling in the pathology of the EMT subtype of gastric cancer and underscore their potential utility in precision therapeutic approaches.

How much should you worry about contaminant neutrons in spatially fractionated grid radiation therapy?

Neutron contamination in radiation therapy is of concern in treatment with high-energy photons (> 10 MV). With the development of new radiotherapy modalities such as spatially fractionated grid radiation therapy (SFGRT) or briefly grid radiotherapy, more studies are required to evaluate the risks associated with neutron contamination. In 15 MV SFGRT, high-Z materials such as lead and cerrobend are used as the block on the tray of linear accelerator (linac) which can probably increase the photoneutron production. On the other hand, the high-dose fractions (10-20 Gy) used in SFGRT can induce high neutron contamination. The current study was devoted to addressing these concerns via compression of neutron fluence (Φn) and ambient dose equivalent ([Formula: see text]) at the patient table and inside the maze between SFGRT and conventional fractionated radiation therapy (CFRT). The main components of the 15 MV Siemens Primus equipped with different grids and located inside a typical radiotherapy bunker were simulated by the MCNPX® Monte Carlo code. Evidence showed that the material used for grid construction does not significantly increase neutron contamination inside the maze. However, at the end of the maze, neutron contamination in SFGRT is significantly higher than in CFRT. In this regard, a delay time of 15 minutes after SFGRT is recommended for all radiotherapy staff before entering the maze. It can be also concluded that [Formula: see text] at the patient table is at least 10 times more pronounced than inside the maze. Therefore, the patient is more at risk of neutrons compared to the staff. The [Formula: see text] at the isocenter in SFGRT with grids made of lead and cerrobend was nearly equal to CFRT. Nevertheless, it was dramatically lower than in CFRT by 30% if the brass grid is used. Accordingly, SFGRT with the brass grid is recommended, from radiation protection aspects.

Semi-experimental assessment of neutron equivalent dose and secondary cancer risk for off-field organs in glioma patients undergoing 18-MV radiotherapy.

Neutron contamination as a source of out-of-field dose in radiotherapy is still of concern. High-energy treatment photons have the potential to overcome the binding energy of neutrons inside the nuclei. Fast neutrons emitting from the accelerator head can directly reach the patient's bed. Considering that modern radiotherapy techniques can increase patient survival, concerns about unwanted doses and the lifetime risk of fatal cancer remain strong or even more prominent, especially in young adult patients. The current study addressed these concerns by quantifying the dose and risk of fatal cancer due to photo-neutrons for glioma patients undergoing 18-MV radiotherapy. In this study, an NRD model rem-meter detector was used to measure neutron ambient dose equivalent, H*(10), at the patient table. Then, the neutron equivalent dose received by each organ was estimated concerning the depth of each organ and by applying depth dose corrections to the measured H*(10). Finally, the effective dose and risk of secondary cancer were determined using NCRP 116 coefficients. Evidence revealed that among all organs, the breast (0.62 mSv/Gy) and gonads (0.58 mSv/Gy) are at risk of photoneutrons more than the other organs in such treatments. The neutron effective dose in the 18-MV conventional radiotherapy of the brain was 13.36 mSv. Among all organs, gonads (6.96 mSv), thyroid (1.86 mSv), and breasts (1.86 mSv) had more contribution to the effective dose, respectively. The total secondary cancer risk was estimated as 281.4 cases (per 1 million persons). The highest risk was related to the breast and gonads with 74.4 and, 34.8 cases per 1 million persons, respectively. Therefore, it is recommended that to prevent late complications (secondary cancer and genetic effects), these organs should be shielded from photoneutrons. This procedure not only improves the quality of the patient's personal life but also the healthy childbearing in the community.

Measurement of neutron equivalent dose in the thyroid, chiasma, and lens for patients undergoing pelvic radiotherapy: A phantom study.

Radiotherapy is one of the most common methods for treating malignant diseases, whose ultimate goal is to deliver lethal doses to tumor cells. One of the unwanted consequences of radiotherapy is secondary radiation outside the treatment field, which imposes additional doses to healthy tissues and organs, specifically neutron doses, which we aim to evaluate. Therefore, this study aims to measure the fast neutron equivalent dose and the risk of secondary cancer in the thyroid, chiasm, and lenses in the treatment of the pelvic area. In this study, CR-39 detectors were used to measure the equivalent fast neutron dose in different sections of the RANDO Phantom (thyroid, chiasm, and lenses) and were irradiated by the energy of 18 MV on Varian Clinac 2100 C-D linear accelerator. CR-39 detectors were calibrated with predetermined doses by an Am-Be neutron source. Then, after etching and reading processes, the equivalent dose of fast neutrons was determined. According to the results, the fast neutron doses in the thyroid, right and left eye lenses, and chiasm were 0.613 ± 0.024, 0.835 ± 0.040, 0.866 ± 0.016, and 0.685 ± 0.045 mSv/Gy, respectively. Moreover, the secondary cancer risks in the unshielded organs are 0.004, 0.029, 0.030, and 0.025 for the thyroid, right and left eye lenses, and chiasm, respectively. In conclusion, the contribution of neutrons to the secondary doses in the out-of-field organs is significant and should not be ignored.

Prospects and challenges of cancer systems medicine: from genes to disease networks.

It is becoming evident that holistic perspectives toward cancer are crucial in deciphering the overwhelming complexity of tumors. Single-layer analysis of genome-wide data has greatly contributed to our understanding of cellular systems and their perturbations. However, fundamental gaps in our knowledge persist and hamper the design of effective interventions. It is becoming more apparent than ever, that cancer should not only be viewed as a disease of the genome but as a disease of the cellular system. Integrative multilayer approaches are emerging as vigorous assets in our endeavors to achieve systemic views on cancer biology. Herein, we provide a comprehensive review of the approaches, methods and technologies that can serve to achieve systemic perspectives of cancer. We start with genome-wide single-layer approaches of omics analyses of cellular systems and move on to multilayer integrative approaches in which in-depth descriptions of proteogenomics and network-based data analysis are provided. Proteogenomics is a remarkable example of how the integration of multiple levels of information can reduce our blind spots and increase the accuracy and reliability of our interpretations and network-based data analysis is a major approach for data interpretation and a robust scaffold for data integration and modeling. Overall, this review aims to increase cross-field awareness of the approaches and challenges regarding the omics-based study of cancer and to facilitate the necessary shift toward holistic approaches.

Feasibility of 18-MV grid therapy from radiation protection aspects: unwanted dose and fatal cancer risk caused by photoneutrons and scattered photons.

PURPOSE: Photoneutron production is a common concern when using 18-MV photon beams in radiation therapy. In Spatially Fractionated Grid Radiation Therapy (SFGRT), the grid block in the collimation system modifies the neutron production, photon scattering, and electron contamination in and out of the radiation field. Such an effect was studied with grids made of different high-Z materials by Monte Carlo simulations. The results were also used to evaluate the lifetime risk of fatal cancers. METHODS: MCNPX® code (2.7.0 extensions) was employed to simulate an 18-MV LINAC (Varian 2100 C/D). Three types of grid made of brass, cerrobend, and lead were used to study the neutron and electron fluence. Output factors for each grid with different field sizes were calculated. A revised female MIRD phantom with an 8-cm spherical tumor inside the liver was used to estimate the dose to the tumor and the critical organs. A 20-Gy SFGRT plan with Anterior Posterior (AP) - Posterior Anterior (PA) grid beams was compared with a Conventional Fractionated Radiation Therapy (CFRT) plan which delivered 40-Gy to the tumor by AP-PA open beams. Neutron equivalent dose, photon equivalent dose, as well as lifetime risks of fatal cancer were calculated in the organs at risk. RESULTS: The grid blocks reduced the fluence of contaminant electrons inside the treatment field by more than 50%. The neutron fluences per electron-history in SFGRT plans with brass, cerrobend and lead were on average 55%, 31% and 31% less than that of the CFRT plan, respectively. However, when converting to fluences per delivered dose (Gy), the cerrobend and lead grid may incur higher neutron dose for 20 × 20 cm2 field size and above. The changes in neutron mean energy, as well as the correlated radiation weighting factors, were insignificant. The total risk due to the photoneutrons in the SFGRT plans was 87% or lower than that in the CFRT plans. In both SFGRT and CFRT plans, the contribution of the primary and scattered photons to the fatal cancer risk was 2 times or more than the photoneutrons. The total risks from photons in SFGRT with brass, cerrobend, and lead blocks were 1.733, 1.374, and 1.260%, respectively, which were less than 30% of the total photon-risk in CFRT (5.827%). CONCLUSION: In the brass, cerrobend, and lead grids, the attenuation of photoneutrons outweighs its photoneutron production in 18-MV SFGRT. The total cancer risks from photons and photoneutrons in the SFGRT plans were 30% or less of the risks in the CFRT plans (5.911%). Using 18 MV photon beams with brass, cerrobend, and lead grid blocks is still a feasible option for SFGRT.

Monte Carlo evaluation of out-of-field dose in 18 MV pelvic radiotherapy using a simplified female MIRD phantom.

This study was devoted to determining the unwanted dose due to scattered photons to the out-of-field organs and subsequently estimate the risk of secondary cancers in the patients undergoing pelvic radiotherapy. A typical 18 MV Medical Linear Accelerator (Varian Clinac 2100 C/D) was modeled using MCNPX®code to simulate pelvic radiotherapy with four treatment fields: anterior-posterior, posterior-anterior, right lateral, left lateral. Dose evaluation was performed inside Medical Internal Radiation Dose (MIRD) revised female phantom. The average photon equivalent dose in out-of-field organs is 8.53 mSv Gy-1, ranging from 0.17 to 72.11 mSv Gy-1, respectively, for the organs far from the Planning Treatment Volume (Brain) and those close to the treatment field (Colon). Evidence showed that colon with 4.3049% and thyroid with 0.0020% have the highest and lowest risk of secondary cancer, respectively. Accordingly, this study introduced the colon as an organ with a high risk of secondary cancer which should be paid more attention in the follow-up of patients undergoing pelvic radiotherapy. The authors believe that this simple Monte Carlo (MC) model can be also used in other radiotherapy plans and mathematical phantoms with different ages (from childhood to adults) to estimate the out-of-field dose. The extractable information by this simple MC model can be also employed for providing libraries for user-friendly applications (e.g. '.apk') which in turn increase the public knowledge about fatal cancer risk after radiotherapy and subsequently decrease the concerns in this regard among the public.

A Homozygous Truncating Mutation in NALCN Causing IHPRF1: Detailed Clinical Manifestations and a Review of Literature.

Infantile hypotonia, with psychomotor retardation and characteristic facies 1 (IHPRF1), is a rare disorder characterized by global developmental delay and dysmorphic features. This syndrome is caused by genetic anomalies within the NALCN gene. The current report examines a 9-year-old female IHPRF1 patient. Our objective was to contribute to the delineation of the underlying factors influencing this rare condition. Whole exome sequencing (WES) was utilized to identify the disease-causing mutation in the affected individual. Subsequently, Sanger sequencing was performed for the patient, her parents, and two close relatives in order to confirm the detected mutation. Moreover, detailed clinical examinations including EEG, echocardiography, and biochemical/physical tests were carried out to elucidate the effects of the mutation. WES identified a homozygous nonsense mutation in the NALCN gene (c.2563C>T p.R855X). This mutation was confirmed by Sanger sequencing in the patient and her family members and segregated with the autosomal recessive inheritance pattern of IHPRF1. Moreover, genotype-phenotype correlation analysis confirmed the disease-causing nature of this mutation. The current report provides the first detailed description of a patient with this homozygous nonsense mutation (c.2563C>T p.R855X) and expands the clinical spectrum of IHPRF1 disease. Possible influences of sex and other factors on this disease are discussed and a review of the literature is also provided.

Grid therapy vs. conventional radiotherapy - 18 MV treatments: Photoneutron contamination along the maze of a linac bunker.

The present Monte Carlo study was devoted to the comparison of photoneutron contamination (per 1 Gy photon dose), along the maze of a radiotherapy bunker, between two 18-MV modalities: grid therapy (with grids made of brass, cerrobend, and lead) and conventional radiotherapy. It was turned out that both in grid therapy and in conventional radiotherapy, with increasing distance from the entrance of treatment hall (toward the maze entrance), fluence and ambient dose equivalent of neutrons decrease. Evidence also shows that in grid therapy, independent of materials used in the grid construction, photoneutron contamination along the maze is 45±6 % larger than conventional radiotherapy.

Essential considerations for accurate evaluation of photoneutron contamination in Radiotherapy.

Nowadays, high-energy X-rays produced by medical linear accelerators (LINACs) are widely used in many Radiation Therapy (RT) centers. High-energy photons (> 8 MeV) produce undesired neutrons in the LINAC head which raise concerns about unwanted neutron dose to the patients and RT personnel. Regarding the significance of radiation protection in RT, it is important to evaluate photoneutron contamination inside the RT room. Unfortunately, neutron dosimeters used for this purpose have limitations that can under the best conditions cause to > 10% uncertainty. In addition to this uncertainty, the present Monte Carlo (MC) study introduces another uncertainty in measurements (nearly up to 20%) when neutron ambient dose equivalent (Hn*(10)) is measured at the patient table or inside the maze and the change in neutron energy is ignored. This type of uncertainty can even reach 35% if Hn*(10) is measured by dosimeters covered by a layer of 10B as converter. So, in these cases, neglecting the change in neutron energy can threaten the credibility of measured data and one should attend to this energy change in order to reduce measurement uncertainty to the possible minimum. This study also discusses the change in neutron spectra and Hn*(10) at the patient table caused by removing a typical RT room from MC simulations. Under such conditions, neutron mean energy (En) overestimated by 0.2-0.4 MeV at the patient table. Neutron fluence (φn) at the isocenter (IC) was underestimated by 23-54% for different field sizes that caused Hn*(10) to be miscalculated up to 24%. This finding informs researchers that for accurate evaluation of Hn*(10) at the patient table, simulating the RT room is an effective parameter in MC studies.

Photoneutron Dose Estimation in GRID Therapy Using an Anthropomorphic Phantom: A Monte Carlo Study.

BACKGROUND: In the past, GRID therapy was used as a treatment modality for the treatment of bulky and deeply seated tumors with orthovoltage beams. Now and with the introduction of megavoltage beams to radiotherapy, some of the radiotherapy institutes use GRID therapy with megavoltage photons for the palliative treatment of bulky tumors. Since GRID can be a barrier for weakening the photoneutrons produced in the head of medical linear accelerators (LINAC), as well as a secondary source for producing photoneutrons, therefore, in terms of radiation protection, it is important to evaluate the GRID effect on photoneutron dose to the patients. METHODS: In this study, using the Monte Carlo code MCNPX, a full model of a LINAC was simulated and verified. The neutron source strength of the LINAC (Q), the distributions of flux (φ), and ambient dose equivalent (H*[10]) of neutrons were calculated on the treatment table in both cases of with/without the GRID. Finally, absorbed dose and dose equivalent of neutrons in some of the tissues/organs of MIRD phantom were computed with/without the GRID. RESULTS: Our results indicate that the GRID increases the production of the photoneutrons in the LINAC head only by 0.3%. The calculations in the MIRD phantom show that neutron dose in the organs/tissues covered by the GRID is on average by 48% lower than conventional radiotherapy. In addition, in the uncovered organs (by the GRID), this amount is reduced to 25%. CONCLUSION: Based on the findings of this study, in GRID therapy technique compared to conventional radiotherapy, the neutron dose in the tissues/organs of the body is dramatically reduced. Therefore, there will be no concern about the GRID effect on the increase of unwanted neutron dose, and consequently the risk of secondary cancer.

Automated detection and cell density assessment of keratocytes in the human corneal stroma from ultrahigh resolution optical coherence tomograms.

Keratocytes are fibroblast-like cells that maintain the optical clarity and the overall health of the cornea. The ability to measure precisely their density and spatial distribution in the cornea is important for the understanding of corneal healing processes and the diagnostics of some corneal disorders. A novel computerized approach to detection and counting of keratocyte cells from ultra high resolution optical coherence tomography (UHR-OCT) images of the human corneal stroma is presented. The corneal OCT data is first processed using a state-of-the-art despeckling algorithm to reduce the effect of speckle on detection accuracy. A thresholding strategy is then employed to allow for improved delineation of keratocyte cells by suppressing similarly shaped features in the data, followed by a second-order moment analysis to identify potential cell nuclei candidates. Finally, a local extrema strategy is used to refine the candidates to determine the locations and the number of keratocyte cells. Cell density distribution analysis was carried in 3D UHR-OCT images of the human corneal stroma, acquired in-vivo. The cell density results obtained using the proposed novel approach correlate well with previous work on computerized keratocyte cell counting from confocal microscopy images of human cornea.