Estimation Of Cell Number Research Articles

RNA Polymerase II hypertranscription in cancer FFPE samples.

Hypertranscription is common in human cancers and predicts poor prognosis. However detection of hypertranscription is indirect, relying on accurately quantifying mRNA levels and estimating cell numbers. Previously, we introduced FFPE-CUTAC, a genome-wide method for mapping RNA Polymerase II (RNAPII) in formalin-fixed paraffin-embedded (FFPE) sections. Here we use FFPE-CUTAC to demonstrate genome-wide hypertranscription both in transgene-driven mouse gliomas and in assorted human tumors at active regulatory elements and replication-coupled histone genes with reduced mitochondrial DNA abundance. FFPE-CUTAC identified RNAPII-bound regulatory elements shared among diverse cancers and readily categorized human tumors despite using very small samples and low sequencing depths. Remarkably, RNAPII FFPE-CUTAC identified de novo and precisely mapped HER2 amplifications punctuated by likely selective sweeps including genes encoding direct positive regulators of RNAPII itself. Our results demonstrate that FFPE-CUTAC measurements of hypertranscription and classifications of tumors using small sections provides an affordable and sensitive genome-wide strategy for personalized medicine.

Gene-dosage- and sex-dependent differences in the prodromal-Like phase of the F344tgHD rat model for Huntington disease.

In Huntington disease (HD) the prodromal phase has been increasingly investigated and is currently in focus for early interventional treatments. Also, the influence of sex on disease progression and severity in patients is under discussion, as a sex-specific impact has been reported in transgenic rodent models for HD. To this end, we have been studying these aspects in Sprague Dawley rats transgenic for HD. Here, we took up on the congenic F344tgHD rat model, expressing a fragmented Htt construct with 51 CAG repeats on an inbred F344 rat background and characterized potential sexual dimorphism and gene-dosage effects in rats during the pre-symptomatic phase (1-8 months of age). Our study comprises a longitudinal phenotyping of motor function, emotion and sensorimotor gating, as well as screening of metabolic parameters with classical and automated assays in combination with investigation of molecular HD hallmarks (striatal cell number and volume estimation, appearance of HTT aggregates). Differences between sexes became apparent during middle age, particularly in the motor and sensorimotor domains. Female individuals were generally more active, demonstrated different gait characteristics than males and less anxiolytic-like behavior. Alterations in both the time course and affected behavioral domains varied between male and female F344tgHD rats. First subtle behavioral anomalies were detected in transgenic F344tgHD rats prior to striatal MSN cell loss, revealing a prodromal-like phase in this model. Our findings demonstrate that the congenic F344tgHD rat model shows high face-validity, closely resembling the human disease's temporal progression, while having a relatively low number of CAG repeats, a slowly progressing pathology with a prodromal-like phase and a comparatively subtle phenotype. By differentiating the sexes regarding HD-related changes and characterizing the prodromal-like phase in this model, these findings provide a foundation for future treatment studies.

HPLC-Based Detection of Two Distinct Red Tide Causative Species (Mesodinium rubrum and Margalefidinium polykrikoides) in the South Sea of Korea

Various approaches have been applied to red tide monitoring in Korea since reliable information on phytoplankton communities is crucial. In this study, we employed a high-performance liquid chromatography (HPLC) method to analyze two types of red tide, Mesodinium rubrum and Margalefidinium polykrikoides (also known as Cochlodinium polykrikoides), along the southern coasts of Korea. During the M. rubrum red tide on 8 August 2022, an unusual dominance of cryptophytes was observed, being the most dominant phytoplankton group. A significant positive correlation was found between alloxanthin concentrations, a marker pigment of cryptophytes, and M. rubrum cell numbers (p < 0.01, r = 0.830), indicating that HPLC-derived alloxanthin concentrations can serve as a valuable indicator for identifying red tides caused by M. rubrum and estimating cell numbers. However, it is crucial to consider the temporal dynamics of the prey–predator relationship between cryptophytes and M. rubrum. Further investigation is required to understand the environmental conditions that promote cryptophyte predominance and their role in M. rubrum red tide development. In the second field campaign on 29 August 2022, we observed a significant correlation between the concentration of peridinin, a marker pigment for dinoflagellates, and M. polykrikoides cell numbers (p < 0.01, r = 0.663), suggesting that peridinin can serve as a reliable indicator of M. polykrikoides red tides. In conclusion, HPLC-derived pigments, namely alloxanthin and peridinin, can be used to effectively monitor red tides caused by M. rubrum and M. polykrikoides, respectively. However, to overcome certain methodological limitations of HPLC, future studies should explore additional markers or analytical techniques capable of differentiating M. polykrikoides from other coexisting dinoflagellate species. Furthermore, the broad applicability of our method requires thorough investigation in diverse ecosystems to fully comprehend its scope and limitations. Future research should focus on evaluating the method’s efficacy in different contexts, accounting for the distinct traits of the ecosystems under consideration.

Novel stereological method for estimation of cell counts in 3D collagen scaffolds

Current methods for assessing cell proliferation in 3D scaffolds rely on changes in metabolic activity or total DNA, however, direct quantification of cell number in 3D scaffolds remains a challenge. To address this issue, we developed an unbiased stereology approach that uses systematic-random sampling and thin focal-plane optical sectioning of the scaffolds followed by estimation of total cell number (StereoCount). This approach was validated against an indirect method for measuring the total DNA (DNA content); and the Bürker counting chamber, the current reference method for quantifying cell number. We assessed the total cell number for cell seeding density (cells per unit volume) across four values and compared the methods in terms of accuracy, ease-of-use and time demands. The accuracy of StereoCount markedly outperformed the DNA content for cases with ~ 10,000 and ~ 125,000 cells/scaffold. For cases with ~ 250,000 and ~ 375,000 cells/scaffold both StereoCount and DNA content showed lower accuracy than the Bürker but did not differ from each other. In terms of ease-of-use, there was a strong advantage for the StereoCount due to output in terms of absolute cell numbers along with the possibility for an overview of cell distribution and future use of automation for high throughput analysis. Taking together, the StereoCount method is an efficient approach for direct cell quantification in 3D collagen scaffolds. Its major benefit is that automated StereoCount could accelerate research using 3D scaffolds focused on drug discovery for a wide variety of human diseases.

Estimates of total neuron number show that neonatal ethanol causes immediate and lasting neuron loss in cortical and subcortical areas.

In neonatal brain development there is a period of normal apoptotic cell death that regulates adult neuron number. At approximately the same period, ethanol exposure can cause a dramatic spike in apoptotic cell death. While ethanol-induced apoptosis has been shown to reduce adult neuron number, questions remain about the regional selectivity of the ethanol effect, and whether the brain might have some capacity to overcome the initial neuron loss. The present study used stereological cell counting to compare cumulative neuron loss 8 h after postnatal day 7 (P7) ethanol treatment to that of animals left to mature to adulthood (P70). Across several brain regions we found that the reduction of total neuron number after 8 h was as large as that of adult animals. Comparison between regions revealed that some areas are more vulnerable, with neuron loss in the anterior thalamic nuclei > the medial septum/vertical diagonal band, dorsal subiculum, and dorsal lateral geniculate nucleus > the mammillary bodies and cingulate cortex > whole neocortex. In contrast to estimates of total neuron number, estimates of apoptotic cell number in Nissl-stained sections at 8 h after ethanol treatment provided a less reliable predictor of adult neuron loss. The findings show that ethanol-induced neonatal apoptosis often causes immediate neuron deficits that persist in adulthood, and furthermore suggests that the brain may have limited capacity to compensate for ethanol-induced neuron loss.

The Estimation of Blue-green algae Cell Count and Phycocyanin Concentration using Fluorescent Concentration analyzer

Objectives:The purpose of this study is to provide basic data for the development of a rapid and easy to measure alternative indicator for estimating the number of cyanobacteria cells in the field.Methods:Phycocyanin, algal cell number, and water quality items were measured at the left, middle, and right points of Maery Bridge near the Mulgeum and Maery algae monitoring network located downstream of the Nakdong River. The phycocyanin concentration(PC) was measured in the field using the LISST-HAB sensor, which is a fluorescence concentration analyzer, and comparative analysis was performed with the number of algal cells by algae species.Results and Discussion:As a limiting factor for blue-green algae occurrence, the correlation coefficient (r) between T-P and Blue-green algae (BGA) was 0.58 (≥0.3). Phycocyanin and Blue-green algae showed a high significant correlation at 0.96 (≥0.3). Prob (F-statistics) of the blue-green algae cell count estimation formula using phycocyanin was less than 0.05, showing a significant result. The estimation formula was estimated BGA(cells/mL) = 326.95×PC+50,484. As a result of the analysis of the predicted and measured values of Blue-green algae, r<sup>2</sup> was 0.988, indicating a high coefficient of determination and 92.5% accuracy.Conclusion: The correlation analysis with the number of cyanobacteria showed a high significant correlation with phycocyanin of 0.96 (≥0.3), followed by T-P, a limiting factor for the occurrence of cyanobacteria, of 0.58 (≥0.3) indicating a significant correlation. In addition, the formula for estimating the number of blue-green algae cells using phycocyanin was found to be BGA (cells/mL)=326.95×PC+50,484 in which r<sup>2</sup> was 0.927, and Prob (F-statistics) was less than 0.05. As a result of analyzing the predicted and measured values ​​of cyanobacteria cell numbers, r<sup>2</sup> was 0.988, indicating a high coefficient of determination, with accuracy of 92.5%. Through this results, it reveals that the number of cyanobacteria cells can be estimated using the sensor of the fluorescence concentration analyzer at the target point by substituting the phycocyanin concentration into the blue-green algae cell number estimation formula.

Uncertainties of cell number estimation in cyanobacterial colonies and the potential use of sphere packing

Cyanobacteria are notorious bloom formers causing various water quality concerns, such as toxin production, extreme diurnal variation of oxygen, pH, etc., therefore, their monitoring is essential to protect the ecological status of aquatic systems. Cyanobacterial cell counts and biovolumes are currently being used in water management and water quality alert systems. In this study, we investigated the accuracy of traditional colonial biovolume and cell count estimation approaches used in everyday practice. Using shape realistic 3D images of cyanobacterial colonies, we demonstrated that their shape cannot be approximated by ellipsoids. We also showed that despite the significant relationship between overall colony volume and cell biovolumes, because of the considerable scatter of cell count data the regressions give biased estimates for cyanobacterial cell counts. We proposed a novel approach to estimate cell counts in colonies that was based on the random close sphere packing method. This method provided good results only in those cases when overall colony volumes could be accurately measured. The visual investigation of colonies done by skilled experts has given precise but lower estimates for cell counts. The estimation results of several experts were surprisingly good, which suggests that this capability can be improved and estimation bias can be reduced to the level acceptable for water quality estimations.

The Effects of Graded Levels of Calorie Restriction: XVIII.Tissue-Specific Changes in Cell Size and Number in Response to Calorie Restriction.

Calorie restriction (CR) without malnutrition increases the health and life span of diverse taxa. The mechanism(s) behind CR are debated but may be directly linked to body composition changes that maintain energy balance. During a deficit, energy is primarily obtained from white adipose tissue (WAT; utilized) while other tissues remain unchanged (protected) or grow (invested) relative to body mass. The changes in mass of 6 tissues from 48 male C57BL/6 mice following 3-month graded (10%, 20%, 30%, or 40%) CR or fed ad libitum for 12 or 24 hours a day were related to cell size (hypo/hypertrophy) and/or number (hypo/hyperplasia). Tissues studied were retroperitoneal and subcutaneous WAT, brown adipose tissue (BAT; utilized), lungs (protected), and stomach and cecum (invested). Methodology was based on number of nuclei/tissue equaling the number of cells. Extracted DNA was quantified and used to estimate cell numbers (total DNA/DNA per diploid nucleus) and size (tissue mass/nuclei number). WAT utilization was caused solely by hypotrophy whereas BAT utilization resulted from reduced cell number and size. WAT cell size positively correlated with circulating hormones related to energy balance, and BAT cell number and size positively correlated with body temperature. No changes were found in the lungs, consistent with their protected status, whereas hyperplasia appeared to be the dominant mechanism for invested alimentary-tract tissues. These findings indicate the pattern of change of cell size and number across increasing levels of short-term CR is tissue-specific.

Posology and Serum-/Xeno-Free Engineered Adipose Stromal Cells Cell Sheets

Well-characterized adipose stem cells and chemically defined culture media are important factors that control the production of the cell sheet, used in translational medicine. In this study, we have developed and engineered multilayer adipose stem cell cell sheets (ASCCSs) using chemically defined/serum-free culture media: undifferentiated or differentiated into osteoblasts and chondrocytes. In addition, using the cell sheet transmittance, we estimated the number of cells per cell sheet. Undifferentiated ASCCSs were engineered in 10 days, using serum-free/xeno-free culture media. They were CD29+, CD73+, CD90+, CD105+, HLA-A+, and HLA-DR-. ASCCSs differentiated into chondrocytes and osteoblasts were also engineered using chemically defined and animal-free culture media, in only 14 days. The addition of an ROCK inhibitor improved the chondrocyte cell sheet engineering. The decrease in the cell sheet transmittance rate was higher for the osteoblast cell sheets due to the intracellular Ca2+ accumulation. The estimation of cell number per cell sheet was carried out with the transmittance, which will provide important information for cell sheet posology. In conclusion, three types of ASCCSs were engineered using serum-free, xeno-free culture media, expressing their specific markers. Their transmittance measurement allowed estimating the number of cells per cell sheet, with a non-invasive methodology.

3101 – HEMATOPOIESIS IN NUMBERS

Hematopoiesis is a dynamic process in which stem and progenitor cells give rise to the ∼10^13 blood and immune cells distributed throughout the human body. We argue that a quantitative description of hematopoiesis can help consolidate existing data, identify knowledge gaps, and generate new hypotheses. Here, we review known numbers in murine and, where possible, human hematopoiesis, and consolidate murine numbers into a set of reference values. We present estimates of cell numbers, division and differentiation rates, cell size, and macromolecular composition for each hematopoietic cell type. We also propose guidelines to improve the reporting of measurements and highlight areas in which quantitative data are lacking. Overall, we show how quantitative approaches can be used to understand key properties of hematopoiesis. Hematopoiesis is a dynamic process in which stem and progenitor cells give rise to the ∼10^13 blood and immune cells distributed throughout the human body. We argue that a quantitative description of hematopoiesis can help consolidate existing data, identify knowledge gaps, and generate new hypotheses. Here, we review known numbers in murine and, where possible, human hematopoiesis, and consolidate murine numbers into a set of reference values. We present estimates of cell numbers, division and differentiation rates, cell size, and macromolecular composition for each hematopoietic cell type. We also propose guidelines to improve the reporting of measurements and highlight areas in which quantitative data are lacking. Overall, we show how quantitative approaches can be used to understand key properties of hematopoiesis.

Rough estimation of cell numbers in 5G networks using simple mathematical calculations

This paper focuses on the problem of 5G network cell planning. In addition, it presents an example of a rough estimation of the required number of cells or base stations in a certain area for arbitrary number of users who are provided with a certain bandwidth per user within these cells. The cell number estimation is the initial step and the essence of planning and implementation of 5G network in an area. It is helpful for the operators to create and take into the account business plans in order to fully implement the network as a function of number of users which have to be served. Considering that, knowing the rough number of 5G base stations per user is very important for pre-sale activities and eventually necessity for widening of the initial investments. Therefore, the paper presents four scenarios that include different network parameters. Depending on the network parameters, the required number of base stations in a certain area changes. Given scenarios are examples of one 5G network in virtual area per bandwidth per user.

Reduced dimethyl sulfoxide concentrations successfully cryopreserve human hematopoietic stem cells with multi-lineage long-term engraftment ability in mice

Background aimsThe cryopreservation of hematopoietic stem cells (HSCs) in dimethyl sulfoxide (DMSO) is used widely, but DMSO toxicity in transplant patients and the effects of DMSO on the normal function of cryopreserved cells are concerns. To address these issues, in vitro and clinical studies have explored using reduced concentrations of DMSO for cryopreservation. However, the effect of reducing DMSO concentration on the efficient cryopreservation of HSCs has not been directly measured. MethodsCryopreservation of human bone marrow using 10%, 7.5% and 5% DMSO concentrations was examined. Cell counting, flow cytometry and colony assays were used to analyze different cell populations. The recovery of stem cells was enumerated using extreme limiting dilution analysis of long-term multi-lineage engraftment in immunodeficient mice. Four different methods of analyzing human engraftment were compared to ascertain stem cell engraftment: (i) engraftment of CD33+ myeloid, CD19+ B-lymphoid, CD235a+ erythroid and CD34+ progenitors; (ii) engraftment of the same four populations plus CD41+CD42b+ platelets; (iii) engraftment of CD34++CD133+ cells; and (iv) engraftment of CD34++CD38− cells. ResultsHematopoietic colony-forming, CD34++/+, CD34++CD133+ and CD34++CD38− cells were as well preserved with 5% DMSO as they were with the higher concentrations tested. The estimates of stem cell frequencies made in the xenogeneic transplant model did not show any significant detrimental effect of using lower concentrations of DMSO. Comparison of the different methods of gauging stem cell engraftment in mice led to different estimates of stem cell numbers, but overall, all measures found that reduced concentrations of DMSO supported the cryopreservation of HSCs. ConclusionCryopreservation of HSCs in DMSO concentrations as low as 5% is effective.

Plasticity in the hippocampal formation of shorebirds during the wintering period: Stereological analysis of parvalbumin neurons in Actitis macularius.

The number of parvalbumin neurons can be modified by social, multisensory, and cognitive stimuli in both mammals and birds, but nothing is known about their plasticity in long-distance migratory shorebirds. Here, in the spotted sandpiper (Actitis macularius), we investigated the plasticity of parvalbumin neurons of two brain areas during this species' wintering period at a lower latitude. We compared individuals in a nonmigratory rest period (November-January) and premigration (May-July) period. We used parvalbumin as a marker for counting a subpopulation of inhibitory neurons in the hippocampal formation (HF), with the magnocellular nucleus of the tectal isthmus (IMC) as a control area. Because the HF is involved in learning and memory and social interaction and the IMC is essential for control of head, neck, and eye movements, we hypothesized that parvalbumin neurons would increase in the HF and remain unchanged in the IMC. We used an optical fractionator to estimate cell numbers. Compared with the nonmigratory rest birds, parvalbumin neuron count estimates in the premigration birds increased significantly in the HF but remained unchanged in IMC. We suggest that the greater number of parvalbuminergic neurons in the HF of A. macularius in the premigration period represents adaptive circuitry changes involved in the migration back to reproductive niches in the northern hemisphere.

The Linear Model of Saccharomyces cerevisiae Turbidity in Liquid Media

The study aims to investigate the relationship between the turbidity and density or the total suspended inorganic particles have been obtained many models. The live cells as the homogeneous particle are presumed to cause turbid in liquid media, and that has a linear relationship that can be utilized on the cell counting. The method for the term of clouded liquid form is the measurement based on the reflection and scattered of light, i.e., the turbidimetry. Knowledge attainment of microbial cell counting should be answered how many Nephelometric Turbidity Unit of the one cell. We work to obtain a turbidity model of cells in water-based media for the estimation of cell numbers. This paper aims to construct the computational structure on the turbidity modeling of Saccharomyces cerevisiae in pure water and to test a consistent model in liquid nutrients medium. The modeling was performed in systematized stages of the diagnostic-analysis-test; the regression assurances, the simulation of the lowest error, and the coefficient value itself of turbidity factors. We constructed an optimal analysis and diagnosis to create a computational structure of cell turbidity modeling. The measurement and stopping bivariate elimination of the simulation is a subsystem of the algorithm of obtaining and testing models. The first mathematics model is a standard curve on turbidimetry, and the second, turbidity mathematics model of cell growth in liquid nutrients medium. Both models have an equal coefficient of cell turbidity. The turbidity coefficient of cell growth time interval in the carbonyl diamide - potato dextrose broth is significant.

Real-time monitoring of extracellular ATP in bacterial cultures using thermostable luciferase.

Adenosine triphosphate (ATP) is one of the most important indicators of cell viability. Extracellular ATP (eATP) is commonly detected in cultures of both eukaryotic and prokaryotic cells but is not the focus of current scientific research. Although ATP release has traditionally been considered to mainly occur as a consequence of cell destruction, current evidence indicates that ATP leakage also occurs during the growth phase of diverse bacterial species and may play an important role in bacterial physiology. ATP can be conveniently measured with high sensitivity in luciferase-based bioluminescence assays. However, wild-type luciferases suffer from low stability, which limit their use. Here we demonstrate that an engineered, thermostable luciferase is suitable for real-time monitoring of ATP release by bacteria, both in broth culture and on agar surfaces. Different bacterial species show distinct patterns of eATP accumulation and decline. Real-time monitoring of eATP allows for the estimation of viable cell number by relating luminescence onset time to initial cell concentration. Furthermore, the method is able to rapidly detect the effect of antibiotics on bacterial cultures as Ampicillin sensitive strains challenged with beta lactam antibiotics showed strongly increased accumulation of eATP even in the absence of growth, as determined by optical density. Patterns of eATP determined by real-time luminescence measurement could be used to infer the minimal inhibitory concentration of Ampicillin. Compared to conventional antibiotic susceptibility testing, the method presented here is faster and more sensitive, which is essential for better treatment outcomes and reducing the risk of inducing antibiotic resistance. Real-time eATP bioluminescence assays are suitable for different cell types, either prokaryotic or eukaryotic, thus, permitting their application in diverse fields of research. It can be used for example in the study of the role of eATP in physiology and pathophysiology, for monitoring microbial contamination or for antimicrobial susceptibility testing in clinical diagnostics.

Experimental evidence on the effects of temperature and salinity in morphological traits of the Microcystis aeruginosa complex

Microcystis aeruginosa complex (MAC) encompasses noxious colonial bloom forming cyanobacteria. MAC representatives bloom in eutrophic freshwater and brackish ecosystems with stagnant water, were temperature and salinity are the main variables modulating their distribution, biomass and toxicity. Cell abundance and biovolume of MAC colonies define regulatory standards for public health. These variables depend upon colony size that in turn changes with environmental conditions. Here, we conducted two series of experiments to evaluate the response of MAC colonies morphological traits (length, volume, mucilage and number of cells) to temperature and salinity. In two series of experiments in the laboratory, we exposed natural MAC communities to three different temperatures (10, 21 and 30 °C) and four salinity levels (0, 5, 10 and 25 ppt) typically found in estuaries. We found that average colony length, volume and mucilage thickness did not change with temperature, but the cell-free space inside the colonies was smaller at the highest evaluated temperature (30 °C). Salinity fostered an increase in colony length, volume and mucilage thickness, while cell-free space diminished, resulting in higher cell density. The number of cells per colony was significantly related to colony size (length and volume) and both, temperature and salinity, affected the parameters of the relationships. Based on present results we propose statistical models to predict cell number per colony based on length and volume and accounting for the effect of salinity and temperature on these traits. This is applicable to ecological studies and to the monitoring of estuarine aquatic environments, by means of a fast and more accurate estimation of cell numbers to define MAC toxic populations early warning systems. A protocol is suggested for its application while the analysis of the interaction of temperature and salinity, as well as the variability in natural environments are objectives for future researches.

A Metric Fractionator for Estimation of Total Cell Number in Paraffin-Embedded Flat or Hollow Organs.

The physical fractionator is a convenient and practical solution for estimation of total cell number in a regulatory toxicology setting because it is insensitive to shrinkage allowing for paraffin processing/embedding and does not require measurement of the reference or organ volume. The principle involves sampling a known fraction of an organ in one or more steps and counting the total number of cells present in the final sample, physical disector section pairs. The total cell number in the organ is estimated by multiplying the cell count in the final fraction by the inverse of the sampling fraction(s). The key feature of the design is that tissue shrinkage due to paraffin processing occurs before the organ is uniformly sampled. Another requirement is that thermal expansion or contraction is avoided during the preparation of disector sections from the individual embedded subsamples, which ensures that the disector sections represent a known constant fraction. This vertical physical fractionator with subsampling is a simple and fast estimator to obtain precise and robust estimates of total cell number in large flat or hollow organs that do not prolong routine necropsy procedures. It is compatible with paraffin processing, avoids exhaustive sectioning, and allows for the collection of routine histopathology sections.

Stereological Estimation of Cholinergic Fiber Length in the Nucleus Basalis of Meynert of the Mouse Brain.

The length of cholinergic or other neuronal axons in various brain regions are often correlated with the specific function of the region. Stereology is a useful method to quantify neuronal profiles of various brain structures. Here we provide a software-based stereology protocol to estimate the total length of cholinergic fibers in the nucleus basalis of Meynert (NBM) of the basal forebrain. The method uses a space ball probe for length estimates. The cholinergic fibers are visualized by choline acetyltransferase (ChAT) immunostaining with the horseradish peroxidase-diaminobenzidine (HRP-DAB) detection system. The staining protocol is also valid for fiber and cell number estimation in various brain regions using stereology software. The stereology protocol can be used for estimation of any linear profiles such as cholinoceptive fibers, dopaminergic/catecholaminergic fibers, serotonergic fibers, astrocyte processes, or even vascular profiles.

Turbidity of Saccharomyces cerevisiae: a proposed cell quantification method

The relation between cell and turbidity (cloudiness) in suspended form is studied intensively to facilitate the estimation of the cell number. A standard curve in turbidimetry has utility to compare the turbidity reading value for the determination of Saccharomyces cerevisiae cell number and using a model. A standard curve is a prospect in this issue for the cell that has an inherent inaccuracy in the method as well as spectrophotometry. This research is conducted to obtain a model for the standard curve using analysis and diagnosis of the linear regression which was measured the relative standard error (SE) value of the dependent variable by simulation of turbidity value and verification of the applied model. We obtained the smallest relative SE from one of the five datasets that resulted in less than 10% of the relative SE on the estimated cell number. Plotting all of the bivariate data resulted in a larger error on the scattered light intensity at the higher cell number value, and this characteristic was named heteroscedasticity. Application of turbidity method for cell enumeration per mL in the media was significant equal and lower coefficient variation than the result of verification by a viable method.

Automated superpixels-based identification and mosaicking of cone photoreceptor cells for adaptive optics scanning laser ophthalmoscope

An automated superpixels identification/mosaicking method is presented for the analysis of cone photoreceptor cells with the use of adaptive optics scanning laser ophthalmoscope (AO-SLO) images. This is an image oversegmentation method used for the identification and mosaicking of cone photoreceptor cells in AO-SLO images. It includes image denoising, estimation of the cone photoreceptor cell number, superpixels segmentation, merging of superpixels, and final identification and mosaicking processing steps. The effectiveness of the presented method was confirmed based on its comparison with a manual method in terms of precision, recall, and F1-score of 77.3%, 95.2%, and 85.3%, respectively.