Microscopy Laboratory Research Articles

Morphological Characteristics of Low-asphalt RAP and Influence of its Reaction Product During Combustion Heating on Regenerated Asphalt

To provide reference for regeneration of the finely separated low-asphalt reclaimed asphalt pavement (RAP), the macroscopic and microscopic laboratory testing and quantum chemical simulation were combined. The morphological characteristics of the basalt and limestone low-asphalt RAP were analyzed. The low-asphalt RAP needs to be dried before regeneration, so the reaction mechanism and product of the asphalt film on its surface during drying were clarified. The product influence on the temperature resistance and fatigue performance of the virgin asphalt was studied, so as to consider their mixing during the subsequent regeneration process. Result shows that the gradation of limestone low-asphalt RAP is coarser and particles are slenderer than basalt with the same particle size grade. The 5-10mm low-asphalt RAP particles are slenderer than those in the 10-20mm grade. In the drying cylinder, asphalt basically generate no new functional groups below 200℃, while the asphalt film on the low-asphalt RAP surface undergoes pyrolysis to produce carbon black with the particle size around 100μm in the combustion heating area. The most easily oxidation or pyrolysis site in asphalt molecule is the carbon atom connected to the benzene ring. The pyrolysis of asphalt molecules to generate free radicals in the local oxygen deficient environment is the prerequisite for the carbon black formation, and pyrolysis requires higher temperature than oxidation because of its higher energy barrier. As the carbon black content increases, the penetration and ductility of asphalt decrease, the softening point increases, the high-temperature performance is enhanced, the low-temperature performance is weakened, and the fatigue performance is not significantly affected. As the particle size of carbon black increases, the decrease range in penetration, ductility and low-temperature performance of asphalt declines, the increase or improvement range of softening point and high-temperature performance enhanced, while the fatigue performance still shows no significant and regular change.

Necrosis area, Tumor Size, and Lymph Node of Breast Cancer

Background: Breast cancer is the leading cause of cancer deaths in women worldwide in 2020. The high mortality rate is often influenced by various reasons, one of which is patients's prognosis. One of the prognostic factors is necrosis area, caused by hypoxic tumor tissue that can affect tumor growth and metastasis. Therefore, this study aims to determine the relationship between necrosis area with tumor size and lymph node metastasis in breast cancer patients. Methods: This study was an observational analytic with a cross-sectional approach. This research was conducted at the Anatomical Pathology Laboratory of Dr Soedarso Pontianak General Hospital and the Microscopy Laboratory of the Faculty of Medicine Tanjungpura University from September until November 2022. The study sample was 55 slides of Formalin-Fixed Paraffin-Embedded (FFPE). The study was conducted by observing the preparations under a microscope and then analyzed using ImageJ and SPSS software. The independent variable in this study was the necrosis area in the tumor microenvironment, and the dependent variables were tumor size and lymph node metastasis in breast tissue. Results: All study samples were female patients with invasive ductal carcinoma. Breast cancer tends to occur in patients aged 48-55 years. The highest percentage of necrosis area was 21% and the most diagnosed breast tumor size and lymph node metastasis were stage T4 and stage N2. Spearman's rho correlation test showed that there was no relationship between necrosis area and tumor size (p = 0.122; r = 0.16) and lymph node metastasis (p = 0.637; r = 0.18). Conclusions: There is no correlation between necrosis area with tumor size and lymph node metastasis of breast cancer patients in the Anatomical Pathology Laboratory at Dr Soedarso Pontianak General Hospital.

A Framework For Reflection: Using Play To Understand The Relationship Between Art And Bioimaging

Abstract Bioimaging experiments are carried out in discrete labs, and artists are rarely granted access. They are an underused resource for artists-researchers. In this article I demonstrate how I as an artist-researcher can contribute to Art--Science initiatives in pharmacological research by working in an Advanced Imaging and Microscopy lab and responding innovatively to current circumstances in bioimaging. I do this through thinking about scientific and artistic interdisciplinary practice in a playful way. Informed by established play theories and practices from the literature I reviewed, studied, and then adapted. I conducted research at The School of Life Sciences at the University of Nottingham and evaluated play as an insightful concept to provoke a reaction to scientific methods.

Three-dimensional microscope skill acquisition: A randomised controlled study comparing two-dimensional laboratory microscope training, video gaming and virtual reality gaming

IntroductionFine microsurgical motor skill acquisition can be challenging. With increasing technological innovation the method of microsurgical skills acquisition may change. Studies show that laboratory-based microsurgical training programs on a 2D microscope significantly improves novice’s microsurgical skill acquisition. However, are these skills transferable to a 3D microscope or is gaming agility more important? We present a randomised control trial of three interventions, namely laboratory tabletop microscope training (LM), high-fidelity video gaming (Sony PlayStation 4 console) (VG) and high-fidelity virtual reality gaming (Sony PlayStation VR console) (VR) versus a control group. MethodsForty novice medical students were block randomised to four groups: control (no intervention) n=10, LM n=10, VG n=10 and VR n=10. Participants performed chicken femoral artery anastomosis using the Aesculap Aeos® 3D microscope platform at baseline and again after intervention. Performance was evaluated using a modified structured assessment of microsurgery skills (mSAMS) score, time taken to complete anastomosis and time for suture placement by two blinded independent assessors. ResultNo statistically significant difference was noted between the groups at baseline. There was a statistically significant improvement in the LM arm between baseline and post-training for mSAMS score and time for suture placement. In the VG, VR and control groups no statistically significant difference was observed. ConclusionOur study demonstrates that during early microsurgical training, an intense laboratory-based microsurgical training program significantly improves novice’s anastomotic performance on a 2D microscope, and these skills are transferable when a 3D anastomosis is carried out. However, focussed gaming had no significant effect, and the results were akin to the non-intervention group.

PyFMLab: Open-source software for atomic force microscopy microrheology data analysis

Background Atomic force microscopy (AFM) is one of the main techniques used to characterize the mechanical properties of soft biological samples and biomaterials at the nanoscale. Despite efforts made by the AFM community to promote open-source data analysis tools, standardization continues to be a significant concern in a field that requires common analysis procedures. AFM-based mechanical measurements involve applying a controlled force to the sample and measure the resulting deformation in the so-called force-distance curves. These may include simple approach and retract or oscillatory cycles at various frequencies (microrheology). To extract quantitative parameters, such as the elastic modulus, from these measurements, AFM measurements are processed using data analysis software. Although open tools exist and allow obtaining the mechanical properties of the sample, most of them only include standard elastic models and do not allow the processing of microrheology data. In this work, we have developed an open-source software package (called PyFMLab, as of python force microscopy laboratory) capable of determining the viscoelastic properties of samples from both conventional force-distance curves and microrheology measurements. Methods PyFMLab has been written in Python, which provides an accessible syntax and sufficient computational efficiency. The software features were divided into separate, self-contained libraries to enhance code organization and modularity and to improve readability, maintainability, testability, and reusability. To validate PyFMLab, two AFM datasets, one composed of simple force curves and another including oscillatory measurements, were collected on HeLa cells. Results The viscoelastic parameters obtained on the two datasets analysed using PyFMLab were validated against data processing proprietary software and against validated MATLAB routines developed before obtaining equivalent results. Conclusions Its open-source nature and versatility makes PyFMLab an open-source solution that paves the way for standardized viscoelastic characterization of biological samples from both force-distance curves and microrheology measurements.

Enhancing Efficiency and Collaboration in Research-Driven Microscopy Laboratory

Enhancing Efficiency and Collaboration in Research-Driven Microscopy Laboratory

Microscopic and statistical evaluation of the marginal defects of composite restorations: in vitro studies.

This study aimed to evaluate the quality of dental restorations using laboratory microscopic techniques, focusing on the positioning of restorations relative to cavity edges and identifying common errors, such as incomplete or excessive coronal restorations. A total of 41 extracted lateral teeth, previously treated in vivo with Class I or II composites, were analyzed. Thirty-three of these teeth were sectioned medio-distally or buccal-orally and examined under a research laboratory microscope. Marginal areas were measured using eyepieces with a graduated scale and calibration slide, and a frequency statistical analysis was conducted. The analysis revealed that the teeth had fractious edges, deficient marginal closure, excess composite, incorrectly restored occlusal cusps, and marginal adaptation errors, with approximately half of these errors involving excess material. It was observed that restoration procedures in distal areas are particularly challenging, and clinical errors with excess material occur more frequently than those with a deficit. These findings underscore the need for improved techniques and precision in dental restorations to minimize such errors.

Synergistic Fermentation of Pichia kluyveri and Saccharomyces cerevisiae Integrated with Two-Step Sugar-Supplement for Preparing High-Alcohol Kiwifruit Wine.

Wild yeast suitable for kiwifruit wine fermentation was isolated and purified, and the fermentation process was optimized to increase the alcohol content of the kiwifruit wine. Pichia kluyveri was isolated from kiwifruit pulp by lineation separating, screened by morphological characteristics in Wallerstein Laboratory Nutrient Agar (WL) medium and microscope observation, and further identified by 26S rDNA D1/D2 domain sequence analysis. Taking alcohol content and sensory evaluation as two indexes, the fermentation condition for kiwifruit wine was optimized by single factor and response surface experiment. The optimal fermentation conditions were optimized as follows: the fermentation temperature was at 24 °C, the initial pH was 3.8, the sugar dosage in second step was 8% (w/w), and the inoculating quantity of Pichia kluyveri and Saccharomyces cerevisiae was 0.15 g/L at equal proportion. Under these optimal conditions, the maximum estimated alcohol content was 15.6 vol%, and the kiwifruit wine was light green in color with strong kiwifruit aroma and mellow taste.

Reducing costs of Giardia spp. cyst enumeration using machine learning-based systems

Abstract Giardia spp. cyst enumeration is a laboratory procedure that is frequently required in water treatment-related research. Currently, detection conducted by experts using fluorescence microscopy on samples stained with specific markers for Giardia spp. cysts is still the standard method, despite its high costs limiting its usage worldwide and, ultimately, hindering waterborne analyses in low-income countries. We present an approach based on darkfield imaging and machine learning to reduce costs associated with Giardia spp. cyst enumeration and the lack of experts. Automated counts were compared to manual counts, achieving an average sensitivity (SE) rate of 88%, specificity (SP) of 100% and accuracy of 88% across a wide range of cyst concentrations. By using machine learning in conjunction with darkfield microscopy, a low-cost illumination technique that can be easily integrated into standard laboratory microscopes, we have significantly reduced the costs associated with Giardia spp. cyst detection, all while still maintaining the SE and SP of fluorescence microscopy. Based on the findings, the proposed system has the potential to be a useful tool to enumerate Giardia spp. cyst suspensions. It can be accessed by virtually any microbiology laboratory as it is consumable-free and expert-independent.

Evaluation of Local Common Bean (Phaseolus vulgaris L.) Genotypes for Resistance to Fungal Diseases in the Bimodal-rainfall Forest Zone of Cameroon

The general objective of the present study was to evaluate the susceptibility to fungal diseases of some bean collections in the bimodal rainfall forest zone of Cameroon. To this end, 12 bean collections from Foumbot were grown at the University of Yaoundé I in a completely randomized block design without phytosanitary treatments. Symptoms identified on the leaves of diseased plants were used to characterize the fungal agents in the laboratory using an identification key. Incidence and severity were assessed, as well as the number of pods and seeds in plants under fungal pressure. Following the evolution of symptoms, all collections showed fungal disease attacks at different rates. Macroscopic and microscopic laboratory observations identified Sclerotinia sclerotiorum, Uromices appendiculatus and Fusarium solani as the fungal agents responsible for these symptoms. The incidence of disease caused by F. solani was lower in the NJBPRNV (16.67%) and NJBV (22.22%) collections, as was severity (3.33% and 4.44%). On the other hand, the LGRTE (47.06%) and GGRBE (44.44%) LGRV (43.75%) collections showed high incidences of disease caused by S. sclerotiorum, U. appendiculatus and F. solani respectively. Considering the number of seeds, the KPGPV (403.2 seeds) and KPGPTV (350.4 seeds) collections recorded the highest number of seeds, compared with the GGRBE (15.93 seeds); LGRTE (23.87 seeds); GGRBTE (25.53 seeds); LGRV (42.4 seeds) and GGRTE (52.2 seeds) collections. However, the NJBPRNV collection requires special attention in varietal creation, given its greater resistance to the fungal diseases identified.

Correlation of light and electron microscopic morphometric parameters of glomerular capillaries with serum creatinine and proteinuria

ABSTRACT Waste products in the bloodstream are filtered by the glomerular capillaries in the kidneys and excreted into the urine. When making a differential diagnosis of kidney diseases, structural assessment of glomeruli using histological, ultrastructural, and immunological studies is crucial. This study assessed the microscopic and ultrastructural morphometric parameters of glomerular capillaries and examined their correlation with serum creatinine and proteinuria. A total of 60 kidney biopsy cases received by the transmission electron microscope (TEM) laboratory for diagnosis were included in the study. Toluidine blue stained 300 nm thick sections of TEM tissue blocks were scanned for glomerular morphometry by a whole slide imaging system, and the estimation of Bowman’s capsule (BC) area, glomerular capillary lumen diameter (GCLD), glomerular capillary density (GCD), glomerular capillary surface area density (GCSA), and percentage of glomerular capillary lumen space (%GCLS) was performed with QuPath software. TEM images of 70 nm thick sections were used for the evaluation of endothelial fenestration diameter (EFD), glomerular basement membrane (GBM) thickness, and podocyte foot process (PFP) effacement. Proteinuria and serum creatinine showed positive correlations with GBM thickness and PFP effacement. Negative correlations of serum creatinine were observed with EFD, %GCLS, and GCSA. Hence, glomerular filtration is greatly affected by the total area of the glomerular capillary surface and structural changes of GBM. Reduction of glomerulus filtration due to foot process effacement and thickening of GBM results in damage to the filtration barrier leading to the leakage of plasma protein into urine.

Creating Efficient Workflows for Electron Microscopy Laboratories with Automated Specimen Preparation

Abstract Electron microscopy (EM) is essential to the biological and biomedical sciences and clinical diagnostic pathology. Electron microscopy specimen preparation is laborious and time-consuming, with transmission EM (TEM) chemical preparation typically requiring 1–3 days, while volume electron microscopy (vEM) takes 3–5 days of tedious manual reagent exchanges every few minutes or hours. This places a considerable burden on laboratory scientists with the ongoing demand for TEM, and the rapidly growing demand for vEM due to its potential to revolutionize structural biology, connectomics, and related fields. This burden is exacerbated by a shortage of trained electron microscopy scientists as current staff retire, and few enter the workforce. This report provides four case studies to illustrate how automated and faster specimen preparation workflows using mPrep™ Automated Specimen Processors (ASP-1000™ and ASP-2000™, Microscopy Innovations, LLC) free electron microscopy staff in academic and pre-clinical research labs and a clinical pathology laboratory.

Особенности выявления костных включений в мясных изделиях микроструктурными методами.

Mechanically separated poultry meat (MSPM), being a valuable food raw material, is widely used in the meat industry. Therefore, its detection in finished products is an urgent task in controlling their quality and composition. The article presents the results of the application of histochemical staining methods to identify bone fragments as the main indicator of the presence of MSPM in meat products. To ensure accuracy and first of all, comparability of results of histological investigations, it is important that all laboratories adhere to the principles of the microscopic laboratory practice and use the uniform methodology for preparation of histological specimens and interpretation of obtained results.

Analysis of Barbie medical and science career dolls: descriptive quantitative study.

To identify Barbie brand dolls that had medicine and science themed professions in comparison with other career dolls and to determine their accuracy in meeting clinical and laboratory safety standards. Descriptive quantitative study. Visual and data analysis of web searches. To identify the kinds of medical and scientific subspecialties that the Barbie dolls (and a comparison doll group) worked in; and to determine whether these medical professional and scientist dolls met laboratory and clinical safety standards. Additional data about doll demographics (ie, age, ethnic group, and sex) were also collected. 92 Barbie brand dolls were analyzed: doctor (n=53), scientist (n=10), science educator (n=2), nurse (n=15), dentist (n=11), and paramedic (n=1). 65 non-Barbie brand dolls were also analyzed for comparison purposes: doctor (n=26), scientist (n=27), nurse (n=7), dentist (n=2), engineer (n=2), and magnetic resonance imaging (MRI) technician (n=1) dolls. Barbie brand medical professional dolls (n=80) largely treated children (66%, n=53/80), with only three (4%) medical professional dolls being directly depicted working with adults. Of the 12 scientist Barbie brand dolls, none met all proper personal protective equipment requirements related to hair and clothing. Barbie brand dolls often came with items, such as laboratory coats, microscopes, stethoscopes, and glasses, that children stereotypically associate with doctors and scientists. While comparison dolls offered a wider range of age and ethnic groups than the Barbie doll group did, the dolls similarly struggled to portray a wide range of medical and scientific subfields and most comparison dolls did not wear proper personal protective equipment. Medicine and science themed dolls help to inspire tomorrow's medical professionals and scientists. All toy companies should ensure that future medical professional and scientist dolls meet clinical and laboratory safety standards and diversify the types of medical and scientific professions represented (especially among male dominated fields). For young girls' sakes as much as her own, Barbie must keep shattering glass ceilings.

PyFMLab: Open-source software for atomic force microscopy microrheology data analysis.

Atomic force microscopy (AFM) is one of the main techniques used to characterize the mechanical properties of soft biological samples and biomaterials at the nanoscale. Despite efforts made by the AFM community to promote open-source data analysis tools, standardization continues to be a significant concern in a field that requires common analysis procedures. AFM-based mechanical measurements involve applying a controlled force to the sample and measure the resulting deformation in the so-called force-distance curves. These may include simple approach and retract or oscillatory cycles at various frequencies (microrheology). To extract quantitative parameters, such as the elastic modulus, from these measurements, AFM measurements are processed using data analysis software. Although open tools exist and allow obtaining the mechanical properties of the sample, most of them only include standard elastic models and do not allow the processing of microrheology data. In this work, we have developed an open-source software package (called PyFMLab, as of python force microscopy laboratory) capable of determining the viscoelastic properties of samples from both conventional force-distance curves and microrheology measurements. PyFMLab has been written in Python, which provides an accessible syntax and sufficient computational efficiency. The software features were divided into separate, self-contained libraries to enhance code organization and modularity and to improve readability, maintainability, testability, and reusability. To validate PyFMLab, two AFM datasets, one composed of simple force curves and another including oscillatory measurements, were collected on HeLa cells. The viscoelastic parameters obtained on the two datasets analysed using PyFMLab were validated against data processing proprietary software and against validated MATLAB routines developed before obtaining equivalent results. Its open-source nature and versatility makes PyFMLab an open-source solution that paves the way for standardized viscoelastic characterization of biological samples from both force-distance curves and microrheology measurements.

Nitrofurantoin Susceptibility Profile on Bacterial Isolates from Urinary Tract Infection in Patients with Diabetes Mellitus

Background: Patients with diabetes mellitus have a higher risk of urinary tract infection (UTI) incidence than those without. One of the first-line therapy for UTI is nitrofurantoin. The emergence of drug-resistant UTIs is increasing both in community and healthcare setups. Purposes: Determine the nitrofurantoin susceptibility profile to bacteria causing UTI in diabetic patients. Method: A cross-sectional study was conducted at the Microscopic Laboratory, Faculty of Medicine, Universitas Tanjungpura, Pontianak, from February to November 2019. The antibiotic susceptibility testing was performed by the Kirby-Bauer disk diffusion method on 22 bacterial isolates. The samples were collected on a sterile urine pot, cultured on agar, and identified by biochemistry test, and bacterial isolates were stored at 2-8oC. The bacterial isolates were sub-cultured 24h before the susceptibility test. The susceptibility testing used a 300 µg nitrofurantoin disk. The diameter of zone inhibition was measured and classified based on Clinical and Laboratory Standard Institute guidelines. Result: The results showed that Escherichia coli was 41.67% susceptible (5/12), Shigella sp. was 100% intermediate (1/1), and Enterobacter aerogenes (3/3), Pseudomonas aeruginosa (1/1), Klebsiella sp (1/1) were 100% resistant. Conclusion: The majority of the causative bacteria for UTI in diabetic patients are resistant to nitrofurantoin, suggesting the use of nitrofurantoin should be reconsidered as an empirical antibiotic in Pontianak. Further study using a larger population should be conducted to describe a more extensive antibiotic susceptibility profile of diabetic patients with UTI in Pontianak.

Indigenous artefacts and oral stories

This article presents the results of studies conducted by Canadian academics in collaboration with Sturgeon Lake and Pelican Narrows First Nations communities (Saskatchewan, Canada). The objectives of the project were: (1) developing a research ethics protocol for collecting, studying, and preserving indigenous artefacts; (2) measurements of chemical compositions of artefacts; (3) collecting oral stories of Elders. Within the project, two workshops were organized in Pelican Narrows and Sturgeon Lake. Post-secondary students were trained to work on the project. The laboratory measurements of chemical compositions of artefacts were conducted at the Scanning Electron Microscope Laboratory (University of Alberta), and the Saskatchewan Isotope Laboratory (University of Saskatchewan). The carbon-dating measurements were carried out at André E. Lalonde Accelerator Mass Spectrometry Laboratory (University of Ottawa). The statistical analysis of chemical compositions was conducted in order to test the provenance similarities of artefacts. The project was supported by the Department of Canadian Heritage.

METHOD OF RESTORATION AND REINFORCEMENT OF TEETH WITH SIGNIFICANT DESTRUCTION OF THE CROWN OF THE TOOTH

Background: One of the ways to improve the quality of restoration of the crown part of the tooth is to add a reinforcing component that increases the strength and durability of the restoration.
 Aim:Propose a new method for restoring extensive defects in hard tissues of teeth with additional reinforcement
 Materials and methods: In the course of the work, 100 chewing anatomical and functional teeth removed according to various indications were used. Carious cavities of all teeth after removal were prepared for class 1 G.V. Blаck cavities. The wall thickness was measured using a dental micrometer, the wall thickness of half of the teeth (40) was more than 3 mm, half (60) - less than 3 mm, on the basis of which it was calculated how the load would be distributed in the tooth, based on this, the optimal installation method was determined threads with composite in the tooth cavity. We used: dental adhesive Single Bond Universal in separate packages, dental composite material 3M ESPE Filtek Ultimate Universal Restorative, thread with composite material (light-curing dental fiber formed CATERPILLAR), also during the work we used a dental micrometer, laboratory electrical equipment: testing machine, hardness tester, Shimadzu trademark, LEDEX dental light curing lamp, model WL-070, Carl Zeiss Primo Star laboratory microscope, emission scanning electron microscope Merlin (Zeiss).
 Conclusion:The "Method of restoration of molars in case of significant destruction of the crown part of the tooth" has been developed (RF Patent No. 2783060). It is proposed as a method of restoration of extensive defects of the crown part of the tooth using a composite tape as a reinforcing component. When analyzing the results of the study on the Shimadzu bursting machine, it was revealed that restorations carried out using a composite thread are more durable and, accordingly, durable than similar therapeutic restorations of the shape and function of the tooth carried out without using a composite thread.
 Results: In a laboratory test for maximum compression and stress, teeth restored using a combination of floss with composite and composite material have shown significantly greater resistance than teeth restored with composite material alone.

Occurrence Characteristics of Microplastics in Baiyangdian Lake Water and Sediments

In order to explore the occurrence characteristics of microplastics in the freshwater environment of Baiyangdian Lake in China, ten overlying water samples and ten sediment samples were collected in Baiyangdian Lake of Hebei Province in October 2021, and the abundance distribution, shape, particle size, and polymer type of microplastics in the samples were identified using laboratory pretreatment, microscope observation, and laser infrared spectroscopy. The sedimentation law of microplastics at the overlying water-sediment interface was studied using the Stokes sedimentation formula, and their pollution characteristics and potential sources were analyzed. The abundances of microplastics in the overlying water and sediments in Baiyangdian Lake ranged from 474-19382 n·m-3 and 95.3-29542.5 n·kg-1, respectively, with an average value of 6255.4 n·m-3 and 11088 n·kg-1. The main polymer of the microplastics in the overlying water was polyethylene terephthalate[PET, (17.20±0.26)%], and the microplastics in the sediments were mainly chlorinated polyethylene[CPE, (46.11±1.30)%]. The sedimentation velocities of microplastics in the sedimentation zone ranged from 0.0793-111.7547 mm·s-1. The particles with larger particle size had higher sedimentation velocity and easily settled and remained in the sediments. The main sources of microplastic pollution in the study area were the discharge of textile fibers from washing wastewater and the wear and tear of ship paint, ship rubber, and building materials.

Comparative study of infection control practices in DOTS/sputum microscopy centre in two different districts of state of Uttarakhand and Uttar Pradesh of India

Background & ObjectiveTuberculosis (TB) is endemic in India. TB is transmitted through droplet infection and the transmission occurs when a person inhales droplet nuclei containing of Mycobacterium tuberculosis. Infection control practices play a major role in controlling the TB infection in healthcare settings and further prevents TB infection in the HCWs. The aim of the study is to conduct the cross sectional study for infection control practices in DOTS cum Sputum Microscopy Centre's under NTEP in Ghaziabad (Uttar Pradesh) & Dehradun (Uttarakhand) districts with the objective to assess the compliance of infection control measures by HCWs in DOTS cum Sputum Microscopy Centre's and to suggest the suitable measures and/or model to reduce the transmission of infection to the HCWs and to the community at large. Materials & methodsThe cross sectional study is conducted for two years in two districts of different state having high burden of TB disease in UP and low burden of disease in UK state. All DOTS cum Sputum Microscopy centres of both selected districts i.e. 100% sample size are covered in the study. ResultsHand washing is the most efficient and cost-effective practice for prevention and control of infection. In Dehradun district 66.66% (12) centers and in Ghaziabad district 57.14% (16) centers have adequate hand washing facility available at DOTS and sputum microscopy lab. Unavailability of adequate PPE will lead to the infection. In Dehradun district, 55.56% (10) centers have adequate PPE available whereas in Ghaziabad District 21.43% (6) centers have adequate PPE available. Training on infection prevention and control for HCWs are provided in 27.78% (5) DOTS/sputum microscopy center in Dehradun whereas none of the DOTS/sputum microscopy center in Ghaziabad district are given training on infection prevention & control for HCWs in last one year. Adequate ventilation plays an important role in transmission of TB/MDR TB or any respiratory infection. HCWs working in DOTS/Sputum microscopy center are at risk to contact the TB/MDR TB infection if there is no proper ventilation in their working places. In 33.33% (6) DOTS/sputum microscopy center in Dehradun & 28.57% (8) in Ghaziabad district have adequate ventilation. Layout of DOTS room and for sputum microscopy center are suggested to reduce the risk of transmission of TB/MDR-TB and other respiratory pathogens amongst HCWs who are working in DOTS cum sputum microscopy center. ConclusionDOTS cum Sputum Microscopy Centers of both districts in different states are having deficient infection control practices. Staff is not adequately trained in infection prevention and control practices.