Staining Properties Research Articles

MO264COMPLEMENT PATHWAY MIGHT HAVE A ROLE IN FOCAL SEGMENTAL GLOMERULOSCLEROSIS: A RETROSPECTIVE SINGLE CENTER STUDY OF 15 YEARS

Abstract Background and Aims The role of complement in focal segmental glomerulosclerosis (FSGS) is an area of interest and C4d staining could indicate complement related renal damage. We investigated detailed C4d staining properties in native kidney biopsies and possible relation to clinical features. Method We retrospectively evaluated the renal biopsies of 114 patients diagnosed with FSGS within last 15 years. C4d expressions examined in glomeruli (mesangial and/or capillary wall, vascular pole, sclerotic areas), tubular region (basement membrane, adsorbtion droplet) and vascular areas (arteriols and arteries) via immunohistochemistry. A novel glomerular C4d score (G-C4d-S) was achieved on the basis of the localization, pattern, extent and intensity of the C4d expression (min-max, 0-13). Results Of the patients (56 females, mean age 43±14 years) with a median follow-up time of 35±3 months, mean proteinuria, eGFR, and albumin level were 4984 mg/day, 72.2 ml/min/1.73m2, 3.56 g/dL, respectively. Median G-C4d-S was 6 (IQR, 4-7) and it was negatively correlated with serum creatinine at diagnosis (r=-0.21, p=0.02). Glomerular staining (both focal and diffuse, higher than moderate in intensity) was positive in 78 (68.4%) of the patients. C4d on glomerular sclerotic area was positive in 43 (37%) patients and it was associated with lower eGFR at diagnosis. Forty five patients achieved remission during the follow-up. Among the pathological features only glomerular C4d staining was associated to remission (p=0.02). There were 20 (18.7%) patients who need renal replacement theraphy (RRT) and 7 deaths (6.1%) at the end of the cohort. Lower rate of C4d staining on tubular adsorption droplets and arteriols/arteries were found to be associated need for RRT (p=0.013, p=0.012, respectively). There were no significant relationship between mortality and C4d staining features. Conclusion In conclusion, we noted that significant number of patients had positivie C4d on glomeruli, arteiroles and tubular area. We indicated that C4d staining at diagnosis could help to distinguish active glomerulonephritis. Additionally, it seems to be essential to examine non-glomerular area of native kidney biopsies, as well.

An improved method for processing chondroprogenitor pellets following chondrogenic differentiation for histology and immunohistochemical staining using agarose

PurposeIn-vitro models of cartilage regeneration based on pellet cultures have been widely used to evaluate chondrogenic potential of the cell of interest and predict probable in-vivo behavior. However, pellet processing is a major challenge during handling (due to small size and possible damage to structural contour following sectioning and staining). The present study aimed to utilize human articular cartilage derived chondroprogenitors to assess if agarose-encapsulation of pellets prior to paraffin processing enable easier handling without affecting tissue morphology, glycosaminoglycan staining and immunohistochemical analysis of Collagen type II protein. MethodsPassage 3 chondroprogenitors (n = 3) were evaluated for MSC markers using flow cytometry and subjected to chondrogenic differentiation as pellets cultures. Post-differentiation, the pellets were subjected to either: a) paraffin embedding, b) agarose encapsulation followed by paraffin embedding or c) agarose encapsulation followed by cryosectioning. All sections were subjected to histological staining for glycosaminoglycan uptake: Alcian blue, Safranin O (Bern score) and Toluidine blue with immunohistochemical processing for collagen type II protein deposition. ResultsWith respect to staining and structural integrity, comparable uptake was seen in both paraffin sections and agarose embedded sections while the latter exhibited notably uniform pellets with distinct marginal demarcation. Although plain paraffin and agarose encapsulated sections demonstrated equivalent staining as represented by comparable Bern scores, glycosaminoglycan uptake, and Collagen type II deposition, cryosections exhibited significantly poor staining properties. ConclusionAgarose encapsulation of differentiated pellets prior to routine paraffin embedding, eases handling difficulties whilst maintaining structural integrity with optimal staining outcomes.

Bacterial Flora Isolated From Dried Fishes Sold at Retail Markets Within Dhaka City Corporation of Bangladesh

Along with other contaminants, bacterial contamination in the dried fishes is a common issue which severely affects the quality of cured fishes. Hence, the present study was aimed to isolate and identify different bacterial flora contaminating different dried fishes, sold at different retail markets within Dhaka Metropolitan city, Dhaka, Bangladesh. A total of 25 different dried fishes were collected from Dhaka North city corporation area (Town Hall kacha bazar, Krishi market, and Mohammadpur kacha bazar) and Dhaka South city corporation area (New market kacha bazar, and Jatrabari chowrasta bazar). Each sample was divided into three regions (head, body, and tail) and bacterial isolation and identification was done by studying their cultural, staining and biochemical properties. Of the 25 dried fishes, Escherichia coli, Bacillus spp., Staphylococcus spp., Salmonella spp. and unidentified bacteria isolation rates were 44%, 56%, 80%, 48%, and 24%, respectively. The presence of E. coli and Salmonella spp. in various dried fish samples could be considered as indication of lack of hygienic condition during dried fish processing. Confirmation through molecular detection methods, pathogenicity, and antibiogram of the isolated bacteria could be included for future study.
 Res. Agric., Livest. Fish.8(1): 109-115, April 2021

Case Report: Polyvinylpyrrolidone deposition disease from repeated injection of opioid substitution drugs: report of a case with a fatal outcome

Background: Intravenous injection of oral opioid substitution drugs (OSD) is widespread among injecting drug users. Several OSDs contain the polymer polyvinylpyrrolidone (PVP) as an excipient. Parenterally administered PVP of high molecular weight may accumulate in tissues and organs. This phenomenon was first described in the 1950s, when PVP was utilised in medication for parenteral use. We report a case of an opioid-addicted patient with extensive PVP–deposition caused by repeated injections of OSDs. Case presentation: A 30-year-old male drug addicted patient in opioid substitution therapy (OST) was repeatedly referred to his local hospital in a poor general condition. Work-up revealed severe normocytic anaemia, renal insufficiency, pancreas insufficiency and pathological fractures. Biopsies from fractured bones, bone marrow and gastric mucosa showed extensive infiltrates of histiocytes with intracytoplasmic vacuoles. Vacuole content stained slightly bluish in hematoxylin and eosin stain, red in Congo red stain and black in periodic acid methenamine silver stain. The morphological appearance and staining properties were in accordance with the diagnosis of PVP deposition. The patient had been injecting both buprenorphine tablets and a specific methadone syrup for several years. The methadone syrup contained large amounts of high molecular weight PVP, making it the most likely cause of the deposition. His health quickly deteriorated and he died, impaired by multi-organ failure and cachexia, five years after the first diagnosis of PVP-deposition. The autopsy revealed extensive PVP-deposition in all sampled organs and tissues. Conclusions: Histological investigation and the correct identification of PVP in the biopsies led to the discovery of a severe adverse effect from long-standing misuse of a drug. The disseminated PVP deposition likely contributed to multi-organ dysfunction and cachexia with a fatal outcome. The deposited PVP likely originated from repeated injections of a certain methadone syrup.

Morphology and histology of the olfactory organ of two African lungfishes, Protopterus amphibius and P. dolloi (Lepidosirenidae, Dipnoi)

The olfactory organs of two African lungfishes, Protopterus amphibius and P. dolloi, were investigated using a stereo microscope and a compound light microscope and were described anatomically, histologically, and histochemically. Like other lungfishes, these species present the following general features: i) elongated olfactory chamber (OC), ii) anterior nostril at the ventral tip of the upper lip, iii) posterior nostril on the palate of the oral cavity, iv) lamellae with multiple cell types such as olfactory receptor neurons, supporting cells, basal cells, lymphatic cells, and mucous cells (MC), and vi) vomero-like epithelial crypt (VEC) made of glandular epithelium (GE) and crypt sensory epithelium. Some of these features exhibit differences between species: MCs are abundant in both the lamellar and inner walls of the OC in P. amphibius but occur only in lamellae in P. dolloi. On the other hand, some between feature differences are consistent across species: the GE of both P. amphibius and P. dolloi is strongly positive for Alcian blue (pH 2.5)-periodic acid Schiff (deep violet coloration), and positive with hematoxylin and eosin and with Masson’s trichrome (reddish-brown staining), unlike the MCs of the two species which stain dark red with both Alcian blue (pH 2.5)-periodic acid Schiff and Masson’s trichrome but respond faintly to hematoxylin and eosin. The differing abundance of MCs in the two lungfishes might reflect different degrees in aerial exposure of the olfactory organ, while the neutral and acid mucopolysaccharide-containing VEC, as indicated by staining properties of the MCs, is evolutionary evidence that P. amphibius and P. dolloi are the closest living relatives to tetrapods, at least in the order Dipnoi.

Surprise underneath a ptosis – Conjunctival and tarsal plate Amyloidosis presenting as ptosis

Amyloidosis is a spectrum of disorders characterized by the deposition of amyloid , an extracellular protein in an abnormal fibrillar form, with highly characteristic histopathologic staining properties. Eyelid involvement can be associated with both immunologic (primary) and reactive (secondary) amyloidosis.We present a case of unilateral localized conjunctival and tarsal plate amyloidosis, presenting as ptosis. Keywords: Amyloid, Conjunctiva, Tarsus, Ptosis.

Synthesis and Application of Oxidized Trehalose as a Hydrophilic Anti-crease Finishing Reagent for Cotton Fabric

In order to improve the anti-crease property of cotton fabrics, it is necessary to chemically cross-link functional groups in the fabric via anti-crease finishing. However, the large amount of hydroxyl groups in cotton cellulose is consumed during the cross-linking finishing process, which greatly reduces the hydrophilicity of the fabric, and thus, staining of cotton fabrics is greatly reduced. Consequently, the fabric becomes non-conducive to subsequent processing and is not well suited for typical applications of cotton fabric. In this paper, a new type of anti-crease finishing reagent for cotton fabric was prepared by using sodium periodate to oxidize Trehalose. This new finishing improved the anti-crease property, and the hydrophilicity and staining of the fabric. The optimum preparation process of oxidized trehalose (OTr) was determined by analyzing the influence of initial pH, molar ratio of trehalose to sodium periodate, and the reaction time on the amount of aldehyde groups formed. The structure of OTr was characterized by FTIR, 13C NMR, MALDI-TOF mass, and the reaction process of OTr was analyzed. The anti-crease property, stiffness, whiteness, tensile strength, hydrophilicity and staining property of fabrics treated with OTr was compared to conventional anti-crease finishing reagents. Results show that the anti-crease property and stiffness of the OTr-treated fabric were significantly improved. The crease recovery angle reached 254°. The whiteness, tensile strength, hydrophilicity and staining of the fabric remained closer to that of the non-treated fabric, as compared to other anti-crease treatments.

H3S28PAntibody Stainingof Okinawan Oikopleura dioicaSuggests the Presence of Three Chromosomes.

Oikopleura dioica is a ubiquitous marine zooplankton of biological interest owing to features that include dioecious reproduction, a short life cycle, conserved chordate body plan, and a compact genome. It is an important tunicate model for evolutionary and developmental research, as well as investigations into marine ecosystems. The genome of north Atlantic O. dioica comprises three chromosomes. However, comparisons with the genomes of O. dioica sampled from mainland and southern Japan revealed extensive sequence differences. Moreover, historical studies have reported widely varying chromosome counts. We recently initiated a project to study the genomes of O. dioicaindividuals collected from the coastline of the Ryukyu (Okinawa) Islands in southern Japan. Given the potentially large extent of genomic diversity, we employed karyological techniques to count individual animals' chromosomes in situ using centromere-specific antibodies directed against H3S28P, a prophase-metaphase cell cycle-specific marker of histone H3. Epifluorescence and confocal images were obtained of embryos and oocytes stained with two commercial anti-H3S28P antibodies (Abcam ab10543 and Thermo Fisher 07-145). The data lead us to conclude that diploid cells from Okinawan O. dioicacontain three pairs of chromosomes, in line with the north Atlantic populations. The finding facilitates the telomere-to-telomere assembly of Okinawan O. dioica genome sequences and gives insight into the genomic diversity of O. dioica from different geographical locations. The data deposited in the EBI BioImage Archive provide representative images of the antibodies' staining properties for use in epifluorescent and confocal based fluorescent microscopy.

Evaluation of Roselle (Hibiscus sabdariffa L.) Calyx Extract Stability for Intestinal Nematode Egg Staining

Roselle (Hibiscus sabdariffa L.) calyx extract is rich in anthocyanin and it has been widely used as a natural colouring agent, for example as an alternative dye for eosin 2% in intestinal nematode egg staining. However, as a natural colour, the temperature and storage time are often altered anthocyanin activity. Therefore, the objective of this study was to investigate stability of Roselle calyx extract using various time and temperature storage and its staining properties on the intestinal nematode eggs. The Roselle calyx was extracted using maceration technique for ± 3 days and then made 80% concentration of extract. Extracted is stored within 1, 7, 14, and 21 evaluation days in room temperature (25°C) and cold temperature (4°C) for extract characterization, absorption of anthocyanin, pH, and staining observation. The result showed that natural extract from Roselle calyx stained intestinal nematode eggs and stable within 21 days storage in cold temperature (4°C).

Comparison between Homemade Stain Remover and Commercial Stain Remover for Textiles

Stain remover is used to remove or masks stain from textiles. Two types of textile stain removers were compared in this study; store bought and home prepared types. Due to the environmental and health issues associated with commercial household cleaners, as well as costly, there have been attempts by consumer especially housewives to prepare cleaning products by using materials which can be found in the kitchens. Hence, the main objective of this study is to compare the effectiveness of home prepared textile stain remover with commercial stain remover by assessing the stain properties on cotton and polyester fabrics. Two different brands of stain remover and easily found materials were applied on these two fabrics. The stains on the fabrics were then assessed according to AATCC Test Method 130 and by using chromameter for the intensity of the stain after cleaning. The results showed that, for the cotton fabric, the most effective stain remover is Commercial Brand 1. Commercial Brand 1 and vinegar with baking soda demonstrated an encouraging effect on polyester fabric. The commercial textile stain remover shows great cleaning effect on both cotton and polyester, while home prepared stain remover has limited ability to clean the stains on cotton. However, its cleaning effect on polyester is equivalent to commercial product.

Brief descriptions of the principles of prominent methods used to study the penetration of materials into human hair and a review of examples of their use.

Consumers are attracted to the latest fashion trends and different looks. This drives the search for novel hair treatments. Some chemicals present in hair treatment products can penetrate the hair shaft. These materials can either nourish or injure the hair cortex. Different techniques have been used to investigate the mechanism of molecule penetration and the conditions under which penetration occurs. This article reviews the techniques applied for this purpose. Various microscopy techniques are used to capture clear and colourful images to determine the diffusion pathways and the exact location of the molecules under study. However, the laborious sample preparation often leads to sample destruction since cross-sectioning is often required. While various other techniques have been successfully used for investigating the penetration methods, most of these require different amounts of work to be put in for sample preparation and instrumentation. Several spectroscopic techniques have been used to study the penetration of the molecules because of the high levels of accuracy and the quick response time of these techniques. Moreover, the samples are not damaged during the investigation.

ER staining levels affect HER2 staining and heterogeneity.

This study explored the relationship of ER expression levels with HER2 staining properties and heterogeneity and discussed the differences in HER2 assessment caused by the 2018 ASCO/CAP guideline updates from that of the 2013 version. HER2-positive breast cancer was divided into three groups of the high hormone receptor expression (LH-high) group, low expression (LH-low) group, or negative (NLH) group to (1) compare differences in the percentage of the HER2 IHC test score of 2 + based on the 2013 ASCO/CAP guideline and in the intratumor heterogeneity of HER2 expression for breast cancer with an IHC score of 3 + among these groups, (2) compare the HER2/CEP17 ratio and the average HER2 copy number, and classified ISH groupings according to the 2018 ASCO/CAP guideline algorithm. (1) Of 244 HER2-positive breast cancers, the cases with a HER2 IHC score of 2 + (n = 54, 22.1%) were significantly more common in the LH-high group (n = 45, P < 0.001). The frequency of heterogeneity was low (n = 25, 10.2%) for the HER2 score of 3 + (n = 190, 77.9%), and significantly higher in the LH-high group (n = 19, 76%, P = 0.002). (2) In a HER2 IHC score of 2 + , Group 2 which is deemed HER2 negative according to the revised 2018 ASCO/CAP guideline was observed in 17 (39.5%) out of 43 cases, of which 16 cases (94.1%) were in the LH-high group. The LH-high group is a heterogeneous group largely consisting of heterogeneous cases with HER2 IHC scores of 2 + or 3 + . NLH, in contrast, is a homogenous group.

A palette of fluorophores that are differentially accumulated by wild-type and mutant strains of Escherichia coli: surrogate ligands for profiling bacterial membrane transporters.

Our previous work demonstrated that two commonly used fluorescent dyes that were accumulated by wild-type Escherichia coli MG1655 were differentially transported in single-gene knockout strains, and also that they might be used as surrogates in flow cytometric transporter assays. We summarize the desirable properties of such stains, and here survey 143 candidate dyes. We eventually triage them (on the basis of signal, accumulation levels and cost) to a palette of 39 commercially available and affordable fluorophores that are accumulated significantly by wild-type cells of the ‘Keio’ strain BW25113, as measured flow cytometrically. Cheminformatic analyses indicate both their similarities and their (much more considerable) structural differences. We describe the effects of pH and of the efflux pump inhibitor chlorpromazine on the accumulation of the dyes. Even the ‘wild-type’ MG1655 and BW25113 strains can differ significantly in their ability to take up such dyes. We illustrate the highly differential uptake of our dyes into strains with particular lesions in, or overexpressed levels of, three particular transporters or transporter components (yhjV, yihN and tolC). The relatively small collection of dyes described offers a rapid, inexpensive, convenient and informative approach to the assessment of microbial physiology and phenotyping of membrane transporter function.

Intraoperative efficacy and clinical outcomes of two commercial staining solutions used in idiopathic epiretinal membrane surgery.

To compare two commercially available staining solutions (MembraneBlue Dual® by D.O.R.C., Netherlands, and TWIN by AL.CHI.MI.A. S.R.L., Italy), in terms of intraoperative handling, staining efficacy and safety, in eyes undergoing surgery for idiopathic epiretinal membrane (ERM). In this observational cross-sectional study, the performance of the dyes used during the procedure (cohesion, ERM and internal limiting membrane [ILM] staining efficacy) was scored by the surgeon using a customized questionnaire after 10 procedures with each of the two dyes. Best-corrected visual acuity (BCVA), central foveal thickness (CFT), blue-light fundus autofluorescence (BAF), and microperimetry-determined retinal sensitivity were reviewed preoperatively and then at 1 and 3months after surgery. ILM staining efficacy with TWIN was scored 2.89 ± 0.33 by the surgeons, which turned out to be higher than with MembraneBlue Dual® (1.90 ± 0.31, P = 0.0002). The cohesion score was 2.70 ± 0.48 for TWIN and resulted significantly higher than with MembraneBlue Dual® (1.60 ± 0.51, P = 0.0010). BCVA, CFT and retinal sensitivity were similar in the two groups, 1 and 3months postoperatively (P nonsignificant for all). Both TWIN and MembraneBlue Dual® dyes showed suitable staining properties and equivalent safety and efficacy profiles, both intra- and postoperatively. The TWIN dye might offer a solution for surgeons who prefer a more cohesive and stable dye.

Ghost cell.

Ghost cells (GCs) have been a curious topic since a great deal of time. Extensive research has been done to deduce the true characteristics and formation of these cells. GCs are balloon-shaped, elliptical, pale eosinophilic epithelial cells with pyknotic nuclei, leaving only a faint outline. In routine H and E staining, these cells give shadowy appearance and hence are also called shadow cells or translucent cells. The present article is an attempt to describe in detail about the origin, microscopic appearance, staining property, immunohistochemistry profile and diagnostic importance of GCs.

Natural stain (Kumkum) formulated by the extract of Curcuma aromatica and slaked lime in histostaining of oral tissues: An observational study.

Background:The pharmacological actions of Curcuma aromatica (wild turmeric) such as anti-inflammatory, antitumor, antifungal, antimicrobial and wound healing have been recognized since ages. However, its role as a natural histological stain has not been explored till date.Aim:To evaluate the efficacy of natural substance-Kumkum prepared from the extract of C. aromatica and slaked lime in staining the biopsied oral tissuesMaterials and Methods:A cohort study that used 60 formalin fixed paraffin embedded soft and hard tissue specimens from institutional archives were subjected to sectioning and stained using Kumkum and hematoxylin and eosin (H and E). The slides were evaluated for their staining efficacy and results were statistically analyzed using Wilcoxon signed-rank test and Independent 't' test.Results:The mean of the overall parameters assessed for staining efficacy did not show statistically significant difference between the study groups in normal and pathological specimens for tooth (P = 0.410 and 0.484), bone (P = 0.133 and 0.157) and soft tissues (P = 0.186 and 0.113), respectively. This suggests that Kumkum staining efficacy is equivalent to that of routine H and E for oral tissues. Structures such as dentinoenamel junction, dentinal tubules, incremental lines of cementum, reversal and resting lines, osteocytic canaliculi, mature and immature bone could be appreciated better in Kumkum stained slides, thereby rendering a special staining property to Kumkum stain.Conclusion:To our knowledge, this study is the first of its kind to have used Kumkum stain obtained from C. aromatica for the differentiation of the components of tooth, bone and soft tissue structures in histostaining of oral tissues. The naturally prepared Kumkum stain possesses dual staining property both in routine and differential staining. This facilitates diagnosis of fibro-osseous lesions, bony, collagen and muscular pathologies. The natural stain also finds application in forensic odontology for age estimation.

Modelowe badania mikrobiomu świń według koncepcji wspólnego zdrowia „One Health” ludzi i zwierząt

Streszczenie Mikrobiom człowieka pod względem liczebności bakterii przewyższa liczbę komórek ludzkiego organizmu. Określany jest jako dodatkowy, „zapomniany narząd” i odgrywa podstawową rolę w utrzymaniu wysokiego statusu zdrowotnego, co jest uwarunkowane zachowaniem pożądanych proporcji i naturalnych relacji między bakteriami a komórkami organizmu gospodarza. Nowe metody diagnostyczne umożliwiają profilowanie nie tylko mikrobiomu człowieka, ale i zwierząt gospodarskich. Coraz szersze zastosowanie w badaniach mikrobiomu ma innowacyjna metoda analityczna, jaką jest sekwencjonowanie nowej generacji NGS (next generation sequencig). Wiele bakterii określa się jako „niehodowalne” lub „niemożliwe do wyhodowania”, metagenomika odegrała istotną rolę w poznaniu tych bakterii, a także przyczyniła się do opracowania nowych pożywek, umożliwiających ich hodowlę. Głównym zastosowaniem NGS w mikrobiologii jest zastąpienie konwencjonalnej charakterystyki patogenów, opartej o ocenę morfologii, właściwości barwienia i cech metabolicznych, ich opisem związanym z genomem. Istnieje kilka platform, tj. „narzędzi diagnostycznych” wykorzystujących zróżnicowane technologie sekwencjonowania DNA m.in. Ion Torrent Personal Genome Machine (PGM), Pacific Biosciences (PacBio) oraz Illumina MiSeq. Badania mikrobiomu trzody chlewnej z wykorzystaniem nowoczesnych technologii sekwencjonowania wydają się więc szczególnie istotne w związku ze zbliżającymi się nieuchronnie zmianami w postępowaniu profilaktycznym i terapeutycznym u zwierząt. Analizy tego typu umożliwiają wnikliwą ocenę wpływu określonych czynników na populacje drobnoustrojów jelitowych oraz poznanie, jak „kształtować” skład mikrobiomu w celu poprawy jakości chowu i utrzymania prawidłowego statusu zdrowotnego świń, wpisując się w koncepcję wspólnego zdrowia ludzi i zwierząt.

Karyotypic diversity among eight Zea mays L. varieties

Staining property of interphase nuclei and prophase chromosomes, diploid chromosome number, total chromosome length (TCL), symmetric and asymmetric indices of karyotype were studied in eight maize varieties released by BARI. 2n = 20 chromosomes were found in Barnali, Mohor, Khoi Vhutta, BS-1, B-5 and BM-7 whereas 2n = 22 chromosomes in China and 2n = 24 chromosomes in B-73 were observed. TCL was highest in Mohor (190.49 ± 5.61 μm) and lowest in B-73 (69.30 ± 2.51 μm). These varieties showed significant variation in cytogenetical parameters. Results obtained are expected to supplement genetic identification of maize varieties in variety conservation efforts.

Efficacy of toluidine blue staining in cervicovaginal cytology over conventional papanicolaou stain

Introduction: Toluidine blue staining (TBS) is a practical, rapid, inexpensive and effective adjunct diagnostic tool. TBS method has been extensively used as a vital stain with metachromatic property for mucosal lesions and exfoliative cytology. Aim and Objectives: To see the efficacy of aqueous Toluidine Blue (TB) stained smears in comparison to conventional smears stained with Papanicolaou (Pap) stain of cervicovaginal smears and to reduce the reporting time of smears and also cutting down on the cost. Materials and Methods: This is a prospective cross sectional study on 240 Cervicovaginal smears received in the Dept. of Pathology, ACPM Medical College College, Dhule over a period of 4 months from September to December 2016. All the satisfactory smears as per Bethesda system of reporting were included in the study. The unsatisfactory smears were excluded from the study. Multiple smears from the symptomatic patients were advised. The conventional Pap stain and Aqueous TBS (1%) was performed on the smears received. Cytomorphology of the smears were studied and compared in reference to staining, timing and efficiency of stains. Results: The efficacy of TBS is equally good as conventional Pap staining. The timing is curtailed from average 30 minutes in Pap staining to 3 minutes in TB staining method. The cost per test was also decreased substantially in TB Staining method. The nuclear staining property by TBS is equally good as in Pap staining most of the times, for rapid diagnosis of lesion. Conclusion: The staining properties of TB Stain are similar and equally good as Pap staining method with reduced turnaround time and are cost effective. Hence TB staining method is beneficial for mass screening of cervicovaginal smears in the camps in Indian scenario. Keywords: Toluidine blue, Pap smears, Cervicovaginal smears, Staining.

Characterization of Morphologic and Cytochemical Staining Properties of Exogenous Materials Mimicking Fungal Organisms Encountered in Skin Biopsies.

Exogenous materials may be encountered in skin biopsies as contamination. Contamination may occur during the biopsy procedure in the clinic or during tissue processing in the laboratory. Although the experienced pathologist may often instinctively ignore clear examples of contamination, such tissue may be a source of confusion for young pathologists and those in training. Foreign materials can typically be recognized as exogenous by morphology, polarizability, and the presence or absence of a tissue reaction, but we have rarely encountered situations in which exogenous materials could be misinterpreted as organisms, either by their morphologic appearance or staining properties. Exogenous materials used during skin biopsy and grossing were embedded in a nonhuman tissue scaffold and prepared into histologic slides. Hematoxylin and eosin (H&E), periodic acid-Schiff with diastase, and Grocott methenamine silver stains were performed, and each material was evaluated under polarized microscopy. Exogenous materials were divided into the following 3 categories with shared morphologic appearances and staining properties: suture materials, plant-based materials, and synthetic materials. We present a comprehensive characterization of the morphologic and cytochemical staining properties of multiple exogenous materials that may contaminate a skin biopsy. This characterization will aid the pathologist by providing a mechanism to identify potential contaminants in skin biopsies.