Periodic acid-Schiff Technique Research Articles

Increased mucous cell population and modulation of Bax/Bcl-2 factors characterize in vivo gastroprotective activity of Cissampelos owariensis in rats

The physiological integrity of the gastric mucosa is dependent on the balance between the mucosal protective and aggressive factors. Medicinal plants or their derivatives generally exhibit gastroprotective effect by promoting the protective factors against the aggressive factors of the gastric mucosa. The study was conducted to elucidate the gastroprotective mechanism of the methanol extract of C. owariensis (MECo) in rats. Twenty male Wistar rats were divided into four groups, which include control groups A and B – given distilled water – and treated groups C and D – animals given 100 and 300 mg/kg MECo respectively for 28 days. After the treatment period, gastric mucosal injury was induced for groups B-D by pyloric ligation method. The gastric tissue of animals was collected, processed for histology (haematoxylin and eosin technique), histochemistry (periodic acid-schiff technique) and immunohistochemical staining (for Bcl-2 & Bax proteins). The results of the gastric histomorphology showed prominent and widespread mucosal erosion in positive control group B compared to normal control group A, while treated groups C and D showed only mild or focal mucosal erosion. Furthermore, the histochemical results showed significant increase in mucous cell population in treated groups C and D compared to positive control group B. The immunostaining results showed significant up-regulation of anti-apoptotic Bcl-2 protein and down-regulation of pro-apoptotic Bax protein in the treated groups C and D compared to the control groups A and B. In conclusion, the findings of this study indicate that the increased mucous cell population and modulation of apoptotic signaling highlights the mechanism of gastroprotective activity of MECo.

Morphogenesis of the ostrich (Struthio camelus) trachea and lung in different embryonic and fetal stages.

The ostrich (Struthio camelus) is an important wild species highlighted in national and international livestock industry. This research was conducted to analyze the development of the ostrich respiratory system during fetal and embryonic stages. A total of 50 fertile ostrich eggs were collected from commercial farms and then incubated at 36.00 - 37.00 ˚C and 25.00 ± 2.00% humidity for 40 days. Sections were taken on days 13, 22, 26, 30, 36, and 42 of incubation from the lung and the cranial, middle, and caudal parts of the neck after decapitation of ostrich embryos and blood drainage. After fixation, processing, blocking, and sectioning, all samples were stained by Hematoxylin and Eosin, Alcian Blue (AB), Van Gieson, and Periodic acid-Schiff (PAS) techniques. It was concluded that the trachea in the 13-day-old embryo and goblet cells (PAS-positive and AB-positive) had incomplete rings of hyaline cartilage and differentiation of mesenchymal to the loose connective tissue. The bronchial stage of the lung was observed in the 22-day-old embryo, pseudoglandular stage in the 26-day-old embryo, and parabrachial and air capillary stage in the 30-day-old embryo. The information obtained from this study will be useful for diagnosing pathologies affecting this vital system and results in improving industrial breeding management.

Evaluation of Protective Effect of Ajwa Seed And Fruit on Renal Histopathological Changes in Diabetic Nephropathic Rats

Objective: To investigate the protective effect of Ajwa date seed and fruit on renal histological changes in alloxan induced diabetic rats. Methods: This was an experimental study and was conducted in Post Graduate Medical Institute. The duration of study was 6 weeks. In this study random allotment of 32 rats was done in four groups. Group 1 was treated as control. Diabetes was induced in the 2nd, 3rd and 4th group by alloxan injection intra peritoneally. Group 2 was diabetic non treated while group 3 and 4 were treated with Ajwa seed and flesh respectively. After six weeks, animals were anaesthetized and kidneys were then removed without delay and weighed. Kidney paraffin sections were prepared and stained with hematoxylin& eosin (H&E) and with Periodic acid Schiff (PAS) technique. Glomerular diameters were estimated. Glomerular volume determined by stage micrometer. Vascular, tubular injury and glomerular sclerosis were studied semi quantitatively. Results: The data showed that Ajwa date seed significantly reduced hyperglycemia but did not normalize the fasting blood glucose. We found exceedingly significant improvement in kidney weight, glomerular diameter, tubular and vascular injury with Ajwa date seed suggesting reduction in diabetic nephropathy. Ajwa seed diet found more effective in reducing nephropathy than Ajwa fruit diet. Current study displayed that the seed of Ajwa showed significant improvement in renal histological characteristics in diabetic rats. Conclusion: The findings showed that Ajwa date seed and flesh reduce loss of tubular and vascular damage in alloxan induced diabetes. Key Words: Oxidative stress, Kidney, Diabetes, Ajwa, Antioxidant, histopathology How to cite: Imran I., Mansoor M., Naqvi F., Akhtar M., Arshad W., Khan F. Evaluation of protective effect of Ajwa seed and fruit on renal histopathological changes in diabetic nephropathic rats. Esculapio 2021;17(01):104-109

Impact of acute fonofos exposure on skeletal muscle of zebrafish: Histopathological and biometric analyses

It is widely known that pesticides generally do not show target specificity, and off-target species are strikingly affected by these chemicals. In the current work, histological changes in skeletal muscles of zebrafish (Danio rerio) caused by fonofos, an acetylcholinesterase (AChE) inhibitor organophosphate insecticide, were examined. Zebrafish were treated with 1 mg/L, 2 mg/L and 4 mg/L of fonofos for 96 hours. Skeletal muscle samples were removed from the pectoral region and embedded in paraffin. Sections were stained with Mayer's Hematoxylin and Eosin, Gomori's Trichrome and Periodic Acid Schiff techniques. Histopathological alterations were investigated by light microscopy. Fibrosis, intramyofibrillar vacuoles, disintegrated myofibrils, splitting of myofibers, atrophic and disappeared fibers, histoarchitectural loss, necrosis and progressive decrement in glycogen content were noted. Muscle fiber diameter measurements were also performed. Statistical analysis showed that measured fiber diameters of all fonofos exposed groups were significantly different from the control group, and they decreased in a concentration-dependent manner. These results suggested that fonofos caused significant myoarchitectural impairments in non-target freshwater zebrafish.

Transcriptomic and microstructural analyses in Liriodendron tulipifera Linn. reveal candidate genes involved in nectary development and nectar secretion

BackgroundNectar is a major floral attractant and reward for insects that ensures pollination. Liriodendron, a genus of the Magnoliaceae family, includes only two relict species, L. chinense and L. tulipifera, which are considered “basal angiosperms” according to plant evolutionary history. The flowers of Liriodendron plants are insect pollinated and secrete nectar to attract pollinators. To date, the morphology and anatomy of nectaries, the mechanism of nectar secretion and the molecular mechanism of nectary development in Liriodendron remain poorly understood.MethodsIn this study, we examined the nectary surface cells and change in starch in L. tulipifera by using scanning electron microscopy and periodic acid-Schiff techniques to select appropriate samples for subsequent research. Transcriptome sequencing was of the top and middle parts of immature nectaries and the middle part of mature and postsecretory nectaries in L. tulipifera was performed. We evaluated the expression profiles of 21 DEGs that are closely related to nectary development and nectar secretion for real-time quantitative PCR analysis.ResultsL. tulipifera nectaries are starch-storing nectaries and are located in the top and middle parts of L. tulipifera petals. After analyzing the RNA-seq data, we obtained 115.26 Gb of clean data in 12 libraries and mapped the results to the L. chinense reference genome with 71.02–79.77% efficiency. In total, 26,955 DEGs were identified by performing six pairwise comparisons. The flavonoid biosynthesis, phenylpropanoid biosynthesis, anthocyanin biosynthesis and starch and sucrose metabolism pathways were enriched and related to nectar secretion and pigment change. We identified 56 transcription factor families, and members of the TCP, Trihelix, C2H2, ERF, and MADS families changed dynamically during nectary development. Moreover, to further verify the accuracy of the RNA-seq results, we validated the expression profiles of 21 candidate genes.ConclusionsWe evaluated the nectary development and secretion processes comprehensively and identified many related candidate genes in L. tulipifera. These findings suggest that nectaries play important roles in flavonoid synthesis and petal color presentation.

Hystometric evaluation of nickel chronic exposure effects on large instestine of adult Wistar male rats

The ingestion of considerable amounts of water or food contaminated with nickel can be very toxic. The present work was conducted aiming to evaluate the effects of nickel exposures on ascending colon of adult Wistar male rats at hystometric level. We used 12 animals that were divided in a control (ingested uncontaminated water) and a nickel-contaminated (i.e., 25 mg de nickel/L of water) groups. Nickel chloride was offered in declorinated water and the experiment had a 56 days exposure period. A portion of the ascending colon was removed of the animals and subjected to hystological labelling processes using blue toluidin (for general hystometric description), Alcian Blue (AB, for acid mucins) and periodic acid-Schiff (PAS) technique (for neutral mucins). The potential differences between groups were desgined by applying the Whitney test and t test (p < 0.05). The crypts were smaller for the nickel-contaminated group, even though these organism exhibited broader and higher crypts. Nickel-contaminated animals exhibited a smaller amount of calyceform cells with AB and PAS positive reactions as well as a less mucus quantities when compared with nickel-uncontaminated animals. Such reductions on the amount of calyceform cells with AB and PAS positive reactions may be related wiht the shallower crypts, which possibly reduced the synthesis and secretion of mucins, compromissing the functional aspects (e.g., lubrification and intestinal mucosa protection) of the nickel-contaminated large intestines. Interestingly, the wider and higher crypts and higher epithelium collumn on the nickel-contaminated animals may represent a relevant trade-off for the intestinal mucosa protection.

Mucin histochemical profile in a variety of human colonic non-neoplastic and neoplastic lesions

Mucin histochemical profile in a variety of human colonic non-neoplastic and neoplastic lesions - JDPO- Print ISSN No: - 2581-3714 Online ISSN No:- 2581-3706 Article DOI No:- 10.18231/j.jdpo.2019.034, IP Journal of Diagnostic Pathology and Oncology-IP J Diagn Pathol Oncol

The Use of One Normal Hydrochloric Acid as a Background Cleaner for Periodic Acid–Schiff Staining Technique

Abstract Objectives Sulfurous rinsing was considered nonessential as contained in the published periodic acid–Schiff technique because it did not clear the background better than tap water washing. The objective of this work is to elucidate the use of 1N HCl as a better background cleaner in the periodic acid–Schiff staining technique. Methods This is an experimental research. Normal samples of antrium, kidney, spleen, muscle, liver, and appendix, as well as histopathologically diagnosed samples of glycogen storage disease of the liver, alpha-1-antitrypsin deficient liver, aspergillosis nasal cavity sample, and glomerulosclerosis samples were obtained from the pathological archive. These samples were stained with a newly purchased commercially prepared Schiff reagent and overstained commercially prepared Schiff’s solution causing nonspecific background staining and laboratory-prepared Schiff reagent. Oxidation was done with 1% periodic acid for 5 minutes, washed for 5 minutes with distilled water, and stained for 10 minutes in Schiff’s reagent, but 20-minute staining was observed for the overstained Schiff reagent, oxidized in running tap water for 15 minutes. It was then rinsed in 1N HCl for 20 seconds, washed in tap water for 2 minutes, and counterstained with hematoxylin for 2 minutes. The same procedure was done for another set of slides as standards without 1N HCl rinsing done. The slides were examined under the microscope and compared for specificity. Results Slides rinsed in 1N HCl showed a clear differentiation of strong, weak, and non-PAS-positive substances. The overall effect of these staining characteristics was more pronounced in the use of laboratory-prepared Schiff. Conclusion The use of 1N HCl as a differentiator made the PAS technique more specific in its staining characteristics, especially with the use of laboratory-prepared Schiff’s reagent, which is commonly used in developing countries owing to a high cost of commercially prepared Schiff’s solution.

Histological Effects of Sofosbuvir on The Kidney of Male Albino Rats and The Possible Protective Role of Vitamin C

Aim of the work: hepatitis C virus (HCV) infection is a major worldwide public health issue. Direct Acting Antiviral Drugs (DAA) is a revolution that occurred in treatment of hepatitis C virus (HCV). The Second class of DAA is HCV NS5B polymerase inhibitors (Which included Sofosbuvir) was shown to be effective in suppression of HCV replication and became an important component of currently recommended regimens. Material and Methods: forty adult male albino rats (Rattus norvigicus) weighting 140±20g were selected for this 36-days experiment The rats were randomly categorized into six groups (10 animals each for the first four groups and the remaining groups VV periodic Acid Schiff technique (PAS) for studying of general carbohydrate ; Masson’s trichrome stain for detecting the collagen fibers and transmission electron microscopy (TEM) to detect the ultrastructural changes induced by Sofosbuvir. Results: the present study showed that group II which received the therapeutic dose of Sofosbuvir (0.06 mg/gm./body weight) showed renal tubular affection compared to the control rats. Tubular affection included massive tubular vacuolization especially cells of the proximal and distal convoluted tubules. Group III which received the toxic dose of Sofosbuvir (0.25 mg/gm./body weight) resulted in glomerular affection compared to the control rats. Conclusion: the present study provided promising results about reduction of Sofosbuvir nephrotoxicity by the concomitant use of vitamin C.

Von Hansemann Cells from Fresh Urine Sediment Samples in the Diagnosis of Malakoplakia.

Background:Malakoplakia is characterized by the presence of plaques with macrophages containing inclusion bodies. The diagnosis of this disease is carried out by biopsy of the lesion. The objective of this paper was to assess the value of fresh urine sediment in the diagnosis of malakoplakia.Materials and Methods:Five suspected cases of malakoplakia that showed macrophages with inclusions called bodies of Michaelis-Gutmann (von Hansemann cells) in unstained urine sediment were processed with Papanicolaou, Giemsa, and periodic acid-Schiff (PAS) stains. Four of the five patients had a history of cystitis and had developed antibiotic resistance. The other patient had the characteristics cells in a routine urinalysis.Results:Papanicolaou stain revealed intracytoplasmic eosinophilic or basophilic bodies, single or multiple in macrophages. Such bodies were stained deep red with PAS technique. Giemsa stain showed these bodies with a faint basophilic coloration, sometimes with a central core. Bladder biopsies established the definitive diagnosis, showing bodies within and outside macrophages, with a concentric “birds-eye” or “owl-eye” (targetoid) appearance.Conclusions:Finding of von Hansemann cells in fresh urine sediment of patients with cystitis and a history of resistance to antibiotic scan leads to the diagnosis of malakoplakia. Giemsa stain can show in some cases the characteristic central core of Michaelis-Gutmann bodies. Malakoplakia is probably the result of an acquired defect in macrophage function causing impairment of bactericidal activity. A correct diagnosis is important because the spread to ureters with bilateral stenosis and obstruction can lead to kidney failure.

Identification of Candida species in type 2 diabetic patients using three different staining techniques

Oral candidiasis is a condition commonly seen in patients with systemic diseases such as diabetes, a common and globally growing health problem associated with many risk factors. The objective of the study was to identify Candida albicans in cultures and compare various cytologic stains used for diagnosis of oral candidiasis. Ninety type 2 diabetic individuals were chosen for the study group. The control group contained 30 non-diabetic individuals. Random blood sugar levels were analyzed for all diabetic and non-diabetic patients. Gram stain, periodic acid-Schiff technique (PAS) and calcofluor white were used for staining buccal mucosa smears collected from all individuals. An oral rinse was collected from all subjects and inoculated on Sabouraud dextrose agar (SBA) to confirm Candida positivity. Results for Candida positivity from one patient show variability between the three stains. Cultures showed higher values of these variables i.e. specificity, sensitivity and positive predictive values for Candida identification as compared to the staining results. This study showed detection of C. albicans in the oral cavity was better using SBA culture media than stained exfoliative smears.

Morpho-histochemical characterization of the salivary glands of semi-engorged Amblyomma triste (Koch, 1844) (Acari: Ixodidae) female ticks

This study presents the morphological and physiological characterization of the salivary glands of semi-engorged Amblyomma triste females. Unfed individuals were placed on New Zealand White rabbits for feeding and the females, after 4 days, were collected, dissected and the salivary glands were submitted to the application of histological (hematoxylin–eosin technique) and histochemical tests for the detection of protein (bromophenol blue technique, polysaccharides (periodic acid–Schiff technique), lipid (Nile blue technique) and calcium (von Kossa technique). The histological results show that the glandular tissue is composed by a system of ducts and three types of acini (I, II and III). The acini I are formed by a large central cell surrounded by several smaller agranular peripheral cells. Acini II are formed by cells a, c1, c2, c3 and c5, which are full of secretion granules. Acini III are constituted by cells d, e and f; the former two contain secretion granules, the latter is agranular. The glandular histochemical composition was also verified. Data obtained here will certainly help in the understanding of the cellular morphology and of the general physiology of these organs in this specie, providing important information for the creation of scientific bases which will contribute for the development of more specific and efficient methods of control.

Polysaccharides as Constituents in Chromosome Organization: A Study on Meiotic Chromosomes of Grasshopper and Polytene Chromosomes of Dipterous Flies

Study on meiotic chromosomes of grasshopper, Gesonula punctifrons and interphase polytene chromosomes from Dipteran larvae as of Chironomus striatipennis and Drosophila melanogaster following staining by periodic acid-Schiff technique revealed that chromosomes contained polysaccharides as an integral part of their organization. PAS +ve nature of the chromosomes both at highly condensed state as available during meiotic cell division and at extended state as in polytene chromosomes supports the idea that chromosomes contain polysaccharides as one of the constituent biological macromolecules. PAS +ve chromosomes appeared to be fluorescent under fluorescence microscope and fluorescence was found to be more or less uniform along the whole length of the meiotic chromosomes, while in case of polytene chromosomes intense fluorescence could be noticed along the band regions of the chromosomes.

Cytochemical study of developing anthers of Amomum villosum Lour.

We analyzed anther development in Amomum villosum Lour. (Zingiberaceae) using the periodic acid-Schiff's technique and Sudan black staining to test for the presence of starch and lipids, respectively. Our analyses showed that microspore mother cells of A. villosum lack typical callose walls, and numerous lipid granules appear in the cells early in development. Some starch granules are present in anther wall cells, but not in tapetal cells. After meiosis, numerous lipid granules remain unchanged in the microspores. During microspore development, some small starch granules first appear in the central cell region, and then the starch granules increase in size. After microspore division, the bicellular pollen grains become filled with starch and lipids, and remain in this state until the pollen grains reach maturity. At anthesis, the anther wall of A. villosum consists of several layers of endothecium cells with an evidently thickened radial wall, and some layers of parenchyma cells containing numerous starch granules.

Parahippocampal corpora amylacea and neuronal lipofuscin in human aging

Abstract The aim of this research was to quantify the number of corpora amylacea and lipofuscin-bearing neurons in the parahippocampal region of the brain. Right parahippocampal gyrus specimens of 30 cadavers were used as material for histological and morphometric analyses. A combined Alcian Blue and Periodic Acid-Schiff technique was used for identification and quantification of corpora amylacea and lipofuscin-bearing neurons. Immunohistochemistry was performed using S100 polyclonal, neuron-specific enolase and glial fibrillary acidic protein monoclonal antibodies for differentiation of corpora amylacea and other spherical inclusions of the aging brain. Cluster analysis of obtained data showed the presence of three age groups (median age: I = 41.5, II = 68, III = 71.5). The second group was characterized by a significantly higher numerical density of subcortical corpora amylacea and number of lipofuscin-bearing neurons than other two groups. Values of the latter cited parameters in the third group were insignificantly higher than the first younger group. Linear regression showed that number of parahippocampal lipofuscin-bearing neurons significantly predicts numerical density of subcortical corpora amylacea. The above results suggest that more numerous parahippocampal region corpora amylacea and lipofuscin-bearing neurons in some older cases might represent signs of its’ neurons quantitatively-altered metabolism.

Evaluation of immunohistochemistry for the diagnosis of sporotrichosis in dogs

The aim of this study was to apply immunohistochemistry (IHC) for the diagnosis of canine sporotrichosis and to compare this method with the Grocott's silver stain (GSS) and periodic acid Schiff (PAS) techniques. Eighty-seven dogs with sporotrichosis (group 1) and 35 with American tegumentary leishmaniosis (ATL) (group 2) were studied. The fungus was detected in group 1 by GSS, PAS and IHC. IHC was also applied to group 2 to evaluate the occurrence of cross-reactions. PAS, GSS and IHC detected yeast cells in 19.5%, 43.7% and 65.5% of the group 1 cases, respectively. The detection of intracellular antigens of Sporothrix schenckii by IHC increased the sensitivity of the histological diagnosis to 80.5%. No positive reaction was observed in ATL lesions. The results suggest that IHC may be indicated for the diagnosis of sporotrichosis because of its higher diagnostic sensitivity.

Ameliorative Effects of Ethanolic Leaf Extract ofAzadirachta indicaon Renal Histologic Alterations in Streptozotocin-Induced Diabetic Rats

We studied the effect of ethanolic leaf extract of Azadirachta indica (AIE) on the microanatomy of the kidney of streptozotocin-induced diabetic rats. Thirty male Wistar rats (161-190 g) were randomly assigned to one of five treatment groups of six animals each: control, diabetic, diabetic + AIE, diabetic + metformin, AIE only. Diabetes was induced with a single intraperitoneal dose of streptozotocin (70 mg/kg body weight). AIE and metformin were administered orally for 50 days (50 d) at 500 mg/kg bw/d and 350 mg/kg bw/d, respectively. Blood glucose was estimated by glucose oxidase method; plasma urea and creatinine were assayed; and paraffin sections of the kidney were stained by periodic acid-Schiff technique. Untreated diabetic rats exhibited marked hyperglycemia. Renal histopathology of these animals showed features of diabetic nephropathy, with nodular glomerulosclerosis and vacuolation of proximal tubule cells (Armanni-Ebstein phenomenon). These feature were absent in the diabetic rats treated with AIE. Besides, plasma urea and creatinine were not significantly different from the control in this group (p > 0.05), in contrast to the untreated diabetic rats, where significant increases in these markers (p < 0.05). These findings showed that the leaf extract of Azadirachta indica ameliorates hyperglycemia and diabetic nephropathy in rats.

Light, Scanning and Transmission Electron Microscopical Study on the Oviduct of the Ostrich (Struthio camelus)

Nine mature female ostriches (Struthio camelus) aged 37- 49 months were used for the present study. The samples were taken during the period from September to November from Almasreya ostrich farm, immediately after sacrification and evisceration. Pieces from the different segments of the left oviduct were taken, then fixed in 10% neutral buffered formalin and Bouin’s and Susa solutions for the light microscopic study and in 2 % gluteraldehyde for both scanning and transmission E/M examination. For light microscopic study the slides were stained with Harri's hematoxylen and eosin, Crossman’s trichrome stain, Weigert's elastic tissue stain and Periodic acid Schiff technique (PAS).The right oviduct is rudimentary, while the left one is well developed and is formed of five segments, namely: infundibulum, magnum, isthmus, uterus, and vagina. The scanning E/M revealed that the lining epithelium of the oviduct is folded and formed of pseudostratified columnar ciliated cells. The cilia were slender, dense uniformly arranged, and identical in length. In magnum, isthmus and uterus the scanning E/M study revealed the presence of openings between the cilia for the secretion of the tubular glands of the lamina propria.

Structural study of the salivary glands of Anocentor nitens (Acari: Ixodidae) during the feeding cycle

The salivary glands of Anocentor nitens (Neumann,1897) occur in pairs and are located in the anterolateral region of the general cavity, with milky white color and approximately equal sizes. They consist of a secretory portion and an excretion duct. In some glandular acini, all the cells had a basophilic appearance they were stained by hematoxylin, whereas others presented cells with different staining affinities. In this work, we describe the variations observed in these glands during the feeding cycle of ticks [after feeding (0 h) and successively at 24, 48, 72, 96, 120, and 144 h]. The cells stained by hematoxylin were shown to be more reactive to Alcian blue, thus demonstrating the presence of acid glycosaminoglycans, whereas those stained using eosin presented weak or no reaction. A strong reaction was found by the use of the periodic acid-Schiff (PAS) technique, thereby suggesting the presence of glycogen and/or glycoconjugates containing hexose, confirmed by using salivary amylase before PAS, with partial destaining of the slides. Continuing presence of residual staining in these cells suggests the presence of glycoconjugates containing hexose. Cells with nuclei of circular outline and few granules (of different sizes) were found in type II acini, 72 h after collection. Type I acini presented wide lumina and walls composed of larger numbers of cells of cubic to cylindrical shape. The pronounced degranulation shown in this study over the course of the feeding cycle was associated with the release of substances for oviposition.

Potential Use of a Monoclonal Antibody for the Detection of Candida Antigens in an Experimental Systemic Candidiasis Model

The Candida species are the most common fungal pathogens of systemic candidiasis. The diagnosis of invasive candidiasis remains a laboratory and clinical challenge. Thus, development of diagnostic assays to detect systemic candidiasis and to identify Candida virulence factors and associated pathogenesis through immunohistochemistry using specific monoclonals and polyclonals will be useful. Inbred Balb/c mice were immunized with C. albicans antigens, and blood was checked for the presence of reactive antibodies using ELISA. Fusion was performed using the harvested spleen cells and NS1 myeloma cells, and the clones were screened for the presence of antibody producing hybrid cells by dot-blot. Western blot analysis showed that the L2D10 monoclonal antibody was reactive against the antigens with molecular weight of 20 kDa. Experimental systemic candidiasis in mice was induced through intravenous injection of C. albicans and all the vital organs were collected for immunohistochemistry study. The monoclonal antibody reacted to surface epitopes on the yeast cells, germ tubes, and hyphae, and to immune complexes. It was used with the polyclonal antibody in a sandwich ELISA for the detection of circulating antigens in experimental candiadiasis in mice. Antibody levels were also determined using the ELISA method, and the antibody levels of C. albicans infected mice were increased compared with uninfected animals. The monoclonal antibody was used in immunoperoxidase and immunofluorescence techniques for the detection of fungal infection in tissue sections and was found to be more sensitive than conventional periodic acid Schiff or silver staining techniques. This monoclonal antibody may serve as potential primary capture antibodies for the development of a rapid diagnostic test for human systemic fungal infection.