Stains are applied to impart contrast to the tissue and identify particular features of interest. However, the use of synthetic dyes as staining reagents has been associated with significant human health challenges an...Stains are applied to impart contrast to the tissue and identify particular features of interest. However, the use of synthetic dyes as staining reagents has been associated with significant human health challenges and pollution of the ecosystem. These developments have necessitated a shift towards using natural dyes that are eco-friendlier and readily available. We investigated the staining reaction patterns of teak tree leaves (Tectona grandis) dye extracts and explored their suitability as a cytoplasmic stain in micromorphological assessments. Dye extracts were prepared using acetone, methanol, and ethanol as solvents from air-dried (under shade) teak tree young leaves. The dye extracts were applied as a counterstain and evaluated against eosin in formalin-fixed paraffin-embedded (FFPE) bovine tissue sections at varying concentrations and different staining times. Teak tree leaves (Tectona grandis) dye extracts produced relatively varying staining intensities of reddish-brown cytoplasmic coloration when used on bovine tissue at different concentrations and staining times comparable to eosin and with blue-purple hematoxylin nuclear stain. The present study showed that Tectona grandis leaf dye extracts provide an excellent cytoplasmic staining pattern and can be used as an alternative counterstain in routine H&E staining techniques.展开更多
Background:TTC(2,3,5-triphenyltetrazolium chloride)staining is the most commonly used method in identifying and assessing cerebral infarct volumes in the transient middle cerebral artery occlusion model.Given that mic...Background:TTC(2,3,5-triphenyltetrazolium chloride)staining is the most commonly used method in identifying and assessing cerebral infarct volumes in the transient middle cerebral artery occlusion model.Given that microglia exhibit different morphologies in different regions after ischemic stroke,we demonstrate the superiority and necessity of using TTC-stained brain tissue to analyze the expression of various proteins or genes in different regions based on microglia character.Methods:We compared brain tissue(left for 10 min on ice)from the improved TTC staining method with penumbra from the traditional sampling method.We identified the feasibility and necessity of the improved staining method using real time(RT)-PCR,Western blot,and immunofluorescence analysis.Results:There was no protein and RNA degradation in the TTC-stained brain tissue group.However,the TREM2 specifically expressed on the microglia showed a significant difference between two groups in the penumbra region.Conclusions:TTC-stained brain tissue can be used for molecular biology experiments without any restrictions.In addition,TTC-stained brain tissue shows greater superiority due to its precise positioning.展开更多
Introduction: This study aimed to perform routine seminal fluid analysis, sperm DNA fragmentation, and sperm function tests at the chromatin maturation level and evaluate pregnancy in the patients passing intrauterine...Introduction: This study aimed to perform routine seminal fluid analysis, sperm DNA fragmentation, and sperm function tests at the chromatin maturation level and evaluate pregnancy in the patients passing intrauterine insemination before starting Intrauterine Insemination (IUI) method. Materials and Methods: In this prospective study, 111 couples who underwent Intrauterine Insemination (IUI) in unexplained infertility patients were admitted to Al-Farah IVF and assisted reproductive center in Baghdad, Iraq between November 2020 and February 2021 were evaluated. Semen fluid analysis was performed based on (WHO 4th) guiding rules. In addition, Sperm Chromatin Dispersion (halo test) and sperm maturation were performed with Aniline Blue Stain (ABS). Results: Sperm Chromatin Dispersion (SCD) groups were compared in terms of pregnancy outcome;the positive pregnancy rate was found to be above in the normal SCD groups (p = 0.0005). In addition, Aniline Blue Stain (ABS) groups were compared in the terms of pregnancy outcome;the positive pregnancy rate was found to be higher in the normal ABS group (p = 0.017). Conclusion: Our study showed that the use of DNA fragmentation (SCD) and sperm maturation tests (ABS) together with routine semen analysis in intrauterine insemination cases will make a significant contribution to the prediction of Intrauterine Insemination (IUI) increased results. So, these results indicate a defect in the effect of DNA fragmentation on the outcome of intrauterine insemination.展开更多
Port-wine stain(PWS)is a congenital capillary malformation that occurs in 0.3%–0.5%of newborns.The pulsed dye laser is the current gold standard treatment for PWS;however,its efficacy is poor.Photosensitizer photodyn...Port-wine stain(PWS)is a congenital capillary malformation that occurs in 0.3%–0.5%of newborns.The pulsed dye laser is the current gold standard treatment for PWS;however,its efficacy is poor.Photosensitizer photodynamic therapy(PDT)is considered a promising treatment for PWS.Here we provide a comprehensive overview of PDT.展开更多
In the early days of deciphering the injured neuronal tissues led to the realization that contrast is necessary to discern the parts of the recovering tissues from the damaged ones.Early attempts relied on available(a...In the early days of deciphering the injured neuronal tissues led to the realization that contrast is necessary to discern the parts of the recovering tissues from the damaged ones.Early attempts relied on available(and often naturally occurring)staining substances.Incidentally,the active ingredients of most of them were small molecules.With the advent of time,the knowledge of chemistry helped identify compounds and conditions for staining.The staining reagents were even found to enhance the visibility of the organelles.Silver impregnation identification of Golgi bodies was discovered in owl optic nerve.Staining reagents since the late 1800s were widely used across all disciplines and for nerve tissue and became a key contributor to advancement in nerve-related research.The use of these reagents provided insight into the organization of the neuronal tissues and helped distinguish nerve degeneration from regeneration.The neuronal staining reagents have played a fundamental role in the clinical research facilitating the identification of biological mechanisms underlying eye and neuropsychiatric diseases.We found a lack of systematic description of all staining reagents,whether they had been used historically or currently used.There is a lack of readily available information for optimal staining of different neuronal tissues for a given purpose.We present here a grouping of the reagents based on their target location:(I)the central nervous system(CNS),(II)the peripheral nervous system(PNS),or(III)both.The biochemical reactions of most of the staining reagents is based on acidic or basic pH and specific reaction partners such as organelle or biomolecules that exists within the given tissue type.We present here a summary of the chemical composition,optimal staining condition,use for given neuronal tissue and,where possible,historic usage.Several biomolecules such as lipids and metabolites lack specific antibodies.Despite being non-specific the reagents enhance contrast and provide corroboration about the microenvironment.In future,these reagents in combination with emerging techniques such as imaging mass spectrometry and kinetic histochemistry will validate or expand our understanding of localization of molecules within tissues or cells that are important for ophthalmology and vision science.展开更多
Introduction: Reports indicate that fluorescent staining of smears increases sensitivity of direct microscopy;so ZN staining is being replaced with fluorescent microscopy in RNTCP in India. Chemical processing and spu...Introduction: Reports indicate that fluorescent staining of smears increases sensitivity of direct microscopy;so ZN staining is being replaced with fluorescent microscopy in RNTCP in India. Chemical processing and sputum concentration may also improve sensitivity of microscopy. Objective: To compare the sensitivity and specificity of microscopy for AFB using ZN and fluorescent stains in direct and concentrated specimen with culture as gold standard. Methods: Morning sputum specimen of patients, suspected of having pulmonary tuberculosis, over a period of 6 months was subjected to direct microscopy using fluorescent stain;the same slide was over-stained with ZN stain. Same sputum sample was concentrated by Petroff’s method and subjected to fluorescent microscopy followed by ZN microscopy and finally to culture for AFB. Results: Sensitivity of fluorescent stained concentrated sputum samples was maximum and of ZN stained unprocessed sputum samples was minimum. Specificity of three of the methods was equal at 0.96 but of ZN stained concentrated sputum smears was 0.97. Sensitivity of total fluorescent stains was 0.85 (Specificity 0.96) and sensitivity of total ZN stained smears was 0.80 (Specificity 0.96). Discussion: We used same smear for fluorescent and ZN stains, so smear related variability is decreased. Blinding for microscopy was practically complete. Conclusion: The sensitivity of sputum microscopy for AFB can be increased by concentrating the sputum and using fluorescent microscopy. The specificity remains high in all the methods.展开更多
In order to find a simple and conve-nient method of diagnosis and to probe intothe value of auricular diagnosis in malignanttumors so as to substantiate and
Negative staining is an effective method that can be used for electron microscopic study to observe fine structural morphology without destruction of bacterial structure. Although uranium acetate is used worldwide as ...Negative staining is an effective method that can be used for electron microscopic study to observe fine structural morphology without destruction of bacterial structure. Although uranium acetate is used worldwide as a general dyeing solution, it is extremely difficult to use it by a new purchase at a research institution because it falls under the nuclear regulation substance in Japan. Therefore, we examined alternative reagents for negative staining that could replace uranium acetate through bacterial observation with an electron microscope. Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, and Streptococcus pyogenes were examined by four stain reagents (phosphotungstic acid (PTA), EMstainer, TI blue, and uranium acetate). Pre cultured bacteria were stained with each stain reagents on a copper grid, washed with PBS, and observed with a transmission electron microscope. In the comparison between bacterial structures, the cell wall structure and bacterial flagella could be observed well in the order of PTA, EMstainer, and uranium acetate. With TI blue staining, flagella could be observed very poorly. In comparison between bacteria, gram negative bacteria such as Escherichia coli and Pseudomonas aeruginosa, could be observed well as compared with gram positive cocci such as Staphylococcus aureus and Streptococcus pyogenes. The uranium acetate looked very coarse in background particles. Since crystals tend to precipitate, TI blue also required filtering, and electron beams were absorbed by the agglomerated crystals, and the frequency of electronic burning occurred high frequency. In this study, there was clear difference in the observation conditions depending on the type of bacteria and the kind of the staining reagents. Especially, it was confirmed that good negative staining features of Pseudomonas aeruginosa by electron microscope were obtained by PTA and EMstainer staining. These alternative reagents are considered to be a candidate for a negative staining.展开更多
Objective: To explore the effect of gardenia blue pigment on the anterior capsule staining and its safety performance. Methods: In this study, New Zealand white rabbits were used as the research object to test the eff...Objective: To explore the effect of gardenia blue pigment on the anterior capsule staining and its safety performance. Methods: In this study, New Zealand white rabbits were used as the research object to test the effect of gardenia blue pigment on the anterior capsule staining and anterior segment toxicity. In this study, gardenia blue stains of different concentrations (5%, 4%, 3%, 2%, 1%, 0.5%) were prepared to stain the anterior capsule of rabbit crystals, and the effect of the stain was observed to determine the optimal concentration of the stain. Twenty-seven healthy New Zealand white rabbits without eye disease were randomly divided into three groups: gardenia blue staining group;bio-blue positive drug group;physiological saline group. The intraocular pressure, anterior chamber inflammation, corneal endothelial injury, and anterior chamber angle pathological changes were measured in 1D, 7D, and 14D. Results: The results of this study showed that the effect of gardenia blue pigment on the anterior capsule of the lens was the best when the concentration of blue pigment was 2%. There was no change in intraocular pressure in the anterior chamber for a short period of time. There is no damage to the corneal endothelium. Conclusion: The results of this study show that the effect of gardenia blue pigment on the anterior capsule is best when the concentration of blue pigment is 2%, and the stain has no obvious toxic and side effects on the anterior segment of the eye.展开更多
Stains and staining methods significantly assist in diagnoses in medical research and health care.Certain color of a dye can identify the location of a tumor within a specimen.Applying histochemical-staining enabled m...Stains and staining methods significantly assist in diagnoses in medical research and health care.Certain color of a dye can identify the location of a tumor within a specimen.Applying histochemical-staining enabled morphological identification of fibrin in the lymphoid tissue during cancer progression.Staining methods in combination with the LSFM(light-sheet fluorescence microscopy)allowed tracing the drug penetration,development and spread of tumors.The Curcumin dye is in use for labelling and imaging of Aβplaques in post-mortem brain tissue.Immunofluorescent staining methods are employed in detection of some important proteins in early diagnostic changes relevant to heart damage.The methods are developed in medical research to include stem cells and tissue engineering,cell culturesproperties and capabilities,connective tissues and extracellular matrix,nervous system,musculoskeletal system;respiratory system,liver and gastrointestinal tract,and male and female reproductive systems.展开更多
One of the significant features of The Human Stain is that it lays bare the reality of American society and censures the Anglo-Saxon or white discourse.What are equally important are the multiple narrative strategies ...One of the significant features of The Human Stain is that it lays bare the reality of American society and censures the Anglo-Saxon or white discourse.What are equally important are the multiple narrative strategies that Philip R oth employs in the novel.T his paper focuses mainly on the elaboration of such narrative strategies as mimesis and diegesis,complicated narrative time,shifting focalizations,and narration or non-narrative comments,so as to point out that these narrative strategies add colors to its( postmodern) artistic features,becoming an integral component of this novel.展开更多
Background: Tuberculosis is a highly infectious disease and India has the highest burden with it. Diagnosis of tuberculosis in many countries is still dependent on microscopy. Although its sensitivity is low in compar...Background: Tuberculosis is a highly infectious disease and India has the highest burden with it. Diagnosis of tuberculosis in many countries is still dependent on microscopy. Although its sensitivity is low in comparison to culture and molecular methods, its sensitivity can still be improved by using fluorescence staining method and processing of samples by homogenization and concentration method. Material and methods: Samples were collected from all newly registered suspected cases of tuberculosis in tertiary care hospital from outward and indoor department during a period of one year. Smears were prepared for Ziehl Neelsen stain and fluorescence stain both before and after homogenization and concentration procedure by 4% NAOH-2.9% sodium citrate method and results of them were interpreted according to RNTCP criteria for grading of sputum samples. All the samples were cultured in liquid culture MGIT system (Mycobacterial Growth Indicator Tube) and results of microscopy were compared with liquid culture taken as gold standard. Data were analyzed by using SPSS software version 16. Result: 350 samples were collected during study period. Out of 350 samples, 48 samples were positive for M. tuberculosis by MGIT system. In comparison with MGIT system, sensitivity of Z N stain for detection of acid fast bacilli was 77% before decontamination procedure, which was increased up to 85.42% after decontamination and concentration process. Sensitivity of fluroscence stain was 85.42% before processing, which was increased up to 91.67% after processing of samples. Conclusion: Sensitivity of smear microscopy can be enhanced by use of fluroscence microscopy and concentration method.展开更多
AIM:To explore an efficient,practical and objective quantitative method to evaluate the retinal neovascularization in mouse model of oxygen induced retinopathy (OIR).METHODS:Thirty C57BL/6J mice were explored in OIR m...AIM:To explore an efficient,practical and objective quantitative method to evaluate the retinal neovascularization in mouse model of oxygen induced retinopathy (OIR).METHODS:Thirty C57BL/6J mice were explored in OIR model procedure.Eyes were removed for different staining methods including:(1) HE staining;(2) immunohistochemistry with Griffonia Simplicifolia Lectin(GSL);(3) Immunofluore-scence with FITC labeled CD31 antibody;(4) Two-step immunofluorescence with purified-CD31 antibody;(5) FITC-Dextran perfusion combined with two-step purified-CD31immunofluorescence.Images of the retinal vasculature were analyzed by imaging software.RESULTS:GSL immunohistochemistry could clearly demonstrate the deep and superficial capillary beds.FITC labeled CD31 Immunofluorescence was blurring with high fluorescence background which was hard to distinguish retinal neovascularization in some area.Excellent detail of neovas-cularization and preexistent retinal vessels was provided in two-step Purified-CD31 immunofluorescence group.CONCLUSION:GSL immunohistochemistry can clearly demonstrate neovascularization tufts in deep and superficial capillary beds.Immunofluorescence of specific antigen CD31 on vascular endothelium can selectively label the neovascularization of mouse retina.When combined with computer analysis software,it is an effective and objective quantitative method to evaluate the retinal neovascularization in OIR mouse model.展开更多
We explored the discoloration of rattan cane using X-ray photoelectron spectroscopy(XPS) and Fourier transform infrared spectroscopy(FTIR). XPS analysis showed that after the cane was stained by Lasiodiplodia theobrom...We explored the discoloration of rattan cane using X-ray photoelectron spectroscopy(XPS) and Fourier transform infrared spectroscopy(FTIR). XPS analysis showed that after the cane was stained by Lasiodiplodia theobromae, carbon and oxygen elements and the ratio of oxygen to carbon decreased. Considering atomic binding,C_1 and C_4 contents increased, while C_2 and C_3 contents decreased, and the ratio of O_2 to O_1 decreased sharply. The relative contents of lignin, cellulose and polysaccharides increased and new substances with low O_2/O_1 ratio occurred. FTIR analysis showed that the absorption peaks of O–H at 3346 cm^(-1), aliphatic C–H at 2921, 2853 and1464 cm^(-1), and C=O at 1723 cm^(-1), were characteristic peaks of fungal melanin intensified, indicating that cane discoloration was primarily caused by fungal melanin. The absorption peaks characterizing cellulose and lignin like polysaccharides at 800 cm^(-1), C–H at 1374 cm^(-1), C–O at1058 and 1038 cm^(-1), phenolic hydroxyl at 1245 cm^(-1),aromatic ether bonds at 1270 cm^(-1), carbon skeleton at1608 cm^(-1) and benzene ring at 1500 cm^(-1) were enhanced since the fungus mainly consumed the extractives in cane cell lumens and the main composition content increased relatively. Regardless of the discoloration caused by natural fungi or inoculated fungi, the discoloring feature and composition changes were identical except that the fungusinoculated cane had more melanin.展开更多
AIM:To explore the value of Prussian blue staining in the diagnosis of ocular siderosis.METHODS:Between January 2012 and January 2013,the Prussian blue stain used in anterior lens capsule and vitreous liquid after cen...AIM:To explore the value of Prussian blue staining in the diagnosis of ocular siderosis.METHODS:Between January 2012 and January 2013,the Prussian blue stain used in anterior lens capsule and vitreous liquid after centrifugation from patients with definitive diagnosis and suspicious diagnosed of ocular siderosis. At the same time, give a negative control.RESULTS:Anterior lens capsule membrane and liquid of vitreous cavity from patients with definitive diagnosis and suspicious diagnosed of ocular siderosis revealed ferric ions that stained positively with Prussian blue. In the control group, there is no positive reaction.CONCLUSION:Prussian blue staining in the diagnosis of ocular siderosis has a very significant worth,suspected cases can be definitive diagnosed.展开更多
Objective To evaluate senile plaque formation and compare the sensitivity of three differentβ-amyloid(Aβ)labeling methods(antibody staining,Gallyas silver staining,and thioflavin-S staining)to detect Aβdeposition.M...Objective To evaluate senile plaque formation and compare the sensitivity of three differentβ-amyloid(Aβ)labeling methods(antibody staining,Gallyas silver staining,and thioflavin-S staining)to detect Aβdeposition.Methods APPswe/PSEN1dE9 transgenic mice(APP/PS1)of different ages were used to examine spatiotemporal changes in Aβplaque deposition.Antibody staining,Gallyas silver staining,and thioflavin-S staining were used to detect Aβplaque deposition in the same brain region of adjacent slices from model mice,and the results were compared.Results With aging,Aβplaques first appeared in the cortex and then the deposition increased throughout the whole brain.Significantly greater plaque deposition was detected by 6E10 antibody than that analyzed with Gallyas silver staining or thioflavin-S staining(P<0.05).Plaque deposition did not show significant difference between the APP/PS1 mice brains assayed with Gallyas silver staining and ones with thioflavin-S staining(P=0.0033).Conclusions The APP/PS1 mouse model of Alzheimer’s disease could mimick the progress of Aβplaques occurred in patients with Alzheimer’s disease.Antibody detection of Aβdeposition may be more sensitive than chemical staining methods.展开更多
Polygala paniculata L.is a medicinal plant that grows in the Brazilian Atlantic coast,known as‘barba-de-São-João’,‘barba-de-bode’,‘vassourinha branca’,and‘mimosa’.In this study,pollen viability was e...Polygala paniculata L.is a medicinal plant that grows in the Brazilian Atlantic coast,known as‘barba-de-São-João’,‘barba-de-bode’,‘vassourinha branca’,and‘mimosa’.In this study,pollen viability was estimated by three different staining methods:2%acetic orcein,2%acetic carmine,and Alexander’s stain.The young inflorescences of twenty accessions were collected and fixed in a solution of ethanol:acetic acid(3:1)for 24 hours,then stored in ethanol 70%under refrigeration.Six slides per plant,two for each stain,were prepared by squashing,and 300 pollen grains per slide were analyzed.Pollen viability was high(>70%)for most accessions of P.paniculata using the Alexander’s stain,which proved the most adequate method to estimate pollen viability.展开更多
In an age where digitization is widespread in clinical and preclinical workflows,pathology is still predominantly practiced by microscopic evaluation of stained tissue specimens affixed on glass slides.Over the last d...In an age where digitization is widespread in clinical and preclinical workflows,pathology is still predominantly practiced by microscopic evaluation of stained tissue specimens affixed on glass slides.Over the last decade,new high throughput digital scanning microscopes have ushered in the era of digital pathology that,along with recent advances in machine vision,have opened up new possibilities for Computer-Aided-Diagnoses.Despite these advances,the high infrastructural costs related to digital pathology and the perception that the digitization process is an additional and nondirectly reimbursable step have challenged its widespread adoption.Here,we discuss how emerging virtual staining technologies and machine learning can help to disrupt the standard histopathology workflow and create new avenues for the diagnostic paradigm that will benefit patients and healthcare systems alike via digital pathology.展开更多
文摘Stains are applied to impart contrast to the tissue and identify particular features of interest. However, the use of synthetic dyes as staining reagents has been associated with significant human health challenges and pollution of the ecosystem. These developments have necessitated a shift towards using natural dyes that are eco-friendlier and readily available. We investigated the staining reaction patterns of teak tree leaves (Tectona grandis) dye extracts and explored their suitability as a cytoplasmic stain in micromorphological assessments. Dye extracts were prepared using acetone, methanol, and ethanol as solvents from air-dried (under shade) teak tree young leaves. The dye extracts were applied as a counterstain and evaluated against eosin in formalin-fixed paraffin-embedded (FFPE) bovine tissue sections at varying concentrations and different staining times. Teak tree leaves (Tectona grandis) dye extracts produced relatively varying staining intensities of reddish-brown cytoplasmic coloration when used on bovine tissue at different concentrations and staining times comparable to eosin and with blue-purple hematoxylin nuclear stain. The present study showed that Tectona grandis leaf dye extracts provide an excellent cytoplasmic staining pattern and can be used as an alternative counterstain in routine H&E staining techniques.
基金National Natural Science Foundation of China,Grant/Award Number:81730031 to lili and 82001395 to yingwei wangthe Foundation of Shanghai Municipal Key Clinical Specialty,Grant/Award Number:shslczdzk06901 to yingwei wang。
文摘Background:TTC(2,3,5-triphenyltetrazolium chloride)staining is the most commonly used method in identifying and assessing cerebral infarct volumes in the transient middle cerebral artery occlusion model.Given that microglia exhibit different morphologies in different regions after ischemic stroke,we demonstrate the superiority and necessity of using TTC-stained brain tissue to analyze the expression of various proteins or genes in different regions based on microglia character.Methods:We compared brain tissue(left for 10 min on ice)from the improved TTC staining method with penumbra from the traditional sampling method.We identified the feasibility and necessity of the improved staining method using real time(RT)-PCR,Western blot,and immunofluorescence analysis.Results:There was no protein and RNA degradation in the TTC-stained brain tissue group.However,the TREM2 specifically expressed on the microglia showed a significant difference between two groups in the penumbra region.Conclusions:TTC-stained brain tissue can be used for molecular biology experiments without any restrictions.In addition,TTC-stained brain tissue shows greater superiority due to its precise positioning.
文摘Introduction: This study aimed to perform routine seminal fluid analysis, sperm DNA fragmentation, and sperm function tests at the chromatin maturation level and evaluate pregnancy in the patients passing intrauterine insemination before starting Intrauterine Insemination (IUI) method. Materials and Methods: In this prospective study, 111 couples who underwent Intrauterine Insemination (IUI) in unexplained infertility patients were admitted to Al-Farah IVF and assisted reproductive center in Baghdad, Iraq between November 2020 and February 2021 were evaluated. Semen fluid analysis was performed based on (WHO 4th) guiding rules. In addition, Sperm Chromatin Dispersion (halo test) and sperm maturation were performed with Aniline Blue Stain (ABS). Results: Sperm Chromatin Dispersion (SCD) groups were compared in terms of pregnancy outcome;the positive pregnancy rate was found to be above in the normal SCD groups (p = 0.0005). In addition, Aniline Blue Stain (ABS) groups were compared in the terms of pregnancy outcome;the positive pregnancy rate was found to be higher in the normal ABS group (p = 0.017). Conclusion: Our study showed that the use of DNA fragmentation (SCD) and sperm maturation tests (ABS) together with routine semen analysis in intrauterine insemination cases will make a significant contribution to the prediction of Intrauterine Insemination (IUI) increased results. So, these results indicate a defect in the effect of DNA fragmentation on the outcome of intrauterine insemination.
基金the National Natural Science Foundation of China(grant no.81971847)Interdisciplinary Program of Shanghai Jiao Tong University(grant no.YG2019QNB10)+1 种基金Shanghai Municipal Key Clinical Specialty(grant no.shslczdzk00901)Shanghai Municipal Commission of Health and Family Planning(grant no.202240150)。
文摘Port-wine stain(PWS)is a congenital capillary malformation that occurs in 0.3%–0.5%of newborns.The pulsed dye laser is the current gold standard treatment for PWS;however,its efficacy is poor.Photosensitizer photodynamic therapy(PDT)is considered a promising treatment for PWS.Here we provide a comprehensive overview of PDT.
基金supported by an unrestricted grant from Research to Prevent Blindness and NIH grants EY14801,EY031292.
文摘In the early days of deciphering the injured neuronal tissues led to the realization that contrast is necessary to discern the parts of the recovering tissues from the damaged ones.Early attempts relied on available(and often naturally occurring)staining substances.Incidentally,the active ingredients of most of them were small molecules.With the advent of time,the knowledge of chemistry helped identify compounds and conditions for staining.The staining reagents were even found to enhance the visibility of the organelles.Silver impregnation identification of Golgi bodies was discovered in owl optic nerve.Staining reagents since the late 1800s were widely used across all disciplines and for nerve tissue and became a key contributor to advancement in nerve-related research.The use of these reagents provided insight into the organization of the neuronal tissues and helped distinguish nerve degeneration from regeneration.The neuronal staining reagents have played a fundamental role in the clinical research facilitating the identification of biological mechanisms underlying eye and neuropsychiatric diseases.We found a lack of systematic description of all staining reagents,whether they had been used historically or currently used.There is a lack of readily available information for optimal staining of different neuronal tissues for a given purpose.We present here a grouping of the reagents based on their target location:(I)the central nervous system(CNS),(II)the peripheral nervous system(PNS),or(III)both.The biochemical reactions of most of the staining reagents is based on acidic or basic pH and specific reaction partners such as organelle or biomolecules that exists within the given tissue type.We present here a summary of the chemical composition,optimal staining condition,use for given neuronal tissue and,where possible,historic usage.Several biomolecules such as lipids and metabolites lack specific antibodies.Despite being non-specific the reagents enhance contrast and provide corroboration about the microenvironment.In future,these reagents in combination with emerging techniques such as imaging mass spectrometry and kinetic histochemistry will validate or expand our understanding of localization of molecules within tissues or cells that are important for ophthalmology and vision science.
文摘Introduction: Reports indicate that fluorescent staining of smears increases sensitivity of direct microscopy;so ZN staining is being replaced with fluorescent microscopy in RNTCP in India. Chemical processing and sputum concentration may also improve sensitivity of microscopy. Objective: To compare the sensitivity and specificity of microscopy for AFB using ZN and fluorescent stains in direct and concentrated specimen with culture as gold standard. Methods: Morning sputum specimen of patients, suspected of having pulmonary tuberculosis, over a period of 6 months was subjected to direct microscopy using fluorescent stain;the same slide was over-stained with ZN stain. Same sputum sample was concentrated by Petroff’s method and subjected to fluorescent microscopy followed by ZN microscopy and finally to culture for AFB. Results: Sensitivity of fluorescent stained concentrated sputum samples was maximum and of ZN stained unprocessed sputum samples was minimum. Specificity of three of the methods was equal at 0.96 but of ZN stained concentrated sputum smears was 0.97. Sensitivity of total fluorescent stains was 0.85 (Specificity 0.96) and sensitivity of total ZN stained smears was 0.80 (Specificity 0.96). Discussion: We used same smear for fluorescent and ZN stains, so smear related variability is decreased. Blinding for microscopy was practically complete. Conclusion: The sensitivity of sputum microscopy for AFB can be increased by concentrating the sputum and using fluorescent microscopy. The specificity remains high in all the methods.
文摘In order to find a simple and conve-nient method of diagnosis and to probe intothe value of auricular diagnosis in malignanttumors so as to substantiate and
文摘Negative staining is an effective method that can be used for electron microscopic study to observe fine structural morphology without destruction of bacterial structure. Although uranium acetate is used worldwide as a general dyeing solution, it is extremely difficult to use it by a new purchase at a research institution because it falls under the nuclear regulation substance in Japan. Therefore, we examined alternative reagents for negative staining that could replace uranium acetate through bacterial observation with an electron microscope. Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, and Streptococcus pyogenes were examined by four stain reagents (phosphotungstic acid (PTA), EMstainer, TI blue, and uranium acetate). Pre cultured bacteria were stained with each stain reagents on a copper grid, washed with PBS, and observed with a transmission electron microscope. In the comparison between bacterial structures, the cell wall structure and bacterial flagella could be observed well in the order of PTA, EMstainer, and uranium acetate. With TI blue staining, flagella could be observed very poorly. In comparison between bacteria, gram negative bacteria such as Escherichia coli and Pseudomonas aeruginosa, could be observed well as compared with gram positive cocci such as Staphylococcus aureus and Streptococcus pyogenes. The uranium acetate looked very coarse in background particles. Since crystals tend to precipitate, TI blue also required filtering, and electron beams were absorbed by the agglomerated crystals, and the frequency of electronic burning occurred high frequency. In this study, there was clear difference in the observation conditions depending on the type of bacteria and the kind of the staining reagents. Especially, it was confirmed that good negative staining features of Pseudomonas aeruginosa by electron microscope were obtained by PTA and EMstainer staining. These alternative reagents are considered to be a candidate for a negative staining.
基金Liaoning Natural Science Foundation:Research on functional dyes for intraocular biofilms(No.20180550004)Shenyang Science and Technology Plan Project:Preparation of Chitosan Bioactive Dressing and Study on Reconstruction of Bulbular Conjunctiva(No.19-112-4-069)Liaoning Provincial Natural Science Foundation:A study on the role of a degradable bioactive dressing in the repair of bulbar conjunctiva(No.2019-MS-257)
文摘Objective: To explore the effect of gardenia blue pigment on the anterior capsule staining and its safety performance. Methods: In this study, New Zealand white rabbits were used as the research object to test the effect of gardenia blue pigment on the anterior capsule staining and anterior segment toxicity. In this study, gardenia blue stains of different concentrations (5%, 4%, 3%, 2%, 1%, 0.5%) were prepared to stain the anterior capsule of rabbit crystals, and the effect of the stain was observed to determine the optimal concentration of the stain. Twenty-seven healthy New Zealand white rabbits without eye disease were randomly divided into three groups: gardenia blue staining group;bio-blue positive drug group;physiological saline group. The intraocular pressure, anterior chamber inflammation, corneal endothelial injury, and anterior chamber angle pathological changes were measured in 1D, 7D, and 14D. Results: The results of this study showed that the effect of gardenia blue pigment on the anterior capsule of the lens was the best when the concentration of blue pigment was 2%. There was no change in intraocular pressure in the anterior chamber for a short period of time. There is no damage to the corneal endothelium. Conclusion: The results of this study show that the effect of gardenia blue pigment on the anterior capsule is best when the concentration of blue pigment is 2%, and the stain has no obvious toxic and side effects on the anterior segment of the eye.
文摘Stains and staining methods significantly assist in diagnoses in medical research and health care.Certain color of a dye can identify the location of a tumor within a specimen.Applying histochemical-staining enabled morphological identification of fibrin in the lymphoid tissue during cancer progression.Staining methods in combination with the LSFM(light-sheet fluorescence microscopy)allowed tracing the drug penetration,development and spread of tumors.The Curcumin dye is in use for labelling and imaging of Aβplaques in post-mortem brain tissue.Immunofluorescent staining methods are employed in detection of some important proteins in early diagnostic changes relevant to heart damage.The methods are developed in medical research to include stem cells and tissue engineering,cell culturesproperties and capabilities,connective tissues and extracellular matrix,nervous system,musculoskeletal system;respiratory system,liver and gastrointestinal tract,and male and female reproductive systems.
文摘One of the significant features of The Human Stain is that it lays bare the reality of American society and censures the Anglo-Saxon or white discourse.What are equally important are the multiple narrative strategies that Philip R oth employs in the novel.T his paper focuses mainly on the elaboration of such narrative strategies as mimesis and diegesis,complicated narrative time,shifting focalizations,and narration or non-narrative comments,so as to point out that these narrative strategies add colors to its( postmodern) artistic features,becoming an integral component of this novel.
文摘Background: Tuberculosis is a highly infectious disease and India has the highest burden with it. Diagnosis of tuberculosis in many countries is still dependent on microscopy. Although its sensitivity is low in comparison to culture and molecular methods, its sensitivity can still be improved by using fluorescence staining method and processing of samples by homogenization and concentration method. Material and methods: Samples were collected from all newly registered suspected cases of tuberculosis in tertiary care hospital from outward and indoor department during a period of one year. Smears were prepared for Ziehl Neelsen stain and fluorescence stain both before and after homogenization and concentration procedure by 4% NAOH-2.9% sodium citrate method and results of them were interpreted according to RNTCP criteria for grading of sputum samples. All the samples were cultured in liquid culture MGIT system (Mycobacterial Growth Indicator Tube) and results of microscopy were compared with liquid culture taken as gold standard. Data were analyzed by using SPSS software version 16. Result: 350 samples were collected during study period. Out of 350 samples, 48 samples were positive for M. tuberculosis by MGIT system. In comparison with MGIT system, sensitivity of Z N stain for detection of acid fast bacilli was 77% before decontamination procedure, which was increased up to 85.42% after decontamination and concentration process. Sensitivity of fluroscence stain was 85.42% before processing, which was increased up to 91.67% after processing of samples. Conclusion: Sensitivity of smear microscopy can be enhanced by use of fluroscence microscopy and concentration method.
基金Supported by National Natural Science Foundation of China(No.30973899)
文摘AIM:To explore an efficient,practical and objective quantitative method to evaluate the retinal neovascularization in mouse model of oxygen induced retinopathy (OIR).METHODS:Thirty C57BL/6J mice were explored in OIR model procedure.Eyes were removed for different staining methods including:(1) HE staining;(2) immunohistochemistry with Griffonia Simplicifolia Lectin(GSL);(3) Immunofluore-scence with FITC labeled CD31 antibody;(4) Two-step immunofluorescence with purified-CD31 antibody;(5) FITC-Dextran perfusion combined with two-step purified-CD31immunofluorescence.Images of the retinal vasculature were analyzed by imaging software.RESULTS:GSL immunohistochemistry could clearly demonstrate the deep and superficial capillary beds.FITC labeled CD31 Immunofluorescence was blurring with high fluorescence background which was hard to distinguish retinal neovascularization in some area.Excellent detail of neovas-cularization and preexistent retinal vessels was provided in two-step Purified-CD31 immunofluorescence group.CONCLUSION:GSL immunohistochemistry can clearly demonstrate neovascularization tufts in deep and superficial capillary beds.Immunofluorescence of specific antigen CD31 on vascular endothelium can selectively label the neovascularization of mouse retina.When combined with computer analysis software,it is an effective and objective quantitative method to evaluate the retinal neovascularization in OIR mouse model.
基金supported by the National Key Project of S&T Supporting Programs Funded by MOST of China during the 12th Five-year Plan(No.2012BAD23B0104)
文摘We explored the discoloration of rattan cane using X-ray photoelectron spectroscopy(XPS) and Fourier transform infrared spectroscopy(FTIR). XPS analysis showed that after the cane was stained by Lasiodiplodia theobromae, carbon and oxygen elements and the ratio of oxygen to carbon decreased. Considering atomic binding,C_1 and C_4 contents increased, while C_2 and C_3 contents decreased, and the ratio of O_2 to O_1 decreased sharply. The relative contents of lignin, cellulose and polysaccharides increased and new substances with low O_2/O_1 ratio occurred. FTIR analysis showed that the absorption peaks of O–H at 3346 cm^(-1), aliphatic C–H at 2921, 2853 and1464 cm^(-1), and C=O at 1723 cm^(-1), were characteristic peaks of fungal melanin intensified, indicating that cane discoloration was primarily caused by fungal melanin. The absorption peaks characterizing cellulose and lignin like polysaccharides at 800 cm^(-1), C–H at 1374 cm^(-1), C–O at1058 and 1038 cm^(-1), phenolic hydroxyl at 1245 cm^(-1),aromatic ether bonds at 1270 cm^(-1), carbon skeleton at1608 cm^(-1) and benzene ring at 1500 cm^(-1) were enhanced since the fungus mainly consumed the extractives in cane cell lumens and the main composition content increased relatively. Regardless of the discoloration caused by natural fungi or inoculated fungi, the discoloring feature and composition changes were identical except that the fungusinoculated cane had more melanin.
基金Supported by Education Department Funding of Sichuan Province,China(No.2005B020)
文摘AIM:To explore the value of Prussian blue staining in the diagnosis of ocular siderosis.METHODS:Between January 2012 and January 2013,the Prussian blue stain used in anterior lens capsule and vitreous liquid after centrifugation from patients with definitive diagnosis and suspicious diagnosed of ocular siderosis. At the same time, give a negative control.RESULTS:Anterior lens capsule membrane and liquid of vitreous cavity from patients with definitive diagnosis and suspicious diagnosed of ocular siderosis revealed ferric ions that stained positively with Prussian blue. In the control group, there is no positive reaction.CONCLUSION:Prussian blue staining in the diagnosis of ocular siderosis has a very significant worth,suspected cases can be definitive diagnosed.
基金Supported by the 2016 Major Collaborative Innovation Program of the Chinese Academy of Medical Sciences(2016-I2M-1004)
文摘Objective To evaluate senile plaque formation and compare the sensitivity of three differentβ-amyloid(Aβ)labeling methods(antibody staining,Gallyas silver staining,and thioflavin-S staining)to detect Aβdeposition.Methods APPswe/PSEN1dE9 transgenic mice(APP/PS1)of different ages were used to examine spatiotemporal changes in Aβplaque deposition.Antibody staining,Gallyas silver staining,and thioflavin-S staining were used to detect Aβplaque deposition in the same brain region of adjacent slices from model mice,and the results were compared.Results With aging,Aβplaques first appeared in the cortex and then the deposition increased throughout the whole brain.Significantly greater plaque deposition was detected by 6E10 antibody than that analyzed with Gallyas silver staining or thioflavin-S staining(P<0.05).Plaque deposition did not show significant difference between the APP/PS1 mice brains assayed with Gallyas silver staining and ones with thioflavin-S staining(P=0.0033).Conclusions The APP/PS1 mouse model of Alzheimer’s disease could mimick the progress of Aβplaques occurred in patients with Alzheimer’s disease.Antibody detection of Aβdeposition may be more sensitive than chemical staining methods.
文摘Polygala paniculata L.is a medicinal plant that grows in the Brazilian Atlantic coast,known as‘barba-de-São-João’,‘barba-de-bode’,‘vassourinha branca’,and‘mimosa’.In this study,pollen viability was estimated by three different staining methods:2%acetic orcein,2%acetic carmine,and Alexander’s stain.The young inflorescences of twenty accessions were collected and fixed in a solution of ethanol:acetic acid(3:1)for 24 hours,then stored in ethanol 70%under refrigeration.Six slides per plant,two for each stain,were prepared by squashing,and 300 pollen grains per slide were analyzed.Pollen viability was high(>70%)for most accessions of P.paniculata using the Alexander’s stain,which proved the most adequate method to estimate pollen viability.
基金This study was financially supported by the NSF Biophotonics Program (USA).
文摘In an age where digitization is widespread in clinical and preclinical workflows,pathology is still predominantly practiced by microscopic evaluation of stained tissue specimens affixed on glass slides.Over the last decade,new high throughput digital scanning microscopes have ushered in the era of digital pathology that,along with recent advances in machine vision,have opened up new possibilities for Computer-Aided-Diagnoses.Despite these advances,the high infrastructural costs related to digital pathology and the perception that the digitization process is an additional and nondirectly reimbursable step have challenged its widespread adoption.Here,we discuss how emerging virtual staining technologies and machine learning can help to disrupt the standard histopathology workflow and create new avenues for the diagnostic paradigm that will benefit patients and healthcare systems alike via digital pathology.
