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 O...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) Immunofluorescence 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 neovascularization 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
In this study, mitotic metaphase chromosomes in mouse were identified by a new chromosome fluorescence banding technique combining DAPI staining with image analysis. Clear 4', 6-diamidino-2-phenylindole (DAPI) mult...In this study, mitotic metaphase chromosomes in mouse were identified by a new chromosome fluorescence banding technique combining DAPI staining with image analysis. Clear 4', 6-diamidino-2-phenylindole (DAPI) multiple bands like (J-hands could be produced in mouse. The Meta- Morph software was then used to generate linescans of pixel intensity for the banded chromosomes from short arm to long arm. These linescans were sufficient not only to identify each individual chromosome but also analyze the physical sites of bands in chromosome. Based on the results, the clear and accurate karyotype of mouse metaphase chromosomes was established. The technique is therefore considered to he a new method for cytological studies of mouse.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
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
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
Objective:The recent advent of flow cytometry(FCM),coupled with fluorescent dyes,has been successfully applied to assess mitochondrial function.The aim of this study was to investigate the feasibility and clinical ...Objective:The recent advent of flow cytometry(FCM),coupled with fluorescent dyes,has been successfully applied to assess mitochondrial function.The aim of this study was to investigate the feasibility and clinical significance of detecting sperm mitochondrial function and to evaluate sperm mitochondrial function by using Rhodamine 123/propidium(Rh123/PI)dual fluorescent staining and FCM in asthenospermia and oligoasthenozoospermia.Methods:Twenty-five fertile men(with normal sperm parameters)and 230 infertile patients were examined.Fifty-five patients of the above 230 patients were selected for idiopathic infertility samples and were divided into two groups:asthenospermia(n=30)and oligoasthenozoospermia(n=25).Rh123/PI dual fluorescent staining and FCM were carried out to examine sperm mitochondrial function.Results:Significant differences were found between the normal and abnormal semen samples(P0.05)when Rh123+/PI-,Rh123-/PI+and Rh123-/PI-sperm were examined by FCM,but there was no significant difference between the asthenospermia(P=0.469) and oligoasthenozoospermia group(P=0.950)when Rh123+/PI-and Rh123-/PI+sperm were then examined;however,a significant difference was found between the 2 groups(P=0.003)when Rh123-/PI-sperm were examined.There was no correlation between Rh123-/PI-sperm and semen parameters in the normal group,but there was a significant negative correlation between the sperm concentration and Rh123-/PI-sperm in asthenospermia and oligoasthenozoospermia patients(r=-0.509,-0.660;P=0.018,0.038).Conclusion:Rh123/PI dual fluorescent staining and FCM can provide reliable information to assess the quality of sperm and reveal differences in mitochondrial membrane potential in asthenospermia and oligoasthenozoospermia.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
Microbial activity in the water injection system in oil and gas industry leads to an array of challenges, including biofouling, injectivity loss, reservoir plugging, and microbiologically influenced corrosion (MIC). A...Microbial activity in the water injection system in oil and gas industry leads to an array of challenges, including biofouling, injectivity loss, reservoir plugging, and microbiologically influenced corrosion (MIC). An effective mitigation strategy requires online and real-time monitoring of microbial activity and growth in the system so that the operators can apply and adjust counter-measures quickly and properly. The previous study [1] identified DNA staining technology-with PicoGreen and SYBR Green dyes—as a very promising method for automated, online determination of microbial cell abundance in the vast Saudi Aramco injection seawater systems. This study evaluated DNA staining technology on detection limit, automation potential, and temperature stability for the construction of automated sensor prototype. DNA staining with SYBR Green dye was determined to be better suited for online and real-time monitoring of microbial activity in the Saudi Aramco seawater systems. SYBR Green staining does not require sample pre-treatment, and the fluorescence signal intensity is more stable at elevated temperatures up to 30℃. The lower detection limit of 2 × 10<sup>3</sup>/ml was achieved under the optimized conditions, which is sufficient to detect microbial numbers in Saudi Aramco injection seawater. Finally, the requirements for design and construction of SYBR-based automated sensor prototype were determined.展开更多
Using potted seedlings of Chlorophytum comosum var. variegatum as the experimental materials, the effect of 2.0 mmol/L methyl orange ( Treatment T1 ), 1.0 mmol/L methyl violet ( Treatment T2 ) and 1.0 mmol/L neutr...Using potted seedlings of Chlorophytum comosum var. variegatum as the experimental materials, the effect of 2.0 mmol/L methyl orange ( Treatment T1 ), 1.0 mmol/L methyl violet ( Treatment T2 ) and 1.0 mmol/L neutral red ( Treatment T3 ) on the biomass, root-shoot ratio, leaf color indices, plant carbon and nitrogen nutrition were studied. The results showed that the biomass of Treatment T3 was significantly greater than that of treatments T1 and CK. The root-shoot ratio decreased significantly in treatments T1, T2 and T3 , and the decrease in T3 was most obvious. In all the three treatments with coloring agent, a ^* , b ^* and L ^* values were increased gradually, C value were decreased, H0 and CIRG were increased, and the leaves were pink. In addition, the contents of chlorophyll a, chlorophyll b, chlorophyll a + b and carotenoid were significantly decreased. The contents of soluble sugar and starch were also decreased, and the decrease in Treatment T2 was most significant. The contents of soluble protein and total nitrogen were increased, and the increase was most dramatic in Treatment T3. The carbon to nitrogen ratio was decreased. The results proved that staining can improve the ornamental value of indoor plants, despite its effects on plant carbon and nitrogen nutrition of C. comosum vat. variegatum, dyeing.展开更多
A detailed morphological study of neurons in healthy and pathological conditions requires reasonably a number of special techniques, which may visualize the majority of neu- rons in a thick three-dimensional arrangeme...A detailed morphological study of neurons in healthy and pathological conditions requires reasonably a number of special techniques, which may visualize the majority of neu- rons in a thick three-dimensional arrangement. A detailed visualization of neurons must include the cell body, most of the dendritic arbor, the dendritic spines, the axon, the axonal collaterals and the synapses. An ideal morphological technique for the study of degeneration and regeneration processes of the central nervous system must also visualize clearly the long and short neuronal circuits, as well as the dendritic and axonal bands and tracks.展开更多
The solution chemical and optical characteristics of formation of amine-terminated polyamidoamine dendrimer G2.0(NH2-PAMAM G2.0)-Au nanocomposites in the aqueous solution of NH2-PAMAM G2.0 at various mole ratios of...The solution chemical and optical characteristics of formation of amine-terminated polyamidoamine dendrimer G2.0(NH2-PAMAM G2.0)-Au nanocomposites in the aqueous solution of NH2-PAMAM G2.0 at various mole ratios of Au(Ⅲ) to NH2-PAMAM G2.0 were studied by both UV-visible spectrometry and fluorospectrometry. The NH2-PAMAM G2.0-Au nanocomposites, with a type of structure in which one Au nanoparticle is surrounded by several NH2-PAMAM G2.0 dendrimers, emit strong bluish violet fluorescence, and are uniform, water soluble and biocompatible as well as very stable in frozen conditions. The size of gold nanoparticles in the nanocomposites is about 2.5 nm and decreases with the increase of NH2-PAMAM G2.0 concentration. The NH2-PAMAM G2.0 plays an important role in acting as host or micro-reactor for Au(Ⅲ) before Au(Ⅲ) reduction and acting as dispersant and stabilizer for gold nanoparticles after Au(Ⅲ) reduction. Preliminary experiments of cells staining to human embryonic lung fibroblast cell lines show that the NH2-PAMAM G2.0-Au nanocomposites can be used as optical imaging markers for bioanalyses and medical diagnoses.展开更多
AIM: To evaluate clinicopathologic parameters and the clinical significance related lymphovascular invasion (LVI) by immunohistochemical staining (IHCS) in endoscopic submucosal dissection (ESD). METHODS: Between May ...AIM: To evaluate clinicopathologic parameters and the clinical significance related lymphovascular invasion (LVI) by immunohistochemical staining (IHCS) in endoscopic submucosal dissection (ESD). METHODS: Between May 2005 and May 2010, a total of 348 lesions from 321 patients (mean age 63 ± 10 years, men 74.6%) with early gastric cancer (EGC) who met indication criteria after ESD were analyzed retrospectively. The 348 lesions were divided into the absolute (n = 100, differentiated mucosal cancer without ulcer ≤ 20 mm) and expanded (n = 248) indica-tion groups after ESD. The 248 lesions were divided into four subgroups according to the expanded ESD indication. The presence of LVI was determined by factor Ⅷ-related antigen and D2-40 assessment. We compared LVI IHCS-negative group with LVI IHCSpositive in each group. RESULTS: LVI by hematoxylin-eosin staining (HES) and IHCS were all negative in the absolute group, while was observed in only the expanded groups. The positive rate of LVI by IHCS was higher than that of LVI by HES (n = 1, 0.4% vs n = 11, 4.4%, P = 0.044). LVI IHCS-positivity was observed when the cancer invaded to the mucosa 3 (M3) or submucosa 1 (SM1) levels, with a predominance of 63.6% in the subgroup that included only SM1 cancer (P < 0.01). In a univariate analysis, M3 or SM1 invasion by the tumor was significantly associated with a higher rate of LVI by IHCS, but no factor was significant in a multivariate analysis. There were no cases of tumor recurrence or metastasis during the median 26 mo follow-up. CONCLUSION: EGCs of the absolute group are immunohistochemically stable. The presence of LVI may be carefully examined by IHCS in an ESD expanded indication group with an invasion depth of M3 or greater.展开更多
Fast Plant(Brassica rapa,Cruciferae)leaf tissuefixed in glutaraldehyde-acrolein and post-fixed in os-mium,was examined for response to several easily-prepared heavy metal stains.Lead and uranium,separately and in comb...Fast Plant(Brassica rapa,Cruciferae)leaf tissuefixed in glutaraldehyde-acrolein and post-fixed in os-mium,was examined for response to several easily-prepared heavy metal stains.Lead and uranium,separately and in combination,gave typical resultsacross the spectrum of cell organellets.As a single stainfollowing osmium,bismuth produced images seeminglyequivalent to lead and uranium.Phosphotungstic acidproduced very good membrane delineation but produceda washed-out background image similar to that from leadstaining.Carbohydrate compounds were especiallyresponsive to ruthenium;the cytoplasm and the matrixof all organelles were also stained very well.Theprocedures were no more demanding than traditionalstaining methods and may be easily used in research andteaching.Fast Plant materials are a reliable,quick andeasy source of living material.展开更多
EST isozymes are one of the frequently used biochemical markers in genetic analysis of fungi. and the staining is an important process in electrophoresis analysis of studying Est.The effects were compared ainong diff...EST isozymes are one of the frequently used biochemical markers in genetic analysis of fungi. and the staining is an important process in electrophoresis analysis of studying Est.The effects were compared ainong different stain recipes for Est of 3 kinds of fungi-Lentinus edodes. Pleurotus sapidus. Phellinus igriarius and 2 kinds of plants-Populus sp and Brassica chinensis. Of the four kinds of Est staining recipes tested.the recipe α-acetic acid-naphther showed the best effect.and followed by β-aceticacid-naphther, semicontent α-aceticacid-naphther and α+β-aceticacidnathpher.展开更多
AIM: To investigate whether performing immuno-histochemical CD3 staining, in order to improve the detection of intra-epithelial lymphocytosis, has an additional value in the histological diagnosis of celiac disease.ME...AIM: To investigate whether performing immuno-histochemical CD3 staining, in order to improve the detection of intra-epithelial lymphocytosis, has an additional value in the histological diagnosis of celiac disease.METHODS: Biopsies obtained from 159 children were stained by hematoxylin and eosin(HE) and evaluated using the Marsh classification. CD3 staining was subsequently evaluated separately and independently.RESULTS: Differences in evaluation between the routine HE sections and CD3 staining were present in 20(12.6%) cases. In 10(6.3%) patients the diagnosis of celiac disease(Marsh Ⅱ and Ⅲ) changed on examination of CD3 staining: in 9 cases, celiac disease had initially been missed on the HE sections, while 1 patient had been over-diagnosed on the routine sections. In all patients, the final diagnosis based on CD3 staining, was concordant with serological results, which was not found previously. In the other 10(12.3%) patients, the detection of sole intra-epithelial lymphocytosis(Marsh Ⅰ) improved. Nine patients were found to have Marsh Ⅰ on CD3 sections, which had been missed on routine sections. Interestingly, the only patient with negative serology had Giardiasis. Finally, in 1 patient with negative serology, in whom Marsh Ⅰ was suspected on HE sections, this diagnosis was withdrawn after evaluation of the CD3 sections.CONCLUSION: Staining for CD3 has an additional value in the histological detection of celiac disease lesions, and CD3 staining should be performed when there is a discrepancy between serology and the diagnosis made on HE sections.展开更多
BACKGROUND: There are many methods for myelin staining, mordant, or oil-soluble dye or the special reaction of osmic acid with lipoid is used according to different principles. The commonly used methods are classic We...BACKGROUND: There are many methods for myelin staining, mordant, or oil-soluble dye or the special reaction of osmic acid with lipoid is used according to different principles. The commonly used methods are classic Well staining, classic lithium carbonate-haematine staining, fast green staining, silver staining, etc. Luxol Fast Blue can brightly stain myelin sheath, and has certain specificity. The background can be very clean if there is proper differentiation, whereas Luxol Fast Blue is cheap and convenient to operate, thus it is an ideal staining reagent for routine myelin sheath. OBJECTIVE: To show the corticospinal tract of normal adult rats with Luxol Fast Blue staining method. DESIGN: A repetitive measurement design. SETTINGS: Institute of Nuerobiology, Nantong University; Department of Rehabilitation Medicine, Affiliated Hospital of Nantong University. MATERIALS: Six healthy adult male SD rats of clean degree, weighing averagely 300 g, were provided by the experimental animal center of Nantong University. 1 g/L Luxol Fast Blue solution was provided by Sigma Company; Leica CM1900 cryostat microtome by Leica Company; Leica DMR microscope by Leica Company. METHODS: The experiment was carried out in the Staff Room of Human Anatomy, Nantong University in May 2005. The rats were given intraperitoneal injection of combined anesthetic (2 mL/kg), then the chest was open for perfusing saline and phosphate buffer containing formamint via heart. Brain and spinal cord were removed after 1 hour then fixed, then changed to phosphate buffer (pH 7.4) containing 300 g/L saccharu at 4 ℃, and stayed overnight, tissue blocks at pyramid, decussation of pyramid and cervical, thoracic, lumbar and sacral segments of spinal cord were removed to prepare continuous horizontal frozen sections (30 μm) after sedimentation, the sections were dried at room temperature. The corticospinal tract of normal adult rats were shown with Luxol Fast Blue staining method, and observed under Leica DMR microscope. MAIN OUTCOME MEASURES: Positive fibers in Luxol Fast Blue staining. RESULTS: After the Luxol Fast Blue staining, the labeled myelinated nerve fibers were bright blue. They located in the pyramid, decussation of pyramid and the ventral part of posterior funiculus in cervical, thoracic, lumbar and sacral segments of spinal cord. CONCLUSION: Luxol Fast Blue staining method may manifest the distribution of corticospinal tract with clear distinct in adult rats.展开更多
Fetal liver tissues obtained from 28 human fetuses with gestation age from 3 to 6 months and fetal bone marrow from 35 human fetuses from 3 to 7 months were observed by immunochemical staining with anti-platelet GPⅡ ...Fetal liver tissues obtained from 28 human fetuses with gestation age from 3 to 6 months and fetal bone marrow from 35 human fetuses from 3 to 7 months were observed by immunochemical staining with anti-platelet GPⅡ b / Ⅲa monoclonal antibody and ABC technique. In the fetal liver, megakaryocytes were wholly located among growing fetal liver cells and near foci of hemopoiesis. Some megakaryocytes in the fetal liver were small7890- lymphoid-like megakaryocytes. The size of megakaryocytes both in the fetal liver (14.79 ± 4.52μm) and in the fetal bone marrow (16.08±7.39 μm) was small, which did not vary significantly over the gestation age ranging from 3 to 6 or 7 months. However, the maturation stage of megakaryocytes in the fetal liver shifted to more mature stage with the advancement of gestation, although the maturation stage of megakaryocytes in the fetal bone marrow did not change with the advancement of gestation from 4 to 7 months, the megakaryocyte in the fetal bone marrow was less mature展开更多
Microphase separation and lamellar structure of segmented polyether- and polyester-polyurethanes have been investigated by means of transmission electron microscopy with the ruthenium tetroxide staining technique. The...Microphase separation and lamellar structure of segmented polyether- and polyester-polyurethanes have been investigated by means of transmission electron microscopy with the ruthenium tetroxide staining technique. The results show that the RuO_4 staining technique is simpler and may give better image contrast than other staining methods for this polymer. Microphase separation and lamellar structure of segmented polyether-and polyester-polyurethanes were directly observed and discussed.展开更多
A new complex Tb(IB)_3(phen)·2H_2O that gives off green fluorescence was synthesized. The results show that the strong fluorescence emitting of 490 and 545 nm for Tb 3+ in the Tb(IB)_3(phen)·2H_2O complex is...A new complex Tb(IB)_3(phen)·2H_2O that gives off green fluorescence was synthesized. The results show that the strong fluorescence emitting of 490 and 545 nm for Tb 3+ in the Tb(IB)_3(phen)·2H_2O complex is related to the transitions 5D_4-7F_6 and 5D_4-7F_5, respectively. The results indicate that the complex is concentrated on the nuclei regions in the section of onion toot tip tissues and the nuclei are high lighted under fluorescent microscope. It is possible that the Tb(IB)_3(phen)·2H_2O complex can be developed as a fluorescent dye in biological and pathological studies.展开更多
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) Immunofluorescence 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 neovascularization 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.
文摘In this study, mitotic metaphase chromosomes in mouse were identified by a new chromosome fluorescence banding technique combining DAPI staining with image analysis. Clear 4', 6-diamidino-2-phenylindole (DAPI) multiple bands like (J-hands could be produced in mouse. The Meta- Morph software was then used to generate linescans of pixel intensity for the banded chromosomes from short arm to long arm. These linescans were sufficient not only to identify each individual chromosome but also analyze the physical sites of bands in chromosome. Based on the results, the clear and accurate karyotype of mouse metaphase chromosomes was established. The technique is therefore considered to he a new method for cytological studies of mouse.
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.
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.
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supported by the program of The Project Supported by Natural Science Basic Research Plan in Shaanxi Province of China(No.SJ08-ZD05)
文摘Objective:The recent advent of flow cytometry(FCM),coupled with fluorescent dyes,has been successfully applied to assess mitochondrial function.The aim of this study was to investigate the feasibility and clinical significance of detecting sperm mitochondrial function and to evaluate sperm mitochondrial function by using Rhodamine 123/propidium(Rh123/PI)dual fluorescent staining and FCM in asthenospermia and oligoasthenozoospermia.Methods:Twenty-five fertile men(with normal sperm parameters)and 230 infertile patients were examined.Fifty-five patients of the above 230 patients were selected for idiopathic infertility samples and were divided into two groups:asthenospermia(n=30)and oligoasthenozoospermia(n=25).Rh123/PI dual fluorescent staining and FCM were carried out to examine sperm mitochondrial function.Results:Significant differences were found between the normal and abnormal semen samples(P0.05)when Rh123+/PI-,Rh123-/PI+and Rh123-/PI-sperm were examined by FCM,but there was no significant difference between the asthenospermia(P=0.469) and oligoasthenozoospermia group(P=0.950)when Rh123+/PI-and Rh123-/PI+sperm were then examined;however,a significant difference was found between the 2 groups(P=0.003)when Rh123-/PI-sperm were examined.There was no correlation between Rh123-/PI-sperm and semen parameters in the normal group,but there was a significant negative correlation between the sperm concentration and Rh123-/PI-sperm in asthenospermia and oligoasthenozoospermia patients(r=-0.509,-0.660;P=0.018,0.038).Conclusion:Rh123/PI dual fluorescent staining and FCM can provide reliable information to assess the quality of sperm and reveal differences in mitochondrial membrane potential in asthenospermia and oligoasthenozoospermia.
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.
文摘Microbial activity in the water injection system in oil and gas industry leads to an array of challenges, including biofouling, injectivity loss, reservoir plugging, and microbiologically influenced corrosion (MIC). An effective mitigation strategy requires online and real-time monitoring of microbial activity and growth in the system so that the operators can apply and adjust counter-measures quickly and properly. The previous study [1] identified DNA staining technology-with PicoGreen and SYBR Green dyes—as a very promising method for automated, online determination of microbial cell abundance in the vast Saudi Aramco injection seawater systems. This study evaluated DNA staining technology on detection limit, automation potential, and temperature stability for the construction of automated sensor prototype. DNA staining with SYBR Green dye was determined to be better suited for online and real-time monitoring of microbial activity in the Saudi Aramco seawater systems. SYBR Green staining does not require sample pre-treatment, and the fluorescence signal intensity is more stable at elevated temperatures up to 30℃. The lower detection limit of 2 × 10<sup>3</sup>/ml was achieved under the optimized conditions, which is sufficient to detect microbial numbers in Saudi Aramco injection seawater. Finally, the requirements for design and construction of SYBR-based automated sensor prototype were determined.
文摘Using potted seedlings of Chlorophytum comosum var. variegatum as the experimental materials, the effect of 2.0 mmol/L methyl orange ( Treatment T1 ), 1.0 mmol/L methyl violet ( Treatment T2 ) and 1.0 mmol/L neutral red ( Treatment T3 ) on the biomass, root-shoot ratio, leaf color indices, plant carbon and nitrogen nutrition were studied. The results showed that the biomass of Treatment T3 was significantly greater than that of treatments T1 and CK. The root-shoot ratio decreased significantly in treatments T1, T2 and T3 , and the decrease in T3 was most obvious. In all the three treatments with coloring agent, a ^* , b ^* and L ^* values were increased gradually, C value were decreased, H0 and CIRG were increased, and the leaves were pink. In addition, the contents of chlorophyll a, chlorophyll b, chlorophyll a + b and carotenoid were significantly decreased. The contents of soluble sugar and starch were also decreased, and the decrease in Treatment T2 was most significant. The contents of soluble protein and total nitrogen were increased, and the increase was most dramatic in Treatment T3. The carbon to nitrogen ratio was decreased. The results proved that staining can improve the ornamental value of indoor plants, despite its effects on plant carbon and nitrogen nutrition of C. comosum vat. variegatum, dyeing.
文摘A detailed morphological study of neurons in healthy and pathological conditions requires reasonably a number of special techniques, which may visualize the majority of neu- rons in a thick three-dimensional arrangement. A detailed visualization of neurons must include the cell body, most of the dendritic arbor, the dendritic spines, the axon, the axonal collaterals and the synapses. An ideal morphological technique for the study of degeneration and regeneration processes of the central nervous system must also visualize clearly the long and short neuronal circuits, as well as the dendritic and axonal bands and tracks.
基金Acknowledgments This study was supported by grants from the China National Natural Science Foundation (Nos. 30430530 and 30571337) and from the Momentous Research Project of the China Ministry of Science and Technology (No. 2006CB944003).
文摘The solution chemical and optical characteristics of formation of amine-terminated polyamidoamine dendrimer G2.0(NH2-PAMAM G2.0)-Au nanocomposites in the aqueous solution of NH2-PAMAM G2.0 at various mole ratios of Au(Ⅲ) to NH2-PAMAM G2.0 were studied by both UV-visible spectrometry and fluorospectrometry. The NH2-PAMAM G2.0-Au nanocomposites, with a type of structure in which one Au nanoparticle is surrounded by several NH2-PAMAM G2.0 dendrimers, emit strong bluish violet fluorescence, and are uniform, water soluble and biocompatible as well as very stable in frozen conditions. The size of gold nanoparticles in the nanocomposites is about 2.5 nm and decreases with the increase of NH2-PAMAM G2.0 concentration. The NH2-PAMAM G2.0 plays an important role in acting as host or micro-reactor for Au(Ⅲ) before Au(Ⅲ) reduction and acting as dispersant and stabilizer for gold nanoparticles after Au(Ⅲ) reduction. Preliminary experiments of cells staining to human embryonic lung fibroblast cell lines show that the NH2-PAMAM G2.0-Au nanocomposites can be used as optical imaging markers for bioanalyses and medical diagnoses.
文摘AIM: To evaluate clinicopathologic parameters and the clinical significance related lymphovascular invasion (LVI) by immunohistochemical staining (IHCS) in endoscopic submucosal dissection (ESD). METHODS: Between May 2005 and May 2010, a total of 348 lesions from 321 patients (mean age 63 ± 10 years, men 74.6%) with early gastric cancer (EGC) who met indication criteria after ESD were analyzed retrospectively. The 348 lesions were divided into the absolute (n = 100, differentiated mucosal cancer without ulcer ≤ 20 mm) and expanded (n = 248) indica-tion groups after ESD. The 248 lesions were divided into four subgroups according to the expanded ESD indication. The presence of LVI was determined by factor Ⅷ-related antigen and D2-40 assessment. We compared LVI IHCS-negative group with LVI IHCSpositive in each group. RESULTS: LVI by hematoxylin-eosin staining (HES) and IHCS were all negative in the absolute group, while was observed in only the expanded groups. The positive rate of LVI by IHCS was higher than that of LVI by HES (n = 1, 0.4% vs n = 11, 4.4%, P = 0.044). LVI IHCS-positivity was observed when the cancer invaded to the mucosa 3 (M3) or submucosa 1 (SM1) levels, with a predominance of 63.6% in the subgroup that included only SM1 cancer (P < 0.01). In a univariate analysis, M3 or SM1 invasion by the tumor was significantly associated with a higher rate of LVI by IHCS, but no factor was significant in a multivariate analysis. There were no cases of tumor recurrence or metastasis during the median 26 mo follow-up. CONCLUSION: EGCs of the absolute group are immunohistochemically stable. The presence of LVI may be carefully examined by IHCS in an ESD expanded indication group with an invasion depth of M3 or greater.
文摘Fast Plant(Brassica rapa,Cruciferae)leaf tissuefixed in glutaraldehyde-acrolein and post-fixed in os-mium,was examined for response to several easily-prepared heavy metal stains.Lead and uranium,separately and in combination,gave typical resultsacross the spectrum of cell organellets.As a single stainfollowing osmium,bismuth produced images seeminglyequivalent to lead and uranium.Phosphotungstic acidproduced very good membrane delineation but produceda washed-out background image similar to that from leadstaining.Carbohydrate compounds were especiallyresponsive to ruthenium;the cytoplasm and the matrixof all organelles were also stained very well.Theprocedures were no more demanding than traditionalstaining methods and may be easily used in research andteaching.Fast Plant materials are a reliable,quick andeasy source of living material.
文摘EST isozymes are one of the frequently used biochemical markers in genetic analysis of fungi. and the staining is an important process in electrophoresis analysis of studying Est.The effects were compared ainong different stain recipes for Est of 3 kinds of fungi-Lentinus edodes. Pleurotus sapidus. Phellinus igriarius and 2 kinds of plants-Populus sp and Brassica chinensis. Of the four kinds of Est staining recipes tested.the recipe α-acetic acid-naphther showed the best effect.and followed by β-aceticacid-naphther, semicontent α-aceticacid-naphther and α+β-aceticacidnathpher.
文摘AIM: To investigate whether performing immuno-histochemical CD3 staining, in order to improve the detection of intra-epithelial lymphocytosis, has an additional value in the histological diagnosis of celiac disease.METHODS: Biopsies obtained from 159 children were stained by hematoxylin and eosin(HE) and evaluated using the Marsh classification. CD3 staining was subsequently evaluated separately and independently.RESULTS: Differences in evaluation between the routine HE sections and CD3 staining were present in 20(12.6%) cases. In 10(6.3%) patients the diagnosis of celiac disease(Marsh Ⅱ and Ⅲ) changed on examination of CD3 staining: in 9 cases, celiac disease had initially been missed on the HE sections, while 1 patient had been over-diagnosed on the routine sections. In all patients, the final diagnosis based on CD3 staining, was concordant with serological results, which was not found previously. In the other 10(12.3%) patients, the detection of sole intra-epithelial lymphocytosis(Marsh Ⅰ) improved. Nine patients were found to have Marsh Ⅰ on CD3 sections, which had been missed on routine sections. Interestingly, the only patient with negative serology had Giardiasis. Finally, in 1 patient with negative serology, in whom Marsh Ⅰ was suspected on HE sections, this diagnosis was withdrawn after evaluation of the CD3 sections.CONCLUSION: Staining for CD3 has an additional value in the histological detection of celiac disease lesions, and CD3 staining should be performed when there is a discrepancy between serology and the diagnosis made on HE sections.
基金the National Natural Science Foundation of China, No. 90307013the Natural Science Foundation for Universities in Jiangsu Province, No. 05KJB180105a grant from Social Development Fund of Nantong City, No. S40052
文摘BACKGROUND: There are many methods for myelin staining, mordant, or oil-soluble dye or the special reaction of osmic acid with lipoid is used according to different principles. The commonly used methods are classic Well staining, classic lithium carbonate-haematine staining, fast green staining, silver staining, etc. Luxol Fast Blue can brightly stain myelin sheath, and has certain specificity. The background can be very clean if there is proper differentiation, whereas Luxol Fast Blue is cheap and convenient to operate, thus it is an ideal staining reagent for routine myelin sheath. OBJECTIVE: To show the corticospinal tract of normal adult rats with Luxol Fast Blue staining method. DESIGN: A repetitive measurement design. SETTINGS: Institute of Nuerobiology, Nantong University; Department of Rehabilitation Medicine, Affiliated Hospital of Nantong University. MATERIALS: Six healthy adult male SD rats of clean degree, weighing averagely 300 g, were provided by the experimental animal center of Nantong University. 1 g/L Luxol Fast Blue solution was provided by Sigma Company; Leica CM1900 cryostat microtome by Leica Company; Leica DMR microscope by Leica Company. METHODS: The experiment was carried out in the Staff Room of Human Anatomy, Nantong University in May 2005. The rats were given intraperitoneal injection of combined anesthetic (2 mL/kg), then the chest was open for perfusing saline and phosphate buffer containing formamint via heart. Brain and spinal cord were removed after 1 hour then fixed, then changed to phosphate buffer (pH 7.4) containing 300 g/L saccharu at 4 ℃, and stayed overnight, tissue blocks at pyramid, decussation of pyramid and cervical, thoracic, lumbar and sacral segments of spinal cord were removed to prepare continuous horizontal frozen sections (30 μm) after sedimentation, the sections were dried at room temperature. The corticospinal tract of normal adult rats were shown with Luxol Fast Blue staining method, and observed under Leica DMR microscope. MAIN OUTCOME MEASURES: Positive fibers in Luxol Fast Blue staining. RESULTS: After the Luxol Fast Blue staining, the labeled myelinated nerve fibers were bright blue. They located in the pyramid, decussation of pyramid and the ventral part of posterior funiculus in cervical, thoracic, lumbar and sacral segments of spinal cord. CONCLUSION: Luxol Fast Blue staining method may manifest the distribution of corticospinal tract with clear distinct in adult rats.
文摘Fetal liver tissues obtained from 28 human fetuses with gestation age from 3 to 6 months and fetal bone marrow from 35 human fetuses from 3 to 7 months were observed by immunochemical staining with anti-platelet GPⅡ b / Ⅲa monoclonal antibody and ABC technique. In the fetal liver, megakaryocytes were wholly located among growing fetal liver cells and near foci of hemopoiesis. Some megakaryocytes in the fetal liver were small7890- lymphoid-like megakaryocytes. The size of megakaryocytes both in the fetal liver (14.79 ± 4.52μm) and in the fetal bone marrow (16.08±7.39 μm) was small, which did not vary significantly over the gestation age ranging from 3 to 6 or 7 months. However, the maturation stage of megakaryocytes in the fetal liver shifted to more mature stage with the advancement of gestation, although the maturation stage of megakaryocytes in the fetal bone marrow did not change with the advancement of gestation from 4 to 7 months, the megakaryocyte in the fetal bone marrow was less mature
文摘Microphase separation and lamellar structure of segmented polyether- and polyester-polyurethanes have been investigated by means of transmission electron microscopy with the ruthenium tetroxide staining technique. The results show that the RuO_4 staining technique is simpler and may give better image contrast than other staining methods for this polymer. Microphase separation and lamellar structure of segmented polyether-and polyester-polyurethanes were directly observed and discussed.
文摘A new complex Tb(IB)_3(phen)·2H_2O that gives off green fluorescence was synthesized. The results show that the strong fluorescence emitting of 490 and 545 nm for Tb 3+ in the Tb(IB)_3(phen)·2H_2O complex is related to the transitions 5D_4-7F_6 and 5D_4-7F_5, respectively. The results indicate that the complex is concentrated on the nuclei regions in the section of onion toot tip tissues and the nuclei are high lighted under fluorescent microscope. It is possible that the Tb(IB)_3(phen)·2H_2O complex can be developed as a fluorescent dye in biological and pathological studies.