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
Telomerase is a ribonucleoprotein enzyme, which synthesizes telomeric repeats (TTAGGG) n.While germline cells and most malignant tumor cells express telomerase activity, normal somatic cells aregenerally deficient in ...Telomerase is a ribonucleoprotein enzyme, which synthesizes telomeric repeats (TTAGGG) n.While germline cells and most malignant tumor cells express telomerase activity, normal somatic cells aregenerally deficient in telomerase activity. Our objective was to detect telomerase activity of human cells bysilver staining with telorneric repeat amplification protocol (TRAP) which is easy and quick. Comparing withradioisotopic TRAP, we examined the telomerase activity in telomerase-positive 293-cell and RNase-pretreated and heat-pretreated negative controls by silver staining TRAP. We detected telomerase activity in 2 strainsof human liver tumor cells (QGY7701 and SMMC7721 ). The 293 cells (only 10 cells) and the 2 strains ofhuman liver tumor cells were all positive. while telomerase activity was not detected in the negative controls.These data suggest that non-radioisotopic silver staining TRAP is a specific, sensitive and fast assay fortelomerase activity. It was verified that the 2 strains of human liver tumor cells express telomerase activity.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
Lung smears of mice and lung sections of rats or human case with Pneumocystis cariniiinfection were stained using the Grocott's modification method of Gomori's methenamine-silver nitratetechnic, in which 5% so...Lung smears of mice and lung sections of rats or human case with Pneumocystis cariniiinfection were stained using the Grocott's modification method of Gomori's methenamine-silver nitratetechnic, in which 5% sodium periodate and 5% chromic acid were used as oxidant respectively. Theoxidation time for the mouse lung smears was 5,15,60 minutes and the oxidation temperature was 20℃.The time of silver impregnation was 90 minutcs and the temperature was 60℃ for the all smearo. Whenthe oxidation time was under 15 minutes. Pneumocystis cariniic cysts showed light or dark brown, and theparenthesis-like structure could clearly be found in part of the cysts. However, if the time of oxidationWas longer, the cysts showed black and secmed to have damaged. In the same batch of the mouse lungsmears oxidated for 5 minutes, the samiples oxidated by sodium periodate showed more the cysts with theparen thesis-like structure than those oxidated by chromic acid.In the rat or patient's lung sectionsoxidated by. sodium periodate, this structure could also be found. The result of the experiment showsthat sodium periodate as an oxidant in the subsequent step of the the silver impregnation is preferable tochromic acid. And then,it is useful to clinical practice that the step of sodium bisulfate can be omittedin the study.展开更多
Silver Diamine Fluoride (SDF) is colorless and alkaline with a pH of 10. It has been used in Japan and other international countries for decades. The Food and Drug Administration gave approval for it as a means of tre...Silver Diamine Fluoride (SDF) is colorless and alkaline with a pH of 10. It has been used in Japan and other international countries for decades. The Food and Drug Administration gave approval for it as a means of treating hypersensitivity for individuals with chronic teeth pain. SDF is also used as a method to treat and arrest dental caries. SDF application is limited due to its negative esthetic effects, which is a black stain where the cavity was present on the tooth. Topical application of potassium iodide applied immediately after SDF has been shown in studies to reduce the color change caused by SDF. This study used topical application of silver diamine fluoride (SDF) and potassium iodide (KI) treatments on bovine teeth to determine if SDF and KI were efficacious in the treatment for carious lesions. The color change was detected by use of spectrophotometric analysis to determine L, a and b readings that demarcate light and color values following staining. The conclusion was made that the application of SDF followed directly by KI treatment produced L, a and b spectrophotometric values that indicated a significant reduction in teeth staining than the application of SDF alone. Therefore, this study supports the idea that SDF and KI can be used to treat carious lesions on bovine teeth while retaining surface enamel coloration.展开更多
in situ hybridization or hybridocytochemistry is a powerful tool for detection and localization of gene expression. It is also the most commonly used procedure to localize specific DNA or RND sequences in tissue secti...in situ hybridization or hybridocytochemistry is a powerful tool for detection and localization of gene expression. It is also the most commonly used procedure to localize specific DNA or RND sequences in tissue sections or cell preparations. Probes are usually展开更多
Antisera against abseisic-acid-binding proteins(Ab Ⅰ)and anti-idiotypic antibodies to ABAmonoclonal antibody(Ab Ⅱ)have been developed and tested to comparatively localize abacisic-acid-bindingproteins(ABBPs)in maize...Antisera against abseisic-acid-binding proteins(Ab Ⅰ)and anti-idiotypic antibodies to ABAmonoclonal antibody(Ab Ⅱ)have been developed and tested to comparatively localize abacisic-acid-bindingproteins(ABBPs)in maize root tips using immunocytochemistry technique.Based on the staining pattern ob-served along the root tip,an apicobasal gradient of ABBP has been revealed with either Ab Ⅰ or Ab Ⅱ.Thelabels are mainly concentrated in,the root cap and apical meristem.In the elongation zone and root hair zone,only cells in pericycle and epidermis are obviously labeled.The silver labels are distributed evenly in thewhole cell of the apical meristem,but in the cells of the root cap and elongation zone,the positive stainingdegree in the cytoplasm is reduced even to completely negative staining;nevertheless,the nuclei and plasmamembranes are strongly labeled.As in the labeled cells of the root hair zone,only some immunoactive sitescan be seen in the nucleus compacted to the edge of the cells.The staining pattern in the whole root tip alsoshows obvious dependence of the ABBPs localization on the activity of the nucleus.The significance ofABBPs localization is discussed.展开更多
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Telomerase is a ribonucleoprotein enzyme, which synthesizes telomeric repeats (TTAGGG) n.While germline cells and most malignant tumor cells express telomerase activity, normal somatic cells aregenerally deficient in telomerase activity. Our objective was to detect telomerase activity of human cells bysilver staining with telorneric repeat amplification protocol (TRAP) which is easy and quick. Comparing withradioisotopic TRAP, we examined the telomerase activity in telomerase-positive 293-cell and RNase-pretreated and heat-pretreated negative controls by silver staining TRAP. We detected telomerase activity in 2 strainsof human liver tumor cells (QGY7701 and SMMC7721 ). The 293 cells (only 10 cells) and the 2 strains ofhuman liver tumor cells were all positive. while telomerase activity was not detected in the negative controls.These data suggest that non-radioisotopic silver staining TRAP is a specific, sensitive and fast assay fortelomerase activity. It was verified that the 2 strains of human liver tumor cells express telomerase activity.
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Lung smears of mice and lung sections of rats or human case with Pneumocystis cariniiinfection were stained using the Grocott's modification method of Gomori's methenamine-silver nitratetechnic, in which 5% sodium periodate and 5% chromic acid were used as oxidant respectively. Theoxidation time for the mouse lung smears was 5,15,60 minutes and the oxidation temperature was 20℃.The time of silver impregnation was 90 minutcs and the temperature was 60℃ for the all smearo. Whenthe oxidation time was under 15 minutes. Pneumocystis cariniic cysts showed light or dark brown, and theparenthesis-like structure could clearly be found in part of the cysts. However, if the time of oxidationWas longer, the cysts showed black and secmed to have damaged. In the same batch of the mouse lungsmears oxidated for 5 minutes, the samiples oxidated by sodium periodate showed more the cysts with theparen thesis-like structure than those oxidated by chromic acid.In the rat or patient's lung sectionsoxidated by. sodium periodate, this structure could also be found. The result of the experiment showsthat sodium periodate as an oxidant in the subsequent step of the the silver impregnation is preferable tochromic acid. And then,it is useful to clinical practice that the step of sodium bisulfate can be omittedin the study.
文摘Silver Diamine Fluoride (SDF) is colorless and alkaline with a pH of 10. It has been used in Japan and other international countries for decades. The Food and Drug Administration gave approval for it as a means of treating hypersensitivity for individuals with chronic teeth pain. SDF is also used as a method to treat and arrest dental caries. SDF application is limited due to its negative esthetic effects, which is a black stain where the cavity was present on the tooth. Topical application of potassium iodide applied immediately after SDF has been shown in studies to reduce the color change caused by SDF. This study used topical application of silver diamine fluoride (SDF) and potassium iodide (KI) treatments on bovine teeth to determine if SDF and KI were efficacious in the treatment for carious lesions. The color change was detected by use of spectrophotometric analysis to determine L, a and b readings that demarcate light and color values following staining. The conclusion was made that the application of SDF followed directly by KI treatment produced L, a and b spectrophotometric values that indicated a significant reduction in teeth staining than the application of SDF alone. Therefore, this study supports the idea that SDF and KI can be used to treat carious lesions on bovine teeth while retaining surface enamel coloration.
文摘in situ hybridization or hybridocytochemistry is a powerful tool for detection and localization of gene expression. It is also the most commonly used procedure to localize specific DNA or RND sequences in tissue sections or cell preparations. Probes are usually
文摘Antisera against abseisic-acid-binding proteins(Ab Ⅰ)and anti-idiotypic antibodies to ABAmonoclonal antibody(Ab Ⅱ)have been developed and tested to comparatively localize abacisic-acid-bindingproteins(ABBPs)in maize root tips using immunocytochemistry technique.Based on the staining pattern ob-served along the root tip,an apicobasal gradient of ABBP has been revealed with either Ab Ⅰ or Ab Ⅱ.Thelabels are mainly concentrated in,the root cap and apical meristem.In the elongation zone and root hair zone,only cells in pericycle and epidermis are obviously labeled.The silver labels are distributed evenly in thewhole cell of the apical meristem,but in the cells of the root cap and elongation zone,the positive stainingdegree in the cytoplasm is reduced even to completely negative staining;nevertheless,the nuclei and plasmamembranes are strongly labeled.As in the labeled cells of the root hair zone,only some immunoactive sitescan be seen in the nucleus compacted to the edge of the cells.The staining pattern in the whole root tip alsoshows obvious dependence of the ABBPs localization on the activity of the nucleus.The significance ofABBPs localization is discussed.
