We have developed a method where, after glutaraldehyde fixation, human hair shafts and insect cuticles are treated with ammonium thioglycolate (ATG) to improve ultrastructural staining. Conventional transmission elect...We have developed a method where, after glutaraldehyde fixation, human hair shafts and insect cuticles are treated with ammonium thioglycolate (ATG) to improve ultrastructural staining. Conventional transmission electron microscopic (TEM) preparations do not distinguish the A-layer and the exocuticles of hair shafts. However, after ATG treatment, the A-layer appears in higher contrast. ATG treatment has also been used to observe the fibrillar structure in the cortex. In the cuticle of beetles, the epicuticle is stained by ATG. Although the human hair shaft (keratin) and insect cuticle (chitin) are composed of different materials, both can be reduced by the ATG solution. The ammonium in the ATG solution reacts with hair shafts and insect cuticles, causing a reduction of swelled cuticles. Therefore, ATG not only stains, but also reduces human hair shafts and the cuticles of beetles.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
Uncoupling protein 1 (UCP1) expressed by the brown adipose tissue (BAT) in the mitochondrial crista acts as a homeostatic thermogenerator of eutherians. The evaluation of UCP1 expression in the BAT offers significant ...Uncoupling protein 1 (UCP1) expressed by the brown adipose tissue (BAT) in the mitochondrial crista acts as a homeostatic thermogenerator of eutherians. The evaluation of UCP1 expression in the BAT offers significant scientific insight, especially in studies targeting limited areas such as the periarterial and pericardial regions of small experimental mammals. However, the negligible amount of this adipose tissue would render the general quantitative evaluation of the protein unreliable because of lipid contamination and low protein concentration. To address this problem, we quantitatively evaluated UCP1 expression in the mitochondrion of the mouse interscapular BAT using immunoelectron microscopy and immunohistochemical studies using a combination of primary and secondary antibodies in scheme A (rabbit anti-UCP1 IgG/gold particle-conjugated goat anti-rabbit IgG), B (rabbit IgG/gold particle-conjugated goat anti-rabbit IgG), C (rabbit anti-UCP1 IgG/gold particle-unconjugated goat anti-rabbit IgG), and D (rabbit IgG/gold particle-unconjugated goat anti-rabbit IgG). Scheme A shows the immunopositive reaction of obvious gold particles in the mitochondrial area, whereas other procedures revealed less distinctive reactions. The distinctive gold particle immunoreaction comprised electrical high-density spots with a mean diameter of >5 nm. However, in scheme B, the electrical high-density spots were scattered outside the mitochondrion and were significantly smaller than 4 nm;schemes C and D demonstrated few immunoreactions. Logistic regression analysis between schemes A and B showed that the threshold diameter of the electrical high-density spots measuring >5 nm indicated a true positive immunoreaction to anti-UCP1 antibody specifically in the mitochondrial area. Minor statistical difference was observed in the primary anti-UCP1 antibody between polyclonal IgG and monoclonal antibodies. Therefore, immunoelectron microscopy might be useful for evaluating negligible protein expression in some limited areas, such as UCP1 expression in the BAT of small experimental animals.展开更多
The Turin Shroud, recently accessible for hands-on scientific research, is now extensively investigated. Its pinkish red blood stains that seem anomalous ones are studied by modern techniques (notably by resolute opti...The Turin Shroud, recently accessible for hands-on scientific research, is now extensively investigated. Its pinkish red blood stains that seem anomalous ones are studied by modern techniques (notably by resolute optical microscopy and scanning electron microscopy coupled with energy dispersive X-ray). Exploration by these techniques of a blood stain located on the face permits us to discover some red-colour particles (hematite, biotite and cinnabar) of exogenous material in this stain. We finally characterize these red-colour particles and try to explain their presences in the blood stain. Globally, all these red-colour particles cannot explain all of the reddish appearance of the area under study.展开更多
文摘We have developed a method where, after glutaraldehyde fixation, human hair shafts and insect cuticles are treated with ammonium thioglycolate (ATG) to improve ultrastructural staining. Conventional transmission electron microscopic (TEM) preparations do not distinguish the A-layer and the exocuticles of hair shafts. However, after ATG treatment, the A-layer appears in higher contrast. ATG treatment has also been used to observe the fibrillar structure in the cortex. In the cuticle of beetles, the epicuticle is stained by ATG. Although the human hair shaft (keratin) and insect cuticle (chitin) are composed of different materials, both can be reduced by the ATG solution. The ammonium in the ATG solution reacts with hair shafts and insect cuticles, causing a reduction of swelled cuticles. Therefore, ATG not only stains, but also reduces human hair shafts and the cuticles of beetles.
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Uncoupling protein 1 (UCP1) expressed by the brown adipose tissue (BAT) in the mitochondrial crista acts as a homeostatic thermogenerator of eutherians. The evaluation of UCP1 expression in the BAT offers significant scientific insight, especially in studies targeting limited areas such as the periarterial and pericardial regions of small experimental mammals. However, the negligible amount of this adipose tissue would render the general quantitative evaluation of the protein unreliable because of lipid contamination and low protein concentration. To address this problem, we quantitatively evaluated UCP1 expression in the mitochondrion of the mouse interscapular BAT using immunoelectron microscopy and immunohistochemical studies using a combination of primary and secondary antibodies in scheme A (rabbit anti-UCP1 IgG/gold particle-conjugated goat anti-rabbit IgG), B (rabbit IgG/gold particle-conjugated goat anti-rabbit IgG), C (rabbit anti-UCP1 IgG/gold particle-unconjugated goat anti-rabbit IgG), and D (rabbit IgG/gold particle-unconjugated goat anti-rabbit IgG). Scheme A shows the immunopositive reaction of obvious gold particles in the mitochondrial area, whereas other procedures revealed less distinctive reactions. The distinctive gold particle immunoreaction comprised electrical high-density spots with a mean diameter of >5 nm. However, in scheme B, the electrical high-density spots were scattered outside the mitochondrion and were significantly smaller than 4 nm;schemes C and D demonstrated few immunoreactions. Logistic regression analysis between schemes A and B showed that the threshold diameter of the electrical high-density spots measuring >5 nm indicated a true positive immunoreaction to anti-UCP1 antibody specifically in the mitochondrial area. Minor statistical difference was observed in the primary anti-UCP1 antibody between polyclonal IgG and monoclonal antibodies. Therefore, immunoelectron microscopy might be useful for evaluating negligible protein expression in some limited areas, such as UCP1 expression in the BAT of small experimental animals.
文摘The Turin Shroud, recently accessible for hands-on scientific research, is now extensively investigated. Its pinkish red blood stains that seem anomalous ones are studied by modern techniques (notably by resolute optical microscopy and scanning electron microscopy coupled with energy dispersive X-ray). Exploration by these techniques of a blood stain located on the face permits us to discover some red-colour particles (hematite, biotite and cinnabar) of exogenous material in this stain. We finally characterize these red-colour particles and try to explain their presences in the blood stain. Globally, all these red-colour particles cannot explain all of the reddish appearance of the area under study.
