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Method for Combined Observation of Serial Sections of Stented Arteries Embedded in Resin by Light Microscopy and Transmission Electron Microscopy
摘要： We have developed a new method for obtaining information on whole tissues by light microscopy (LM) and ultrastructural features by transmission electron microscopy (TEM). This method uses serial sections of a stented artery embedded in resin. Stents were implanted in porcine coronary arteries in this study. The heart was perfusion fixed in a 2% paraformaldehyde and 1.25% glutaraldehyde mixed solution. The stented artery was then removed, fixed in 1% osmium, embedded in Quetol 651 resin, and sectioned serially. For LM, the black color of osmium was removed from the section by immersion in periodic acid and hydrogen peroxide after deplasticization. These sections were stained with hematoxylin and eosin and Elastica–Masson trichrome stain. For TEM, thin sections were re-embedded in Quetol 812 resin by the resupinate method and cut into ultrathin sections. A clear, fine structure was obtained, and organelles, microvilli, and cell junctions in the endothelium were easily observed. The combined observation of adjacent specimens by LM and TEM enabled us to relate histopathological changes in the millimeter scale to those in the nanometer scale.
关键词： porcine,stent,resupinate method,coronary artery,epoxy resin,transmission electron microscopy,light microscopy
更新于2025-09-23 15:22:29
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Light-microscopy methods in C. elegans research
摘要： Ever since Caenorhabditis elegans was introduced as a model system it has been tightly linked to microscopy, which has led to significant advances in understanding biology over the last decades. Developing new technologies therefore is an essential part in the endeavor to gain further mechanistic insights into developmental biology. This review will discuss state-of-the-art developments in quantitative light microscopy in the context of C. elegans research as well as the impact these technologies have on the field. We will highlight future developments that currently promise to revolutionize biological research by combining sequencing-based single-cell technologies with high-resolution quantitative imaging.
关键词： quantitative imaging,light microscopy,developmental biology,single-cell technologies,C. elegans
更新于2025-09-23 15:21:21
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Simplified Dynabeads Method Using Light Microscopy for Enumerating TCD4+ Lymphocytes in Resource-Limited Settings
摘要： Background: We demonstrated feasibility of implanting the Dynabeads method for CD4+ T lymphocyte enumeration in resource-poor settings (ANRS 1226 study). However, as this technique requires a fluorescence microscope which is not usually available in these settings, WHO has encouraged to simplify the method allowing TCD4+ lymphocyte counting under a light microscope. Methods: TCD4+ lymphocytes enumeration was assessed using Dynabeads after staining cells nuclei with non-fluorescent dyes and readings under light microscope (DLM). A total of 305 triple of values of CD4 cells counts were generated by both Dynabeads method using a light microscopy (DLM), Dynabeads method using a fluorescent microscope (DFM) and the single-platform flow cytometry technique (FCM). The accuracy of DLM was analyzed using 4 fresh blood samples showing 200, 400, 500 and 1000 cells/μl in FCM respectively. Correlations have been studied between the 3 methods. The DLM was then evaluated for its ability to correctly segregate absolute TCD4+ lymphocyte values at the thresholds of 200 cells/μl and 350 cells/μl. Findings: Cells nuclei staining with Sternheimer-Malbin, Turck1, and Giemsa allows TCD4+ lymphocytes enumeration using DLM. FCM has shown the greatest standard deviations and amplitudes. The reproducibility of DLM was better than FCM. The correlation coefficient between FCM and DFM was 0.975 and it was 0.973, 0.972 and 0.969 with DLM using Sternheimer-Malbin, Turck1 and Giemsa, respectively. The ability of DLM to correctly segregate TCD4+ lymphocyte values at the threshold of 200 cells/μl and 350 cells/μl was good. Conclusion: Reliable TCD4+ enumeration can be obtained with DLM. These results will contribute in resource-limited-settings to further reduce the cost of TCD4+ lymphocytes counting and make it more widely available in peripheral laboratories and even in central laboratories that face problems with maintenance and stock-out of reagents for flow cytometers.
关键词： Dynabeads method,Flow Cytometry,Resource-limited settings,Fluorescence microscopy,Light microscopy,TCD4+ lymphocytes enumeration
更新于2025-09-11 14:15:04
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[Methods in Molecular Biology] Barley Volume 1900 (Methods and Protocols) || Preparation of Barley Roots for Histological, Structural, and Immunolocalization Studies Using Light and Electron Microscopy
摘要： Microscopic investigations of biological objects are an integral part in plant research and most ?elds of life sciences. They allow the description of morphological, histological, and structural aspects of individual cells or tissues. Based on various cell biological tools and methods it is possible to characterize different plant genotypes or study their adaptation to changing environmental conditions. In combination with antibodies raised against speci?c antigens and epitopes light and electron microscopy enable investigation of the function of single genes/proteins in plant growth and development or their role related to abiotic or biotic stresses. Here, we describe sample preparation of barley roots for cell biological investigations using light and electron microscopy, to characterize morphological, structural, and functional aspects on root sections and the root surface.
关键词： Light microscopy,Electron microscopy,Fluorescence microscopy,Immunolocalization,Histology,Immunogold labeling
更新于2025-09-10 09:29:36
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Patenting an Accessory for a Light Microscope
摘要： The U.S. Patent and Trademark Office (PTO) encourages independent inventors who are not patent professionals to file and prosecute their own patent applications. Self-help books are useful guides for navigating through the PTO’s many laws and rules. When an inventor has an idea, he should document it, have the document witnessed, search all public records for novelty and unobviousness, and finally file an application with the PTO. The inventor defends his idea with an Examiner from the PTO and, if all requirements are met, receives a patent that grants the inventor a monopoly to make, use, and sell his invention for a period of time.
关键词： electrically variable focus lens,U.S. Patent and Trademark Office,light microscopy,patent applications,Inventions
更新于2025-09-10 09:29:36