Widefield Microscope
Widefield Microscope - Related Videos
09:45
Ex vivo Imaging of T Cells in Murine Lymph Node Slices with Widefield and Confocal Microscopes
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Cited by 21 •
2011
This protocol describes a method to image fluorescent T cells introduced into lymph node slices. The technique permits real-time analyses of T cell migration with traditional widefield fluorescence or confocal microscopes.
07:02
Stiffness Measurement of Soft Silicone Substrates for Mechanobiology Studies Using a Widefield Fluorescence Microscope
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Cited by 19 •
2018
Substrates with stiffness in the kilopascal-range are useful to study the response of cells to physiologically relevant micro-environment stiffness. Using just a widefield fluorescence microscope, the Young's modulus of soft silicone gels can be determined using an indentation with a suitable sphere.
04:57
Proper Care and Cleaning of the Microscope
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Cited by 2 •
2008
Keeping the microscope optics clean is important for high-quality imaging. Dust, fingerprints, excess immersion oil, or mounting medium on or in a microscope causes reduction in contrast and resolution. DIC is especially sensitive to contamination and scratches on the lens surfaces. This protocol details the procedure for keeping the microscope clean.
08:08
Major Components of the Light Microscope
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2008
The light microscope is a basic tool for the cell biologist, who should have a thorough understanding of how it works, how it should be aligned for different applications, and how it should be maintained as required to obtain maximum image-forming capacity and resolution. The components of the microscope are described in detail here.
09:33
Automated Slide Scanning and Segmentation in Fluorescently-labeled Tissues Using a Widefield High-content Analysis System
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Cited by 2 •
2018
Here we describe a protocol for the automated segmentation of fluorescently labeled tissues on slides using a widefield high-content analysis system (WHCAS). This protocol has wide-ranging applications in any field which involves the quantitation of fluorescent markers in biological tissues including the biological sciences, medical engineering, and health sciences.
FAQs
A fluorescence observation depends on coordinated illumination and detection paths. Excitation light reaches fluorophores in the specimen, and the resulting emitted light passes through optical filters and the objective before reaching a camera or eyepiece. This arrangement links the detected image to labeled structures, allowing investigators to examine protein localization or cellular organization across the field.
Optical filters determine how emitted fluorescence is handled before image formation. In the fluorescence pathway, light produced by fluorophores travels through these filters and then through the objective to the camera or eyepiece. Their position in the detection path helps preserve the fluorescence information used to interpret cellular or tissue patterns.
Transmitted-light and fluorescence modes provide different biological views. Transmitted-light imaging detects light that passes through or is scattered by the specimen, whereas fluorescence imaging follows light emitted by fluorophores after excitation. Researchers can select the mode according to whether they need general specimen structure or information associated with fluorescently marked features, such as protein localization.
A practical workflow begins by placing the specimen in the field and selecting transmitted-light or fluorescence illumination according to the observation goal. In fluorescence mode, excitation light passes through the sample, and emitted light travels through optical filters and the objective to a camera or eyepiece. The resulting images can support observation, screening, or quantitative analysis.
This instrument is useful when rapid observation across a broad field is needed. In biology, applications include routine viewing of cells, tissues, and microorganisms, screening specimens, teaching, and live-cell studies. Its relatively simple design supports accessible visual inspection while still allowing investigators to examine cellular structure, tissue organization, or fluorescently localized proteins.
Images from this instrument can reveal cellular structure, protein localization, and tissue organization, while also supporting quantitative analysis. The broad field of view is valuable for rapid screening and live-cell studies, and the relatively simple design suits teaching. These capabilities make the technique relevant to both routine biological observation and measurements derived from recorded images.