Fluorescence Light Microscopy
Fluorescence Light Microscopy - Related Videos
13:01
Using Light Sheet Fluorescence Microscopy to Image Zebrafish Eye Development
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Cited by 60 •
2016
Light sheet fluorescence microscopy is an excellent tool for imaging embryonic development. It allows recording of long time-lapse movies of live embryos in near physiological conditions. We demonstrate its application for imaging zebrafish eye development across wide spatio-temporal scales and present a pipeline for fusion and deconvolution of multiview datasets.
11:16
Correlative Light- and Electron Microscopy Using Quantum Dot Nanoparticles
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Cited by 12 •
2016
A method is described whereby quantum dot (QD) nanoparticles can be used for correlative immunocytochemical studies of epoxy embedded human pathology tissue. We employ commercial antibody fragment conjugated QDs that are visualized by widefield fluorescence light microscopy and transmission electron microscopy.
10:00
Low-Cost Cryo-Light Microscopy Stage Fabrication for Correlated Light/Electron Microscopy
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Cited by 11 •
2011
We demonstrate the fabrication of a low-cost cryogenic stage designed to fit most reflected light microscopes. This lab-built cryogenic stage enables efficient and reliable correlative imaging between cryo-light and cryo-electron microscopy.
06:41
Light Sheet Fluorescence Microscopy of Plant Roots Growing on the Surface of a Gel
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Cited by 24 •
2017
This protocol shows a plant sample preparation method for light-sheet microscopy. The setup is characterized by mounting the plant vertically on the surface of a gel and letting it grow in controlled bright conditions. This allows long-term observation of plant organ development in standardized conditions.
08:35
Introduction to Light Microscopy
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2023
The light microscope is an instrument used by researchers in many different fields to magnify specimens to as much as a thousand times their original size. In its simplest form, it is composed of a clear lens that magnifies the sample and a light source to illuminate it. However, most light microscopes are much more complex and house numerous fine-tuned lenses with tightly controlled dimensions all within the body of the microscope itself and in components such as the objectives and eyepieces.
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Optical filters separate the emitted fluorescent signal from the light used to illuminate the sample, while specialized objectives collect and focus that signal for observation. This separation helps labeled structures, cells, or microorganisms appear against a darker background. In infection and immunology studies, the resulting contrast supports clearer localization of markers, pathogens, and host proteins.
A fluorophore emits light at a longer wavelength than the light it absorbs. Fluorescence Light Microscopy uses this difference to distinguish emitted signal from the illumination that excites the molecule. The wavelength separation is therefore central to detecting labeled targets and producing contrast between fluorescently marked material and the surrounding sample.
Fluorescent antibodies connect optical detection with molecular recognition. By labeling immune-cell markers, pathogens, or host proteins, they allow investigators to determine where selected targets occur within a sample and to examine their spatial relationships. This makes antibody-based labeling useful for cell phenotyping, pathogen detection, and studying interactions between infectious agents and host cells.
The workflow begins by selecting a target and an appropriate fluorescent label, such as an antibody directed toward an immune-cell marker, pathogen, or host protein. Investigators then examine the labeled material with the required filters and objective. Using a fixed or living sample determines whether the analysis focuses on preserved material or observations made in living cells.
It is useful when researchers need to identify particular cell populations, detect microorganisms, or locate host and pathogen components within the same experimental setting. The method can reveal how targets are distributed and whether they occur in related cellular locations. These capabilities support immune-cell phenotyping, pathogen detection, and analysis of host-pathogen relationships.
Fluorescence Light Microscopy can show changes in cellular organization, the location of immune or infectious markers, and relationships between host proteins and pathogens. Comparing fluorescent patterns across samples can therefore provide evidence of altered organization or immune responses. Interpretation depends on which structures or molecules were labeled and where their signals appear within the sample.