Fluorochrome Staining
Fluorochrome Staining - Related Videos
07:29
Undecalcified Bone Preparation for Histology, Histomorphometry and Fluorochrome Analysis
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Cited by 32 •
2010
Undecalcified bone histology provides important information for a variety of clinical and research applications. It is technically challenging, particularly with large size specimens. This video illustrates the process of producing good quality sections and demonstrates the technical difficulties and methods with which to overcome them.
10:58
Discrimination of Seven Immune Cell Subsets by Two-fluorochrome Flow Cytometry
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2019
Here, we present a flow cytometric protocol to identify CD4+ and CD8+ T cells, γδ T cells, B cells, NK cells and monocytes in human peripheral blood by using only two fluorochromes instead of seven. With this approach, five additional markers can be recorded on most flow cytometers.
03:18
Flow Cytometry Staining of Mycobacterium tuberculosis Infected Macrophage Subpopulations
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2025
This video showcases a flow cytometric gating strategy utilizing a multi-color panel to analyze cell viability and specific fluorescent-conjugated anti-human antibody markers. This approach makes it possible to determine the percentage of Mycobacterium tuberculosis-infected cell populations accurately.
11:33
Microscopy and Staining: Gram, Capsule, and Endospore Staining
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2023
Source: Rhiannon M. LeVeque1, Natalia Martin1, Andrew J. Van Alst1, and Victor J. DiRita1 1 Department of Microbiology and Molecular Genetics, Michigan State University, East Lansing, Michigan, United States of America Bacteria are diverse microorganisms found nearly everywhere on Earth. Many properties help distinguish them from each other, including but not limited to Gram-staining type, shape and arrangement, production of capsule, and formation of spores. To observe these properties, one...
03:12
May-Grunwald Giemsa Staining: A Method to Stain Bone Marrow Cells
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2023
This video describes the technique of staining bone marrow cells using May- Grünwald Giemsa Staining. In the protocol, we will stain the murine myeloid leukemia cells, C149 cells, to analyze the morphology and differential counting.
FAQs
A fluorochrome absorbs light at a defined excitation wavelength and then emits light at a longer wavelength. This separation allows suitable microscopy systems to distinguish the emitted signal from the illumination used to excite it. Matching the fluorochrome’s light response with appropriate imaging settings helps reveal labeled structures clearly and supports more interpretable biological observations.
Signal localization depends on how the fluorochrome associates with the sample. Selective binding can target particular cellular structures, antibody conjugation can mark proteins or other recognized molecules, and nucleic-acid association can highlight gene-related features. These alternatives provide different routes to spatial information, so the chosen labeling mechanism should match the biological feature being investigated.
Staining conditions influence whether labeling remains sufficiently selective within a biological sample, while imaging settings determine how reliably the resulting fluorescence is detected and interpreted. Poor control of either stage can make cellular features harder to distinguish or complicate conclusions about molecular distribution. Careful optimization therefore supports clearer morphology, localization, and comparative observations.
A typical workflow includes labeling the relevant cellular structure or molecule, exposing the sample to light that excites the fluorochrome, and observing the longer-wavelength emitted signal with suitable microscopy. Depending on the experiment, the labeled sample may then be assessed for morphology, protein distribution, viability, or gene-related features. Consistent staining and imaging conditions improve interpretability.
Fluorochrome staining supports several complementary approaches, including fluorescence microscopy, immunofluorescence, flow cytometry, and live-cell analysis. Microscopy can show spatial organization, immunofluorescence can reveal protein distribution through antibody-based labeling, flow cytometry can evaluate fluorescent properties across cells, and live-cell analysis can examine fluorescence-associated features in living biological material.
The method can make cell morphology, protein distribution, viability, and gene-related features observable through fluorescent signals. Its value lies in connecting a label with a specific cellular or molecular location, rather than simply showing that fluorescence is present. In research and diagnostic studies, these observations can help characterize samples and examine how biological features are distributed.