Hcr-fish
Hcr-fish - Related Videos
08:55
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07:25
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02:32
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2019
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07:41
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Cited by 2 •
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05:12
Swimming Performance Assessment in Fishes
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Cited by 55 •
2011
The lives of the majority of fish are predicated on swimming. This protocol describes techniques for capturing a range of swimming modes available to individual and schooling fish, and includes metrics associated with swimming physiology and behaviour.
FAQs
Target-specific probes first bind complementary nucleic acid sequences inside the sample. This binding creates a site that activates fluorophore-labeled hairpins. The hairpins alternately open and polymerize into a chain, concentrating many fluorescent labels at the original target location. Because assembly occurs through probe-triggered hairpin interactions, the signal is amplified without enzymatic amplification.
HCR-FISH generates amplified fluorescence through self-assembling hairpins rather than an enzyme-driven amplification reaction. This mechanism supports detection while the target remains in intact cells and tissues, allowing researchers to relate transcript-associated signals to neuronal structure, brain-region organization, and other anatomical features that might be difficult to interpret without preserved spatial context.
Researchers can use multiplexed probe sets to detect multiple transcripts in the same neural sample. Each target-specific probe set identifies a selected nucleic acid sequence, while the resulting fluorescent signals reveal where those targets occur relative to one another. This makes it possible to examine spatial relationships among transcripts rather than analyzing each gene-expression pattern in isolation.
The method preserves cellular and anatomical context while mapping nucleic acid signals across neurons, brain regions, and other neural tissues. Consequently, researchers can connect gene-expression patterns with the locations of cells and structures in the nervous system. This spatial organization helps interpret molecular differences in relation to neural development, cell identity, connectivity, or disease-associated changes.
A typical workflow begins with target-specific probes binding selected RNA or DNA sequences in intact cells or tissues. Fluorophore-labeled hairpins are then introduced so probe-bound sites trigger alternating hairpin opening and polymerization. The assembled fluorescent chains produce localized signals that can be imaged and compared across cells, anatomical regions, or multiplexed transcript targets.
HCR-FISH is useful when a study needs both gene-expression information and the position of that expression within neural tissue. Applications include mapping transcripts during neural development, distinguishing cell identities, examining connectivity-related molecular patterns, and investigating disease-associated changes. Its value is greatest when spatial relationships among cells, regions, or multiple transcripts are scientifically important.
Images can show the presence and localization of selected RNA or DNA sequences within intact neural samples. With multiplexed probe sets, investigators can compare transcript distributions and identify spatial relationships among molecular signals. These observations support analyses of gene expression across neurons and brain regions, helping connect molecular patterns with neural organization and disease-related changes.