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FISH - Fluorescent In-situ Hybridization

FISH - Fluorescent In-situ Hybridization

Fluorescence in situ hybridization, or FISH, was developed in the early 1980s and has quickly become one of the most widely used techniques in cytogenetics. Labeled probes are used to bind complementary DNA or RNA sequences on a chromosome or in a region within a cell. Earlier, the probes could only be obtained by cloning or reverse transcription of a DNA template. Currently, the probe oligonucleotides can be synthesized synthetically. Additionally, with the advancement of optical techniques,...

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Related Experiment Video

Updated: Jun 27, 2025

Author Spotlight: Single-Molecule Surface-Enhanced Raman Scattering Measurements Enabled by Plasmonic DNA Origami Nanoantennas

Author Spotlight: Single-Molecule Surface-Enhanced Raman Scattering Measurements Enabled by Plasmonic DNA Origami Nanoantennas

Published on: July 21, 2023

High-Throughput, Label-Free Detection of DNA Origami in Single-Cell Suspensions Using origamiFISH-Flow.

Shana Alexander¹, Wendy Xueyi Wang^1,2, Chung-Yi Tseng¹

¹Institute of Biomedical Engineering, University of Toronto, Toronto, ON, M5S 3G9, Canada.

Small (Weinheim an Der Bergstrasse, Germany)

|April 30, 2024

Summary

This summary is machine-generated.

A new method, origamiFISH-Flow, enables high-throughput quantification of DNA nanostructures (DNs) in single cells. This technique improves detection sensitivity and allows detailed analysis of DN-cell interactions for bio-application design.

Keywords:

DNA origami delivery flow cytometry in situ hybridization nanomaterials

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Last Updated: Jun 27, 2025

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Single Cell Analysis Of Transcriptionally Active Alleles By Single Molecule FISH

Single Cell Analysis Of Transcriptionally Active Alleles By Single Molecule FISH

Published on: September 20, 2020

Area of Science:

Biotechnology
Nanotechnology
Cell Biology

Background:

Structural DNA nanotechnology allows for the creation of custom nanoscale devices with potential biological applications.
Understanding how DNA nanostructure (DN) design influences interactions with cells in vitro and in vivo is crucial but not well-characterized.
Existing methods for detecting DNs in cells, like fluorescent tags, have limitations in sensitivity and specificity.

Purpose of the Study:

To extend the origamiFISH technique for high-throughput quantification of DNs in single-cell suspensions using flow cytometry.
To develop a method, origamiFISH-Flow, capable of analyzing DN-cell interactions in various cell types, including nonadherent immune cells.
To provide a sensitive and quantitative tool for interrogating DN uptake and distribution in biological systems.

Main Methods:

Adaptation of the origamiFISH technique for flow cytometry analysis (origamiFISH-Flow).
High-throughput quantification of DNs in single-cell suspensions at rates of 10,000 cells per second.
Concurrent cell-type and cell-state characterization using immunostaining alongside DN detection.

Main Results:

origamiFISH-Flow demonstrated a 20-fold higher signal-to-noise ratio for DN detection compared to traditional dye labeling.
The method successfully identified >25-fold more DN-positive cells at low picomolar concentrations.
Validated profiling of DN uptake across diverse cell lines and splenocytes, and quantified in vivo DN accumulation in lymphoid organs.

Conclusions:

origamiFISH-Flow is a powerful, high-throughput tool for quantifying DNA nanostructures within single cells.
This method significantly enhances the sensitivity and scope of DN-cell interaction studies.
origamiFISH-Flow provides valuable insights for optimizing DNA nanostructure design in biomedical applications.