hybridization based on the mechanism of the hybridization chain reaction (HCR) has addressed multi-decade challenges that impeded imaging of mRNA expression in diverse organisms, offering a unique combination of multiplexing, quantitation, sensitivity, resolution and versatility. Here, with third-generation HCR, we augment these capabilities using probes and amplifiers that combine to provide automatic background suppression throughout the protocol, ensuring that reagents will not generate amplified background even if they bind non-specifically within the sample. Automatic background suppression dramatically enhances performance and robustness, combining the benefits of a higher signal-to-background ratio with the convenience of using unoptimized probe sets for new targets and organisms. HCR v3.0 enables three multiplexed quantitative analysis modes: (1) qHCR imaging - analog mRNA relative quantitation with subcellular resolution in the anatomical context of whole-mount vertebrate embryos; (2) qHCR flow cytometry - analog mRNA relative quantitation for high-throughput expression profiling of mammalian and bacterial cells; and (3) dHCR imaging - digital mRNA absolute quantitation via single-molecule imaging in thick autofluorescent samples.
Download full-text PDF |
Source |
|---|---|
| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6031405 | PMC |
| http://dx.doi.org/10.1242/dev.165753 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
Cocatalysis of catalytic hairpin assembly and ternary heterojunction BiS@MoS@BiMoO promotes ultra-sensitive electrochemical of detection MiRNA-21.
Mikrochim Acta
January 2025
USST-UH International Joint Laboratory for Tumor Diagnosis and Energy Treatment, University of Shanghai for Science and Technology, Shanghai, 200093, China.
Ternary heterojunction BiS/MoS/BiMoO was designed as a signal probe to develop a dual signal amplification strategy empowered electrochemical biosensor for sensitive miRNA-21 detection by combining with catalytic hairpin assembly (CHA). The combination of the BiS/MoS/BiMoO heterojunction as a tracer indication probe and the CHA amplification strategy not only took fully use of the highly dense nanowire interwoven structure and superior active region of the probe, but also endowed the ability to improve the molecular hybridization efficiency by collision, which significantly avoided the cumbersome chain design and greatly simplified the step-by-step construction of the electrode surface. Hairpin H1 was first added dropwise to the gold nanoparticle-decorated electrode surface, and then opened by the introduced miRNA-21 to initiate the specific hybridization.
View Article and Find Full Text PDFsp. nov., sp. nov. and sp. nov. isolated from a cave, and an emended description of the genus .
Int J Syst Evol Microbiol
January 2025
Department of Biological Sciences and Biotechnology, Hannam University, Daejeon 34054, Republic of Korea.
Six Gram-reaction-positive, strictly aerobic, mycelium-forming actinobacteria were isolated from soils collected from a natural cave in Jeju, Republic of Korea. The isolates produced well-developed, branched, substrate mycelia and white aerial mycelia that differentiated into straight or flexuous chains of smooth-surfaced spores. Cells showed growth at 15-30 °C, pH 3.
View Article and Find Full Text PDFCRISPR/Cas13a-Programmed Cu NCs and -Scheme T-COF/AgS for Photoelectrochemical Biosensing of circRNA.
ACS Sens
January 2025
College of Public Health, Zhengzhou University, Zhengzhou 450001, China.
Circular RNAs (circRNAs), as a class of noncoding RNA molecules with a circular structure exhibit high stability and spatiotemporal-specific expression, making them ideal cancer biomarkers for liquid biopsy. Herein, a new photoelectrochemical (PEC) biosensor for a highly sensitive circRNA assay in the whole blood of lung cancer patients was designed based on CRISPR/Cas13a-programmed Cu nanoclusters (Cu NCs) and a -scheme covalent organic framework/silver sulfide (T-COF/AgS) composite. This -scheme T-COF/AgS composite accelerates electron transfer and produces an excellent initial photocurrent.
View Article and Find Full Text PDFIdentifying falsified COVID-19 vaccines by analysing vaccine vial label and excipient profiles using MALDI-ToF mass spectrometry.
NPJ Vaccines
January 2025
Department of Biochemistry, University of Oxford, OX1 3QU, Oxford, UK.
The rapid development and worldwide distribution of COVID-19 vaccines is a remarkable achievement of biomedical research and logistical implementation. However, these developments are associated with the risk of a surge of substandard and falsified (SF) vaccines, as illustrated by the 184 incidents with SF and diverted COVID-19 vaccines which have been reported during the pandemic in 48 countries, with a paucity of methods for their detection in supply chains. In this context, matrix-assisted laser desorption ionisation-time of flight (MALDI-ToF) mass spectrometry (MS) is globally available for fast and accurate analysis of bacteria in patient samples, offering a potentially accessible solution to identify SF vaccines.
View Article and Find Full Text PDFcircLOC375190 promotes autophagy through modulation of the mTORC1/TFEB axis in acute ischemic stroke-induced neurological injury.
Clinics (Sao Paulo)
January 2025
Department of Neurology, Daqing Oilfield General Hospital, Daqing City, Heilongjiang Province, China. Electronic address:
Objective: The authors explored differentially expressed circRNAs in Acute Ischemic Stroke (AIS) and revealed the role and potential downstream molecular mechanisms of circLOC375190.
Methods: circLOC375190 expression was modulated by lentiviral injection in the brain of transient Middle Cerebral Artery Occlusion (tMCAO) mice. Neurological dysfunction was assessed, as well as infarction size, histopathological changes, and neuronal apoptosis in tMCAO mice.
Want AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!