Current Protocols
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2026
·Vol. 6
·期 9
·e70458
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IF 2.5
Di Wang,Qiqi Lv,Shichao Lin
Single-cell RNA sequencing (scRNA-seq) reveals the transcriptional heterogeneity of cells, revolutionizing our understanding of cellular processes. However, the static snapshots obtained from scRNA-seq fail to reveal the time-resolved dynamics of transcription, which impedes critical insights into various biological processes, such as cellular differentiation, embryonic development, disease progression, and responses to external stimuli. Here, we describe Well-TEMP-seq, a protocol for massively parallel profiling of the temporal dynamics of single-cell gene expression. Well-TEMP-seq combines metabolic RNA labeling with a microwell-based scRNA-seq method, Well-paired-seq, to distinguish newly transcribed RNAs marked by T-to-C substitutions from pre-existing RNAs in each of thousands of single cells. Well-TEMP-seq is high-throughput, cost-effective, accurate, and provides a low cell loss rate and high single cell/bead pairing efficiency. More importantly, Well-TEMP-seq can be easily set up in other labs, and the loading of cells and beads can be easily accomplished by an optical microscope and a pipette. We believe that Well-TEMP-seq will be widely adopted and help researchers perform transformative research to unveil the dynamics of single-cell gene expression in diverse biological processes. © 2026 Wiley Periodicals LLC. Basic Protocol 1 : Well-paired-seq chip fabrication Basic Protocol 2 : Well-TEMP-seq sample processing Basic Protocol 3 : Bioinformatics analysis
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PubMed
高影响因子
RSC Advances
·
2020
·Vol. 10
·期 23
·13830-13837
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IF 3.119
Ruifen Peng,Yuxia Pang,Xueqing Qiu,Yong Qian,Mingsong Zhou
In the formulation of pesticide Suspension Concentrate (SC), some photosensitive pesticides are easily decomposed in the preparation. In this study, a hindered amine modified lignosulfonate (SL-Temp) with anti-photolysis function was synthesized using 4-amino-2,2,6,6-tetramethylpiperidine (Temp) and Sodium Lignosulfonate (SL) to solve this problem. The obtained SL-Temp was used as a dispersant to prepare 5% SC of avermectin, which shows good physical stability. The decomposition rate of the avermectin in SC after accelerating hot storage is 0%, which is much lower than 6.1% when SL was used as the dispersant. After being exposed to UV irradiation for 60 hours, the highest retention rate of avermectin is 87.1% when SL-Temp was used as the dispersant, which is much higher than 73.6% when SL was used as the dispersant, and also higher than 76.3% when a small molecule antioxidant (BHT) was added to the formulation. QCM-D studies revealed that the SL-Temp adsorption layer on avermectin particles can compete to absorb partial ultraviolet rays, hinder the penetration of ultraviolet light, and scavenge the free radicals produced by photooxidation, so as to protect avermectin from degradation.
研究领域:
农药科学
·
材料化学
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高影响因子
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
·
2023
·Vol. 120
·期 30
·e2305706120
·
IF 11.1
Jing-Hang Wu,Fei Chen,Tian-Hao Yang,Han-Qing Yu
Singlet oxygen (1O2) plays a pivotal role in numerous catalytic oxidation processes utilized in water purification and chemical synthesis. The spin-trapping method based on electron paramagnetic resonance (EPR) analysis is commonly employed for 1O2 detection. However, it is often limited to time-independent acquisition. Recent studies have raised questions about the reliability of the 1O2 trapper, 2,2,6,6-tetramethylpiperidine (TEMP), in various systems. In this study, we introduce a comprehensive, kinetic examination to monitor the spin-trapping process in EPR analysis. The EPR intensity of the trapping product was used as a quantitative measurement to evaluate the concentration of 1O2 in aqueous systems. This in situ kinetic study was successfully applied to a classical photocatalytic system with exceptional accuracy. Furthermore, we demonstrated the feasibility of our approach in more intricate 1O2-driven catalytic oxidation processes for water decontamination and elucidated the molecular mechanism of direct TEMP oxidation. This method can avoid the false-positive results associated with the conventional 2D 1O2 detection techniques, and provide insights into the reaction mechanisms in 1O2-dominated catalytic oxidation processes. This work underscores the necessity of kinetic studies for spin-trapping EPR analysis, presenting an avenue for a comprehensive exploration of the mechanisms governing catalytic oxidation processes.
研究领域:
分析化学
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环境科学
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催化
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物理化学
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DOI
PubMed
高影响因子
EUROPEAN POLYMER JOURNAL
·
2023
·
IF 6
Jingjing Wang,Zhaoqi Pan,Jie Liu,Qun Shao,Yuan Liang,Shuohan Huang,Wenbin Jin,Zefeng Li,Zhenfei Zhang,Chunlin Ye,Yuwei Chen,Peng Wei,Yanping Wang,Yong He,Yumin Xia
In this study, a polymeric ionic liquid (PIL) called Poly[ECH-BIM]Cl was polymerized by ring-opening and quaternization reaction of benzimidazole and epichlorohydrin without additional ion-exchange reaction. Both 1 H NMR and fourier transform infrared (FTIR) confirmed the chemical structure of Poly[ECH-BIM]Cl. Poly[ECH-BIM]Cl in water was thermoresponsive and its UCST transition was studied by UV–vis spectrometry, dynamic light scattering (DLS) and scanning electron microscopy (SEM) at variable temperature. Moreover, the first introduced Temp-resistance behavior during ions pairing found Poly[ECH-BIM]Cl had properties similar to those of the precipitation of inorganic salts from their saturated solutions. The variable-temperature 1 H NMR analysis demonstrated that the tailored interchain π-π interactions promoted the aggregation of macromolecular chains and particles formed from Poly[ECH-BIM]Cl aqueous solution. The results showed that increasing the interaction between cations of PIL can endow its thermosensibility and the possibility of being a programmable shuttle.
研究领域:
材料科学
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化学
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高影响因子
Journal of Cleaner Production
·
2023
·
IF 11.1
Shicheng Yuan,Hui Hu,Hao Huang,Zhongming Chen,Jiaxin Chen,Mi Zhang,Kunpeng Li,Ting Zhou,Ruibin Lv
Chlorobenzene (CB) is an important organic chemical material that is volatile and difficult to degrade, and has received much attention because of its strong ozone generation potential. An electrochemical-sodium peroxydisulfate (PDS) system was constructed and the oxidation reaction path of chlorobenzene in this system was analyzed. The effects of current density, PDS concentration, initial pH, CB blow-off gas ratio and temperature on the oxidation and mineralization rate of chlorobenzene in the system were investigated respectively. At a current density of 168 A/m 2 , a PDS concentration of 0.5 mol/L, an initial pH of 10, a CB blow-off gas ratio of 20% and a reaction temperature of 60 °C, the CB conversion and mineralization rates were able to reach more than 90%. The results of electron paramagnetic resonance (EPR) experiments using 5,5-dimethyl-1-pyrroline N-oxide (DMPO) and 2,2,6,6-tetramethyl-4-piperidone hydrochloride (TEMP) as spin trapping agents indicate the presence of ⦁OH, · S O 4 − and 1 O 2 in the system. The signal of · O 2 − was also captured after the quenching of ⦁OH, which further indicates the presence of 1 O 2 . And the results of active substance quenching experiments showed that the active species that mainly contributed to the oxidation of chlorobenzene included ⦁OH, · S O 4 − and 1 O 2 , and their contributions to the oxidation of chlorobenzene were 25.0%, 31.7% and 22.2%, respectively. The Liquid Chromatograph Mass Spectrometer (LC-MS) characterization results showed that the formation of chlorophenol and hydroquinone and their ring-opening reactions were the most critical processes in the oxidative decomposition of chlorobenzene, and the reaction pathways for the oxidation of chlorobenzene by ⦁OH, · S O 4 − and 1 O 2 were proposed by these characterization results.
研究领域:
环境科学
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化工工程
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分析化学
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高影响因子
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