fluorescent dapi solution  (Thermo Fisher)

Journal: bioRxiv

Article Title: Nonlinear hippocampal coding of the pair-bonded partner in prairie voles

doi: 10.1101/2025.10.10.681589

Figure Lengend Snippet: a , Partner preference test. Top : Representative heatmaps depicting the proportion of time spent by male prairie voles at each location. Hot color indicates A partner female is located in a small chamber at one side, whereas a stranger female is in a chamber at the other side. The locations of the female voles are swapped from the Session 1 to 2 so that the possibility of place preference can be excluded. Bottom : The interaction time with the partner or stranger female, averaged across animals. Regardless of the female location, males exhibit a consistent preference for their partner ( n = 10 prairie voles, *** P < 0.001, paired t -test). b , Top : ( Left ) Schematic of retrograde tracer (CTB) injection into the NAc and representative image showing labeled neurons in vHPC ( red ). ( Right ) Schematic of anterograde tracer (AAV-hSyn-EGFP) injection into vHPC and resulting projections to the NAc ( green ). Bottom : Quantification of retrogradely labeled cells in the NAc- upstream regions ( n = 7 prairie voles). c , Muscimol or aCSF was bilaterally injected into the ventral hippocampus of male prairie voles. Top: ( Left ) Illustration of the injection experiment. ( Right ) Representative DAPI-stained brain slice showing the injection site in the ventral hippocampus; the dashed white line indicates cannula placement. Bottom : Experimental timeline of the pharmacological manipulation. d , Left : Preference index of male voles in each group. The preference index is defined as (partner interaction time – stranger interaction time) / (partner interaction time + stranger interaction time). The muscimol group shows a significantly reduced preference index compared to baseline, aCSF, and recovery groups ( n = 5 prairie voles, * P < 0.05, ** P < 0.01, paired t -test followed by Holm correction). Right : Locomotor activity of each group. No significant difference in locomotion across groups ( n = 5 prairie voles, paired t -test followed by Holm correction). Abbreviations : STR, stranger; PAR, partner; CTB, cholera toxin subunit B; NAc, nucleus accumbens; vHPC, ventral hippocampus; aCSF, artificial cerebrospinal fluid.

Article Snippet: Sections were stored in PBS, incubated in fluorescent DAPI solution (1:1000 in PBS; D1306, ThermoFisher) at room temperature for 90 min, washed in PBS for 10 min, and coverslipped using CC/Mount (Diagnostic BioSystems, USA).

Techniques: Injection, Labeling, Staining, Slice Preparation, Activity Assay

Journal: Theranostics

Article Title: Panax notoginseng -Derived Carbon Dots Herbzymes Ameliorate Renal Ischemia-Reperfusion Injury via Anti-Inflammation, Antioxidation and Gut-Kidney Axis Regulation

doi: 10.7150/thno.126643

Figure Lengend Snippet: Characterization of PN-CDs. (A) Images of TEM and HRTEM, and size distribution histogram of PN-CDs. (B) AFM and distribution height of PN-CDs. (C) UV spectra of PN-CDs. (D) The fluorescence spectra of PN-CDs. (E) Raman spectrum of PN-CDs. (F) XRD pattern of PN-CDs. (G) FT-IR spectrum of PN -CDs. (H) ¹H-NMR spectrum of PN-CDs. (I) XPS survey spectrum of PN-CDs. High-resolution XPS spectra of C 1s (J), O 1s (K), and N 1s (L) for PN-CDs. TEM: Transmission electron microscopy; HRTEM: High-resolution transmission electron microscopy; AFM: Atomic force microscopy. XRD: X-ray diffraction. FT-IR: Fourier-transform infrared spectroscopy. XPS: X-ray photoelectron spectroscopy.

Article Snippet: The ABTS assay kit, Anti fluorescence quenching sealing solution (with DAPI), Mito-Tracker Green, the JC-1 detection kit, along with the RNAeasyTM Animal RNA Isolation Kit with Spin Column, and dihydroethidium (DHE) probe were purchased from Beyotime (Shanghai, China).

Techniques: Fluorescence, Transmission Assay, Electron Microscopy, Microscopy, Fourier Transform Infrared Spectroscopy, Spectroscopy

Journal: Theranostics

Article Title: Panax notoginseng -Derived Carbon Dots Herbzymes Ameliorate Renal Ischemia-Reperfusion Injury via Anti-Inflammation, Antioxidation and Gut-Kidney Axis Regulation

doi: 10.7150/thno.126643

Figure Lengend Snippet: Cellular uptake and mitochondrial colocalization of PN-CDs. (A) Fluorescence images showing the time-dependent cellular uptake of PN-CDs in LPS-induced RAW 264.7 cells. Cells were stained with DAPI (blue, nuclei) and Actin (green, cytoplasm); PN-CDs were labeled with Cy5.5 (red). (B) Flow cytometry quantification of PN-CDs-Cy5.5 uptake efficiency in LPS-induced RAW 264.7 cells at different time points (0, 2, 4, 6, 8 h). (C) Fluorescence colocalization of PN-CDs (red) with mitochondria (green, labeled by Mito-Tracker Green) in RAW264.7 cells. Nuclei were stained with DAPI (blue). (D) Pearson's correlation coefficient analysis of PN-CDs and mitochondrial colocalization using Image J software. (E) Plot profile analysis of PN-CDs-Cy5.5 (red) and Mito-Tracker Green (green) fluorescence. (F) Fluorescence images showing the time-dependent cellular uptake of PN-CDs in HK-2 cells. (G) Flow cytometry quantification of PN-CDs-Cy5.5 uptake efficiency in HK-2 cells at different time points, with percentages of positive cells indicated. (H) Fluorescence colocalization of PN-CDs (red) with mitochondria (green) in HK-2 cells. (I) Pearson's correlation coefficient analysis of PN-CDs and mitochondrial colocalization in HK-2 cells. (J) Plot profile analysis of PN-CDs-Cy5.5 and Mito-Tracker Green fluorescence.

Article Snippet: The ABTS assay kit, Anti fluorescence quenching sealing solution (with DAPI), Mito-Tracker Green, the JC-1 detection kit, along with the RNAeasyTM Animal RNA Isolation Kit with Spin Column, and dihydroethidium (DHE) probe were purchased from Beyotime (Shanghai, China).

Techniques: Fluorescence, Staining, Labeling, Flow Cytometry, Software

Journal: Theranostics

Article Title: Panax notoginseng -Derived Carbon Dots Herbzymes Ameliorate Renal Ischemia-Reperfusion Injury via Anti-Inflammation, Antioxidation and Gut-Kidney Axis Regulation

doi: 10.7150/thno.126643

Figure Lengend Snippet: Anti-inflammatory and antioxidative stress activity of PN-CDs at the cellular level. (A) Fluorescence images showing intracellular ROS levels (green, DCFH-DA) and nuclear staining (blue, Hoechst) in RAW 264.7 cells treated with control, LPS, LPS+ PN-CDs1 (5 μg/mL), LPS + PN-CDs2 (10 μg/mL), or LPS+ N-acetylcysteine. (B) Flow cytometry analysis of intracellular ROS levels in RAW 264.7 cells under the above treatments. qPCR analysis of pro-inflammatory cytokine mRNA expression in LPS-induced RAW 264.7 cells. (C) TNF-α, (D) IL-1β, (E) IL-6. I: Control; II: LPS; III: LPS + PN-CDs1; IV: LPS + PN-CDs2, Ⅴ: LPS+ N-acetylcysteine. Data are presented as mean ± SD. Statistical significance: *** p < 0.001 vs. LPS group; ns: not significant. (F) Confocal fluorescence images showing intracellular ROS levels (green, DCFH-DA) and nuclear staining (blue, Hoechst) in RAW 264.7 cells treated with control, H₂O₂, H₂O₂ + PN-CDs1, H₂O₂ + PN-CDs2, or H 2 O 2 + N-acetylcysteine. (G) Flow cytometry analysis of intracellular ROS levels (DCFH-DA fluorescence, FITC-H channel) in H₂O₂-induced RAW 264.7 cells. (H) Calcein/PI fluorescence staining showing live cells (green, calcein) and dead/apoptotic cells (red, PI) in Control, H/R, H/R + low-concentration PN-CDs (PN-CDs1), and H/R + high-concentration PN-CDs (PN-CDs2) groups. (I) JC-1 mitochondrial membrane potential (ΔΨm) assay: Red fluorescence indicates J-aggregates (high ΔΨm), green fluorescence indicates monomers (low ΔΨm), and Merge images overlay red/green signals. Annexin V-FITC/PI flow cytometry analysis of apoptosis: (J) Representative scatter plots showing viable (Q4), early apoptotic (Q3), late apoptotic (Q2), and necrotic (Q1) cells; (K) Quantitative analysis of total apoptotic cells (Q2 + Q3). (L) CCK-8 assay showing cell viability in each group. qPCR analysis of renal tubular injury markers: (M) Kidney injury molecule-1 (KIM-1) and (N) neutrophil gelatinase-associated lipocalin (NGAL) mRNA expression. Flow cytometry analysis of intracellular ROS levels (DCFH-DA fluorescence, FITC channel): (O) Representative histograms; (P) Quantitative analysis of ROS-positive cells. Biochemical analysis of oxidative stress markers: (Q) Malondialdehyde (MDA) levels and (R) The ratio of glutathione (GSH)/glutathione (GSSG). (S) qPCR analysis of IL-6 mRNA expression in H/R model. Data are presented as mean ± SD. Statistical significance: ** p < 0.01, *** p < 0.001 vs. H/R group; ns, not significant. I: Control; II: H/R; III: H/R + PN-CDs1; IV: H/R + PN-CDs2, Ⅴ: H/R+ N-acetylcysteine.

Article Snippet: The ABTS assay kit, Anti fluorescence quenching sealing solution (with DAPI), Mito-Tracker Green, the JC-1 detection kit, along with the RNAeasyTM Animal RNA Isolation Kit with Spin Column, and dihydroethidium (DHE) probe were purchased from Beyotime (Shanghai, China).

Techniques: Activity Assay, Fluorescence, Staining, Control, Flow Cytometry, Expressing, Concentration Assay, Membrane, CCK-8 Assay

Journal: Theranostics

Article Title: Panax notoginseng -Derived Carbon Dots Herbzymes Ameliorate Renal Ischemia-Reperfusion Injury via Anti-Inflammation, Antioxidation and Gut-Kidney Axis Regulation

doi: 10.7150/thno.126643

Figure Lengend Snippet: In vivo biodistribution and retention of PN-CDs-Cy5.5. (A). Ex vivo organ fluorescence imaging of PN-CDs-Cy5.5 in control and RIRI model mice at 0 h, 0.5 h, 1 h, 2 h, 4 h, 8 h, 12 h, and 24 h post-intraperitoneal injection. Insets show magnified views of major organs: heart (H), liver (Li), spleen (S), lung (Lu), and kidney (K). (B-G) Quantitative analysis of mean fluorescence intensity (MFI) of PN-CDs-Cy5.5 in whole-body (B) and major organs, including heart (C), liver (D), spleen (E), lung (F), and kidney (G), at different time points. Data are presented as mean ± SD (n = 3 per group). * p < 0.05, ** p < 0.01, *** p < 0.001 vs. Control group. (H-I) Time-course profiles of MFI for liver (H) and kidney (I) in Control and RIRI groups, highlighting the delayed clearance and prolonged retention of PN-CDs-Cy5.5 in RIRI model mice.

Article Snippet: The ABTS assay kit, Anti fluorescence quenching sealing solution (with DAPI), Mito-Tracker Green, the JC-1 detection kit, along with the RNAeasyTM Animal RNA Isolation Kit with Spin Column, and dihydroethidium (DHE) probe were purchased from Beyotime (Shanghai, China).

Techniques: In Vivo, Ex Vivo, Fluorescence, Imaging, Control, Injection