Journal: Acta biochimica et biophysica Sinica

Article Title: Nutrition deficiency promotes apoptosis of cartilage endplate stem cells in a caspase-independent manner partially through upregulating BNIP3.

doi: 10.1093/abbs/gmw111

Figure Lengend Snippet: Figure 3. Nutrition deficiency mediates apoptosis partially through upregulating BNIP3 expression CESCs transfected with scramble or BNIP3 siRNA (h) were cultured under normal or nutrition-deficient condition for 48 h. (A) Total protein of CESCs was extracted after treatment and BNIP3 protein was detected by west- ern blot analysis. GAPDH was used as the internal reference. (B) Cell apoptosis was measured by staining with Annexin V-FITC/PI and analyzed by flow cytome- try. Data were presented as the mean ± SD. *P < 0.05, **P < 0.01. ND, nutrition deficiency.

Article Snippet: Cell apoptosis assay Cell apoptosis was analyzed using Annexin V-FITC Apoptosis Detection Kit (Beyotime, Shanghai, China).

Techniques: Expressing, Transfection, Cell Culture, Staining

Journal: Acta biochimica et biophysica Sinica

Article Title: Nutrition deficiency promotes apoptosis of cartilage endplate stem cells in a caspase-independent manner partially through upregulating BNIP3.

doi: 10.1093/abbs/gmw111

Figure Lengend Snippet: Figure 2. Nutrition deficiency promotes apoptosis of CESCs and BNIP3 expression CESCs were cultured under normal or nutrition-deficient condition for 24 or 48 h. (A) Sketch of experimental design. (B) Representative images photographed under a light microscope (×100) demonstrated the effects of nutrition defi- ciency on morphology of CESCs. Scale bar, 50 μm. (C) Cell apoptosis was measured by staining with Annexin V-FITC/PI and analyzed by flow cytometry. (D) BNIP3 mRNA quantification (normalized to GAPDH) in CESCs post nutrition deficiency. (E) Total protein of CESCs was extracted after treatment and BNIP3 pro- tein was detected by western blot analysis. GAPDH was used as the internal reference. Data were presented as the mean ± SD. *P < 0.05, **P < 0.01. ND, nutri- tion deficiency.

Article Snippet: Cell apoptosis assay Cell apoptosis was analyzed using Annexin V-FITC Apoptosis Detection Kit (Beyotime, Shanghai, China).

Techniques: Expressing, Cell Culture, Light Microscopy, Staining, Cytometry, Western Blot

Journal: Frontiers in Oncology

Article Title: Novel Breast-Specific Long Non-coding RNA LINC00993 Acts as a Tumor Suppressor in Triple-Negative Breast Cancer

doi: 10.3389/fonc.2019.01325

Figure Lengend Snippet: LINC00993 suppresses the growth of triple-negative breast cancer (TNBC) cells in vitro . (A) Structure of LINC00993 expression plasmid for adenovirus. (B) LINC00993 expression adenovirus infection efficiency showed by fluorescence microscope in MDA-MB-231 cells. Original magnification, ×400. Scale bars, 50 μm. (C) Expression of LINC00993 detected by qRT-PCR. MDA-MB-231 cells were infected by adenovirus for 24 h, and RNA was extracted. (D) Image of clone formation assay. (E) Number of clones were counted 2 weeks after plantation. (F) MDA-MB-231 cells were planted into 24-well plates. Twenty-four hours later, adenovirus was added to each well. Three wells of cells were digested and counted every 24 h. (G) LINC00993 expression caused apoptosis shown by TUNEL assay. Green points reflected apoptosis, and we used DAPI to stain DNA. Positive control cells were treated with DNase I, negative control cells were collected without adding TUNEL reaction buffer. Original magnification, ×100. Scale bars, 50 μm. (H) Effect of LINC00993 on cell cycle detected by flow cytometry. (I) Flow cytometry cell cycle results shown in a bar plot. (J) Invasive ability tested by Transwell assay. Twenty thousand cells were plated in each well, cells were observed after 24 h of incubation. (K) Bar plot for the number of cells that migrated across the membrane. *** P < 0.001, based on Student's t -test. Data were presented as mean ± SEM.

Article Snippet: To analyze cell apoptosis, terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) assays were performed with One Step TUNEL Apoptosis Assay Kit (FITC, Beyotime, China) according to the manufacturer's instructions.

Techniques: In Vitro, Expressing, Plasmid Preparation, Infection, Fluorescence, Microscopy, Quantitative RT-PCR, Tube Formation Assay, Clone Assay, TUNEL Assay, Staining, Positive Control, Negative Control, Flow Cytometry, Transwell Assay, Incubation, Membrane

Journal: Physiologia plantarum

Article Title: The interplay of hydraulic failure and cell vitality explains tree capacity to recover from drought.

doi: 10.1111/ppl.13331

Figure Lengend Snippet: FIGURE 5 Cross-sections of Prunus lusitanica (A and B) and Pseudotsuga menziesii (C and D) stems in control conditions. Cross-sections were stained using fluorescein diacetate (FDA) (60 μm thick cross-section—1% solution) and microphotographs were taken using a bright light (A and C) and an inverted fluorescence microscope (YFP filter; B and D). Fluorescent spots indicate living cells

Article Snippet: Cross-sections were observed using an inverted fluorescence microscope (Axio Observer Z1, ZEISS; Bright light or YFP filter) within the next hour after staining.

Techniques: Control, Staining, Fluorescence, Microscopy

Journal: Biomaterials science

Article Title: A Chimeric, Multivalent Assembly of Galectin-1 and Galectin-3 with Enhanced Extracellular Activity

doi: 10.1039/c8bm01631c

Figure Lengend Snippet: Phosphatidylserine exposure determined via Annexin V-FITC staining (green) and membrane permeability determined via propidium iodide (PI) (red) after Jurkat T cells were treated with (A) 5 μM or (B) 0.5 μM G1, G1-PEG-G1, G1/G3, or G1/G3 Zipper for 4 h. Brightfield images in top row are representative of extent of agglutination before treatment with β-lactose. * in the micrographs denotes cell agglutination Single channel fluorescence images are available in Supplementary Information Fig. 5. Scale bars = 50 μm.

Article Snippet: Images were taken on a Zeiss Axio Observer inverted microscope with FITC (excitation = 470/40 nm and emission = 525/50 nm) and rhodamine (excitation = 546/12 nm and emission = 575–640 nm) filter sets.

Techniques: Staining, Membrane, Permeability, Agglutination, Fluorescence

Journal: Antioxidants

Article Title: A Multi-Target Pharmacological Correction of a Lipoyltransferase LIPT1 Gene Mutation in Patient-Derived Cellular Models

doi: 10.3390/antiox13081023

Figure Lengend Snippet: The analysis of iron accumulation in mutant LIPT1 fibroblasts. ( A ). The control and patient’s fibroblasts (LIPT1) were stained with Prussian Blue staining. Mutant cells were treated with 100 µM Deferiprone (DEF). Images were acquired by a Zeiss Axio Vert A1 microscope. Scale bar: 20 µm. ( B ). The quantification of Prussian Blue staining-integrated density. Images were analyzed by the ImageJ software (at least 30 images were analyzed per each condition and experiment). ( C ). The quantification of iron content by ICP-MS. ( D ). Lipofuscin accumulation was assessed by Sudan Black staining. Mutant cells were treated with 100 µM DEF. Images were acquired by a Zeiss Axio Vert A1 microscope. A PKAN (pantothenate kinase-associated neurodegeneration) cell line was used as a positive control of lipofuscin accumulation. Scale bar: 20 µm. ( E ). The quantification of Sudan Black staining-integrated density (at least 30 images were analyzed per each condition and experiment). Data represent the mean ± SD of 3 independent experiments. **** p < 0.0001 between the control and patient’s fibroblasts. #### p < 0.0001 between mutant fibroblasts untreated and treated with Deferiprone. a.u.: arbitrary units.

Article Snippet: Lipid peroxidation in fibroblasts was evaluated using a light and fluorescence Axio Vert A1 microscope (Zeiss, Oberkochen, Germany) with a 20x objective.

Techniques: Mutagenesis, Control, Staining, Microscopy, Software, Positive Control

Journal: Antioxidants

Article Title: A Multi-Target Pharmacological Correction of a Lipoyltransferase LIPT1 Gene Mutation in Patient-Derived Cellular Models

doi: 10.3390/antiox13081023

Figure Lengend Snippet: The effect of CocT on iron accumulation in mutant LIPT1 fibroblasts. ( A ). Untreated and treated (for seven days) control and patient (LIPT1) fibroblasts were stained with Prussian Blue staining. ( B ). Lipofuscin accumulation was assessed by Sudan Black staining. A PKAN cell line was used as a positive control of lipofuscin accumulation . Mutant cells were treated with 100 µM DEF. Representative images were acquired by a Zeiss Axio Vert A1 microscope (at least 30 images were analyzed per each condition and experiment). Scale bar: 20 µm. ( C ). The quantification of Prussian Blue staining-integrated density. Images were analyzed by ImageJ software (at least 30 images were analyzed per each condition and experiment). ( D ). The quantification of iron content by ICP-MS. ( E ). The quantification of Sudan Black staining-integrated density. Data represent the mean ± SD of 3 independent experiments. **** p < 0.0001 between control and mutant LIPT1 fibroblasts. #### p < 0.0001 between untreated and treated mutant LIPT1 fibroblasts. aaaa p < 0.0001 between untreated and treated mutant LIPT1 fibroblasts with Deferiprone. a.u.: arbitrary units.

Article Snippet: Lipid peroxidation in fibroblasts was evaluated using a light and fluorescence Axio Vert A1 microscope (Zeiss, Oberkochen, Germany) with a 20x objective.

Techniques: Mutagenesis, Control, Staining, Positive Control, Microscopy, Software

Journal: Antioxidants

Article Title: A Multi-Target Pharmacological Correction of a Lipoyltransferase LIPT1 Gene Mutation in Patient-Derived Cellular Models

doi: 10.3390/antiox13081023

Figure Lengend Snippet: The effect of CocT on iron accumulation in iNs generated from control and patient-derived fibroblasts by direct reprogramming. Control and mutant (LIPT1) iNs were treated with CocT for seven days. ( A ). Representative images were acquired by a Zeiss Axio Vert A1 microscope. Tau was used as a neuronal marker. Scale bar: 15 μm. ( B ). Quantification of Prussian Blue staining images were obtained by Image J software (at least 30 images were analyzed per each experimental condition). **** p < 0.0001 between control and mutant LIPT1 iNs. #### p < 0.0001 between untreated and treated mutant LIPT1 iNs. a.u.: arbitrary units.

Article Snippet: Lipid peroxidation in fibroblasts was evaluated using a light and fluorescence Axio Vert A1 microscope (Zeiss, Oberkochen, Germany) with a 20x objective.

Techniques: Generated, Control, Derivative Assay, Mutagenesis, Microscopy, Marker, Staining, Software

Journal: Scientific Reports

Article Title: Application of corneal injury models in dual fluorescent reporter transgenic mice to understand the roles of the cornea and limbus in angiogenic and lymphangiogenic privilege

doi: 10.1038/s41598-019-48811-z

Figure Lengend Snippet: Axio Zoom images of mouse eyes as well as quantitated areas of corneal HA and LA after debridement of a 1.5-mm-diameter circle in the center of the cornea. Visible but minimal neovascularization, particularly HA, was observed. ([ A ] comparison of dorsal HA on day 0 and day 14 (p = 0.0391), comparison of dorsal LA on day 0 and day 14 (p = 0.3831); [ B ] comparison of ventral HA on day 0 and day 14 (p = 0.1887), comparison of ventral LA on day 0 and day 14 (p = 0.4765). n = 5 ).

Article Snippet: The injured eyes were visualized under an Axio Zoom V16 fluorescence microscope (Zeiss Microscopy, Oberkochen, Germany) to assess corneal HA and LA at baseline (before injury) and days 4, 7, 10, and 14 after injury.

Techniques: Comparison

Journal: Scientific Reports

Article Title: Application of corneal injury models in dual fluorescent reporter transgenic mice to understand the roles of the cornea and limbus in angiogenic and lymphangiogenic privilege

doi: 10.1038/s41598-019-48811-z

Figure Lengend Snippet: Axio Zoom images of mouse eyes as well as quantitated areas of corneal HA and LA after whole corneal epithelium debridement. Enhanced corneal HA and LA through day 14 were observed from both the dorsal and ventral views. ([ A ] comparison of dorsal HA on day 0 and day 14 (p = 0.0072), comparison of dorsal LA on day 0 and day 14 (p = 0.1698); [ B ] comparison of ventral HA on day 0 and day 14 (p = 0.0198), comparison of ventral LA on day 0 and day 14 (p = 0.2330). n = 3).

Article Snippet: The injured eyes were visualized under an Axio Zoom V16 fluorescence microscope (Zeiss Microscopy, Oberkochen, Germany) to assess corneal HA and LA at baseline (before injury) and days 4, 7, 10, and 14 after injury.

Techniques: Comparison

Journal: Scientific Reports

Article Title: Application of corneal injury models in dual fluorescent reporter transgenic mice to understand the roles of the cornea and limbus in angiogenic and lymphangiogenic privilege

doi: 10.1038/s41598-019-48811-z

Figure Lengend Snippet: Axio Zoom images of mouse eyes as well as quantitated areas of corneal HA and LA after superficial debridement of the whole limbal plus whole corneal. Enhanced corneal HA and LA through day 14 were observed from both the dorsal and ventral views. ([ A ] comparison of dorsal HA on day 0 and day 14 (p = 0.0014), comparison of dorsal LA on day 0 and day 14 (p = 0.3779); [ B ] comparison of ventral HA on day 0 and day 14 (p = 0.0080), comparison of ventral LA on day 0 and day 14 (p = 0.9251). n = 6).

Article Snippet: The injured eyes were visualized under an Axio Zoom V16 fluorescence microscope (Zeiss Microscopy, Oberkochen, Germany) to assess corneal HA and LA at baseline (before injury) and days 4, 7, 10, and 14 after injury.

Techniques: Comparison

Journal: Scientific Reports

Article Title: Application of corneal injury models in dual fluorescent reporter transgenic mice to understand the roles of the cornea and limbus in angiogenic and lymphangiogenic privilege

doi: 10.1038/s41598-019-48811-z

Figure Lengend Snippet: Axio Zoom images of mouse eyes in which the whole limbal epithelium was debrided and quantitated areas of corneal HA and LA. In this model, no discernible corneal HA or LA was observed. ([ A ] comparison of dorsal HA on day 0 and day 14 (p = 0.2412), comparison of dorsal LA on day 0 and day 14 (p = 0.5079); [ B ] comparison of ventral HA on day 0 and day 14 (p = 0.1682), comparison of ventral LA on day 0 and day 14 (p = 0.2740). n = 4).

Article Snippet: The injured eyes were visualized under an Axio Zoom V16 fluorescence microscope (Zeiss Microscopy, Oberkochen, Germany) to assess corneal HA and LA at baseline (before injury) and days 4, 7, 10, and 14 after injury.

Techniques: Comparison

Journal: Scientific Reports

Article Title: Application of corneal injury models in dual fluorescent reporter transgenic mice to understand the roles of the cornea and limbus in angiogenic and lymphangiogenic privilege

doi: 10.1038/s41598-019-48811-z

Figure Lengend Snippet: Axio Zoom images of mouse eyes as well as quantitated areas of corneal HA and LA after deep debridement of the dorsal half of the limbal plus whole corneal epithelium. From the dorsal view, both blood and lymphatic vessels regrew toward but not beyond the baseline. The ventral view exhibited growth of both blood and lymphatic vessels surpassing the baseline from day 7 and onward. ([ A ] comparison of dorsal HA on day 0 and day 14 (p = 0.6765), comparison of dorsal LA on day 0 and day 14 (p = 0.1824); [ B ] comparison of ventral HA on day 0 and day 14 (p = 0.0054), comparison of ventral LA on day 0 and day 14 (p = 0.4274). n = 5).

Article Snippet: The injured eyes were visualized under an Axio Zoom V16 fluorescence microscope (Zeiss Microscopy, Oberkochen, Germany) to assess corneal HA and LA at baseline (before injury) and days 4, 7, 10, and 14 after injury.

Techniques: Comparison

Journal: Scientific Reports

Article Title: Application of corneal injury models in dual fluorescent reporter transgenic mice to understand the roles of the cornea and limbus in angiogenic and lymphangiogenic privilege

doi: 10.1038/s41598-019-48811-z

Figure Lengend Snippet: Axio Zoom images of mouse eyes as well as quantitated areas of corneal HA and LA after whole corneal debridement in CDh5-CreERT2VEGFR2lox/PGFD mice. Significant HA and LA were observed in the control group from Day 4 and Day 10, respectively ([ A ] dorsal view, n = 4). Meanwhile, corneal HA and LA in response to whole corneal epithelium debridement were abolished on days 4, 7, 10, and 14 in tamoxifen-treated CDh5-CreERT2 VEGFR2lox/PGFD mice ([ B ] dorsal view, n = 3).

Article Snippet: The injured eyes were visualized under an Axio Zoom V16 fluorescence microscope (Zeiss Microscopy, Oberkochen, Germany) to assess corneal HA and LA at baseline (before injury) and days 4, 7, 10, and 14 after injury.

Techniques: Control

Journal: Scientific reports

Article Title: Optogenetics design of mechanistically-based stimulation patterns for cardiac defibrillation.

doi: 10.1038/srep35628

Figure Lengend Snippet: Figure 1. Simultaneous all-optical map and control of cardiac conduction pathway. (a) Scheme of the wide-field fluorescence macroscope consisting of a LED at a wavelength centered at 625 nm for excitation of the voltage-sensitive dye and a CW laser at 473 nm for ChR2 activation. The system is provided with a random access scanning head comprised of two orthogonally mounted acousto-optical deflectors (AOD x-y). A 1.25x objective is used for imaging and stimulation of the whole mouse heart, a 20x objective is used to focus the fluorescence signal into a central portion (100 × 100 pixel) of a sCMOS sensor operating at a frame rate of 2 kHz. (b) Electrocardiographic recording of a ChR2-expressing heart, showing sinus rhythm with a frequency of 3 Hz. Blue arrowheads represent the ChR2 stimulation performed at 5 Hz (laser power at the sample 4.9 mW, illumination time 5 ms). The orange line represents the LED illumination. (c) Fluorescence image (F0) of a ChR2-expressing heart stained with voltage sensitive dye (left). Scale bar of 2 mm in yellow. Six representative frames of optical mapping (ΔF/F0) recorded from the same heart. The electrical activation is reported in red and the baseline in cyan. On the right, corresponding color-scaled isochronal map of the action potential reporting the voltage activation time per pixel. (d) On the left, fluorescence image (F0) of a ChR2-expressing heart stimulated in the apex either with an electrode (red spot, above) or with ChR2 activation with the blue laser (blue spot, below). The green frame represents the AODs working-field on the sample. Corresponding optical mapping frames (ΔF/F0) recorded from the same heart are reported next. On the right, corresponding color-scaled isochronal map of the action potential reporting the activation time per pixel.

Article Snippet: The whole mouse heart was excited in wide-field configuration using a 1.25x objective (EC Plan-Neofluar 1.25x/0.03 M27, Carl Zeiss Microscopy) and a LED operating at a wavelength centered at 625 nm (M625L3, Thorlabs) followed by a band-pass filter at 640/40 nm (FF01640/40-25, Semrock).

Techniques: Control, Fluorescence, Activation Assay, Imaging, Expressing, Staining