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hypertonic lysis buffer per plate  (ArcticZymes)


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    ArcticZymes hypertonic lysis buffer per plate
    Hypertonic Lysis Buffer Per Plate, supplied by ArcticZymes, used in various techniques. Bioz Stars score: 94/100, based on 78 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/hypertonic+buffer/SAN+HQ/pm40744920-246-26-41
    Average 94 stars, based on 78 article reviews
    hypertonic lysis buffer per plate - by Bioz Stars, 2026-09
    94/100 stars

    Images

    Related Articles

    Blocking Assay:

    Article Title: Regionally mapped astrocytic responses to cortical and white matter stroke show differential roles in astrocyte-induced vascular remodeling.
    Article Snippet: Article Regionally mapped astrocytic responses to cortical and white matter stroke show differential roles in astrocyte-induced vascular remodeling

    Hybridization:

    Article Title: Regionally mapped astrocytic responses to cortical and white matter stroke show differential roles in astrocyte-induced vascular remodeling.
    Article Snippet: Article Regionally mapped astrocytic responses to cortical and white matter stroke show differential roles in astrocyte-induced vascular remodeling

    Amplification:

    Article Title: Regionally mapped astrocytic responses to cortical and white matter stroke show differential roles in astrocyte-induced vascular remodeling.
    Article Snippet: Article Regionally mapped astrocytic responses to cortical and white matter stroke show differential roles in astrocyte-induced vascular remodeling

    Electron Microscopy:

    Article Title: Regionally mapped astrocytic responses to cortical and white matter stroke show differential roles in astrocyte-induced vascular remodeling.
    Article Snippet: Article Regionally mapped astrocytic responses to cortical and white matter stroke show differential roles in astrocyte-induced vascular remodeling

    Antibody Labeling:

    Article Title: Regionally mapped astrocytic responses to cortical and white matter stroke show differential roles in astrocyte-induced vascular remodeling.
    Article Snippet: Article Regionally mapped astrocytic responses to cortical and white matter stroke show differential roles in astrocyte-induced vascular remodeling

    Immunoprecipitation:

    Article Title: Regionally mapped astrocytic responses to cortical and white matter stroke show differential roles in astrocyte-induced vascular remodeling.
    Article Snippet: Article Regionally mapped astrocytic responses to cortical and white matter stroke show differential roles in astrocyte-induced vascular remodeling

    RNA Sequencing:

    Article Title: Regionally mapped astrocytic responses to cortical and white matter stroke show differential roles in astrocyte-induced vascular remodeling.
    Article Snippet: Article Regionally mapped astrocytic responses to cortical and white matter stroke show differential roles in astrocyte-induced vascular remodeling

    In Vivo:

    Article Title: Efficient in vivo generation of CAR T cells using a retargeted fourth-generation lentiviral vector.
    Article Snippet: .. Large-scale preparations (5 L) used for the in vivo study were performed in the Applikon EZ stirred-tank bioreactor, treated with Benzonase (Merck Life Science, Gillingham, UK), and clarified and purified using anion exchange chromatography on an AKTA Pure (Cytiva, Amersham, UK) using the Mustang Q XT5 membrane (Cytiva, Amersham, UK); eluted fractions were treated with Salt Activated Nuclease (ArcticZymes Technologies ASA, Tromso, Norway), diluted to 300 mM NaCl, and concentrated 1,000-fold by centrifugation as detailed above. .. Vector pellets were finally resuspended in TSSM buffer (sodium chloride [100 mM], Tris, pH 7.3 [20 mM], sucrose [10 mg/mL], and mannitol [10 mg/mL]), aliquoted, and stored at − 80◦C.

    Purification:

    Article Title: Efficient in vivo generation of CAR T cells using a retargeted fourth-generation lentiviral vector.
    Article Snippet: .. Large-scale preparations (5 L) used for the in vivo study were performed in the Applikon EZ stirred-tank bioreactor, treated with Benzonase (Merck Life Science, Gillingham, UK), and clarified and purified using anion exchange chromatography on an AKTA Pure (Cytiva, Amersham, UK) using the Mustang Q XT5 membrane (Cytiva, Amersham, UK); eluted fractions were treated with Salt Activated Nuclease (ArcticZymes Technologies ASA, Tromso, Norway), diluted to 300 mM NaCl, and concentrated 1,000-fold by centrifugation as detailed above. .. Vector pellets were finally resuspended in TSSM buffer (sodium chloride [100 mM], Tris, pH 7.3 [20 mM], sucrose [10 mg/mL], and mannitol [10 mg/mL]), aliquoted, and stored at − 80◦C.

    Chromatography:

    Article Title: Efficient in vivo generation of CAR T cells using a retargeted fourth-generation lentiviral vector.
    Article Snippet: .. Large-scale preparations (5 L) used for the in vivo study were performed in the Applikon EZ stirred-tank bioreactor, treated with Benzonase (Merck Life Science, Gillingham, UK), and clarified and purified using anion exchange chromatography on an AKTA Pure (Cytiva, Amersham, UK) using the Mustang Q XT5 membrane (Cytiva, Amersham, UK); eluted fractions were treated with Salt Activated Nuclease (ArcticZymes Technologies ASA, Tromso, Norway), diluted to 300 mM NaCl, and concentrated 1,000-fold by centrifugation as detailed above. .. Vector pellets were finally resuspended in TSSM buffer (sodium chloride [100 mM], Tris, pH 7.3 [20 mM], sucrose [10 mg/mL], and mannitol [10 mg/mL]), aliquoted, and stored at − 80◦C.

    Membrane:

    Article Title: Efficient in vivo generation of CAR T cells using a retargeted fourth-generation lentiviral vector.
    Article Snippet: .. Large-scale preparations (5 L) used for the in vivo study were performed in the Applikon EZ stirred-tank bioreactor, treated with Benzonase (Merck Life Science, Gillingham, UK), and clarified and purified using anion exchange chromatography on an AKTA Pure (Cytiva, Amersham, UK) using the Mustang Q XT5 membrane (Cytiva, Amersham, UK); eluted fractions were treated with Salt Activated Nuclease (ArcticZymes Technologies ASA, Tromso, Norway), diluted to 300 mM NaCl, and concentrated 1,000-fold by centrifugation as detailed above. .. Vector pellets were finally resuspended in TSSM buffer (sodium chloride [100 mM], Tris, pH 7.3 [20 mM], sucrose [10 mg/mL], and mannitol [10 mg/mL]), aliquoted, and stored at − 80◦C.

    Centrifugation:

    Article Title: Efficient in vivo generation of CAR T cells using a retargeted fourth-generation lentiviral vector.
    Article Snippet: .. Large-scale preparations (5 L) used for the in vivo study were performed in the Applikon EZ stirred-tank bioreactor, treated with Benzonase (Merck Life Science, Gillingham, UK), and clarified and purified using anion exchange chromatography on an AKTA Pure (Cytiva, Amersham, UK) using the Mustang Q XT5 membrane (Cytiva, Amersham, UK); eluted fractions were treated with Salt Activated Nuclease (ArcticZymes Technologies ASA, Tromso, Norway), diluted to 300 mM NaCl, and concentrated 1,000-fold by centrifugation as detailed above. .. Vector pellets were finally resuspended in TSSM buffer (sodium chloride [100 mM], Tris, pH 7.3 [20 mM], sucrose [10 mg/mL], and mannitol [10 mg/mL]), aliquoted, and stored at − 80◦C.

    Article Title: GFP-on mouse model for interrogation of in vivo gene editing.
    Article Snippet: .. 4 days after transfection, cells and media were collected using a cell scraper (Corning, Glendale, Arizona), pelleted by centrifugation for 2000×g for 10min, resuspended in 500μL hypertonic lysis buffer per plate (40mM Tris base, 500mM NaCl, 2mM MgCl2, 100U/mL salt-active nuclease (ArcticZymes; Wayne, PA)) and incubated at 37 °C for 1 h to lyse the cells. .. The media was decanted and combined with a 5x solution of 40% poly(ethylene glycol) 8000 (PEG8k) (Sigma-Aldrich, St. Louis,MO) in 2.5MNaCl (final concentration 8% PEG, 500mM NaCl).

    Incubation:

    Article Title: A sensitive orange fluorescent calcium ion indicator for imaging neural activity
    Article Snippet: .. The lysate was incubated with 100 U/mL salt-active nuclease (Arcticzymes) at 37°C for 1 hour and then centrifuged at 2,000 g for 15 minutes. .. AAV was purified from the resulting lysate using an iodixanol step gradient containing 15%, 25%, 40%, and 60% iodixanol in OptiSeal tubes (Beckman Coulter) followed by centrifugation at 350,000 g using a Type 70 Ti ultracentrifuge rotor (Beckman Coulter).

    Article Title: GFP-on mouse model for interrogation of in vivo gene editing.
    Article Snippet: .. 4 days after transfection, cells and media were collected using a cell scraper (Corning, Glendale, Arizona), pelleted by centrifugation for 2000×g for 10min, resuspended in 500μL hypertonic lysis buffer per plate (40mM Tris base, 500mM NaCl, 2mM MgCl2, 100U/mL salt-active nuclease (ArcticZymes; Wayne, PA)) and incubated at 37 °C for 1 h to lyse the cells. .. The media was decanted and combined with a 5x solution of 40% poly(ethylene glycol) 8000 (PEG8k) (Sigma-Aldrich, St. Louis,MO) in 2.5MNaCl (final concentration 8% PEG, 500mM NaCl).

    Article Title: AAV yield, bioactivity, and particle heterogeneity are impacted by genome size and non-coding DNA elements
    Article Snippet: .. Genomic DNA and remaining plasmid DNA were digested by incubation of the lysate with salt-active nuclease (ArcticZymes; Tromso, Norway) for 1 h, followed by polyethylene glycol (PEG) precipitation of proteins, including AAVs, for 3 h. The PEG pellet was resuspended overnight, and AAVs were purified by an iodixanol gradient to remove contaminating proteins and empty capsids. .. After exchanging the buffer and simultaneously concentrating the AAV-containing iodixanol fraction using Amicon Ultra-15 filtration tubes (Merck; Darmstadt, Germany), the concentrate was sterile filtered.

    Transfection:

    Article Title: GFP-on mouse model for interrogation of in vivo gene editing.
    Article Snippet: .. 4 days after transfection, cells and media were collected using a cell scraper (Corning, Glendale, Arizona), pelleted by centrifugation for 2000×g for 10min, resuspended in 500μL hypertonic lysis buffer per plate (40mM Tris base, 500mM NaCl, 2mM MgCl2, 100U/mL salt-active nuclease (ArcticZymes; Wayne, PA)) and incubated at 37 °C for 1 h to lyse the cells. .. The media was decanted and combined with a 5x solution of 40% poly(ethylene glycol) 8000 (PEG8k) (Sigma-Aldrich, St. Louis,MO) in 2.5MNaCl (final concentration 8% PEG, 500mM NaCl).

    Lysis:

    Article Title: GFP-on mouse model for interrogation of in vivo gene editing.
    Article Snippet: .. 4 days after transfection, cells and media were collected using a cell scraper (Corning, Glendale, Arizona), pelleted by centrifugation for 2000×g for 10min, resuspended in 500μL hypertonic lysis buffer per plate (40mM Tris base, 500mM NaCl, 2mM MgCl2, 100U/mL salt-active nuclease (ArcticZymes; Wayne, PA)) and incubated at 37 °C for 1 h to lyse the cells. .. The media was decanted and combined with a 5x solution of 40% poly(ethylene glycol) 8000 (PEG8k) (Sigma-Aldrich, St. Louis,MO) in 2.5MNaCl (final concentration 8% PEG, 500mM NaCl).

    Fast Protein Liquid Chromatography:

    Article Title: VTA monosynaptic connections by local glutamate and GABA neurons and their distinct roles in behavior
    Article Snippet: .. Cell lysates were collected 40 h post-transfection, treated with SAN HQ nuclease (ArcticZymes Technologies ASA, Norway), and passed through an AVB columns (GE Healthcare, Silver Spring, MD) using a fast protein liquid chromatography (FPLC) machine (AKTA, Cytiva, Wilmington, DE). .. Purified viral particles were eluted from the column with sodium citrate (pH ~3.0) and titered using digital PCR (QIAcuity, QIAGEN, Germantown, MD) using a probe-based assay that recognizes the WPRE sequence. pAAV-nEF-Con/Foff-Gi-mCherry and pAAV-nEF-Coff/Fon-Gi-mCherry DREADDs were gifts from Karl Deisseroth (Addgene plasmid # 177673; RRID: Addgene_177673 and Addgene plasmid # 177669; RRID: Addgene_177669, respectively) and were packaged by the NIDA Genetic Engineering and Viral Vector Core (NIDA GEVV Core, Baltimore, MD), as described above.

    other:

    Article Title: Paralemmin-3 sustains the integrity of the lateral plasma membrane and subsurface cisternae of auditory hair cells
    Article Snippet: Viral particles from medium were precipitated with 40% polyethylene glycol 8000 (Acros Organics, Germany) in 500 mM NaCl for 2 h at 4 °C and then after centrifugation combined with cell pellets for processing.

    Plasmid Preparation:

    Article Title: AAV yield, bioactivity, and particle heterogeneity are impacted by genome size and non-coding DNA elements
    Article Snippet: .. Genomic DNA and remaining plasmid DNA were digested by incubation of the lysate with salt-active nuclease (ArcticZymes; Tromso, Norway) for 1 h, followed by polyethylene glycol (PEG) precipitation of proteins, including AAVs, for 3 h. The PEG pellet was resuspended overnight, and AAVs were purified by an iodixanol gradient to remove contaminating proteins and empty capsids. .. After exchanging the buffer and simultaneously concentrating the AAV-containing iodixanol fraction using Amicon Ultra-15 filtration tubes (Merck; Darmstadt, Germany), the concentrate was sterile filtered.



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    Image Search Results


    (A–C) Cyst severity is significantly reduced in vector-treated and hypertonic buffer-injected eyes at 1 month post injection (MPI). OCT images of vector-treated, hypertonic buffer-injected, and untreated eyes were collected at 1, 2, and 5 MPI. Volumetric scans were taken centering on the optic nerve. Representative OCT images demonstrate cyst changes at 1 MPI (A) . Cyst severity (B) was determined via manual segmentation of cyst area using the ImageJ software. Multiple comparisons were performed between every group at each time point. Non-significant comparisons were not annotated in the graphs. Vector-treated ( n = 10) or hypertonic buffer-injected Rs1 -KO eyes demonstrated significantly less cyst severity than untreated eyes (B) at 1 MPI. ONL thickness (C) remained stable from 1 to 5 MPI in all cohorts; however, at 5 MPI, vector treated eyes had statistically thinner ONLs than untreated eyes.

    Journal: Frontiers in Medicine

    Article Title: An osmolarity dependent mechanism partially ameliorates retinal cysts and rescues cone function in a mouse model of X-linked retinoschisis

    doi: 10.3389/fmed.2024.1302119

    Figure Lengend Snippet: (A–C) Cyst severity is significantly reduced in vector-treated and hypertonic buffer-injected eyes at 1 month post injection (MPI). OCT images of vector-treated, hypertonic buffer-injected, and untreated eyes were collected at 1, 2, and 5 MPI. Volumetric scans were taken centering on the optic nerve. Representative OCT images demonstrate cyst changes at 1 MPI (A) . Cyst severity (B) was determined via manual segmentation of cyst area using the ImageJ software. Multiple comparisons were performed between every group at each time point. Non-significant comparisons were not annotated in the graphs. Vector-treated ( n = 10) or hypertonic buffer-injected Rs1 -KO eyes demonstrated significantly less cyst severity than untreated eyes (B) at 1 MPI. ONL thickness (C) remained stable from 1 to 5 MPI in all cohorts; however, at 5 MPI, vector treated eyes had statistically thinner ONLs than untreated eyes.

    Article Snippet: Hypertonic injection buffer , 0.001% Gibco Pluronic F-68 Gibco dPBS (−/−) (Catalog #10010) (contains 155.2 mM NaCl) Additional 24.8 mM NaCl (for 180 mM NaCl total) pH 7.4.

    Techniques: Plasmid Preparation, Injection, Software

    (A–E) Hypertonic buffer robustly outperforms untreated and vector-treated eyes in light-adapted ERGs. Cone-dependent retina function was measured using two separate light-adapted ERG assays—the 5 Hz flicker (A) and 3.0 flash (B) . ERG amplitudes are shown from 1-month post injection (MPI) to 5 MPI for vector-treated, hypertonic buffer-injected, and untreated eyes in Rs1 -KO mice, and representative ERG traces at 5 MPI are illustrated. In response to both light-adapted assays, hypertonic buffer-injected eyes showed higher amplitudes than untreated and vector-treated eyes to 5 MPI (A) . The combined rod-cone function was measured by the 3.0 cd·s/m 2 standard combined response (SCR) under dark-adapted conditions (C) . At 1, 3, and 5-MPI, there was no difference between vector-treated, hypertonic buffer-injected, or untreated eyes. Isolated rod function was measured using 0.01 cd·s/m 2 dim flash after dark adaptation (D) . There was no persistent significant difference between hypertonic buffer-injected, isotonic buffer-injected, or untreated eyes in rod dominant ERG settings.

    Journal: Frontiers in Medicine

    Article Title: An osmolarity dependent mechanism partially ameliorates retinal cysts and rescues cone function in a mouse model of X-linked retinoschisis

    doi: 10.3389/fmed.2024.1302119

    Figure Lengend Snippet: (A–E) Hypertonic buffer robustly outperforms untreated and vector-treated eyes in light-adapted ERGs. Cone-dependent retina function was measured using two separate light-adapted ERG assays—the 5 Hz flicker (A) and 3.0 flash (B) . ERG amplitudes are shown from 1-month post injection (MPI) to 5 MPI for vector-treated, hypertonic buffer-injected, and untreated eyes in Rs1 -KO mice, and representative ERG traces at 5 MPI are illustrated. In response to both light-adapted assays, hypertonic buffer-injected eyes showed higher amplitudes than untreated and vector-treated eyes to 5 MPI (A) . The combined rod-cone function was measured by the 3.0 cd·s/m 2 standard combined response (SCR) under dark-adapted conditions (C) . At 1, 3, and 5-MPI, there was no difference between vector-treated, hypertonic buffer-injected, or untreated eyes. Isolated rod function was measured using 0.01 cd·s/m 2 dim flash after dark adaptation (D) . There was no persistent significant difference between hypertonic buffer-injected, isotonic buffer-injected, or untreated eyes in rod dominant ERG settings.

    Article Snippet: Hypertonic injection buffer , 0.001% Gibco Pluronic F-68 Gibco dPBS (−/−) (Catalog #10010) (contains 155.2 mM NaCl) Additional 24.8 mM NaCl (for 180 mM NaCl total) pH 7.4.

    Techniques: Plasmid Preparation, Injection, Isolation

    (A,B) Vector-treated and hypertonic buffer-injected Rs1 -KO mice perform faster in a light-adapted swim maze than untreated Rs1 -KO mice. Mice were placed in a swim maze and testing was performed under light-adapted (A) or dark-adapted (B) conditions. Time-to-platform was recorded. Each data point is representative of the average time-to-platform for a single mouse. In light-adapted conditions (untreated n = 8, vector-treated n = 4, hypertonic buffer n = 10.), we observed that vector-treated mice (2.92 s) and hypertonic buffer-injected mice (3.05 s) had a generally faster trend in average swim performance when finding a random platform than untreated mice (4.50 s) in lighted environments. In a dark-adapted environment, there is a less impactful effect on average time-to-platform between vector-treated (11.31 s), hypertonic buffer-injected (13.78 s), and untreated eyes (11.13 s), suggesting a limited effect on visual function in the dark (B) .

    Journal: Frontiers in Medicine

    Article Title: An osmolarity dependent mechanism partially ameliorates retinal cysts and rescues cone function in a mouse model of X-linked retinoschisis

    doi: 10.3389/fmed.2024.1302119

    Figure Lengend Snippet: (A,B) Vector-treated and hypertonic buffer-injected Rs1 -KO mice perform faster in a light-adapted swim maze than untreated Rs1 -KO mice. Mice were placed in a swim maze and testing was performed under light-adapted (A) or dark-adapted (B) conditions. Time-to-platform was recorded. Each data point is representative of the average time-to-platform for a single mouse. In light-adapted conditions (untreated n = 8, vector-treated n = 4, hypertonic buffer n = 10.), we observed that vector-treated mice (2.92 s) and hypertonic buffer-injected mice (3.05 s) had a generally faster trend in average swim performance when finding a random platform than untreated mice (4.50 s) in lighted environments. In a dark-adapted environment, there is a less impactful effect on average time-to-platform between vector-treated (11.31 s), hypertonic buffer-injected (13.78 s), and untreated eyes (11.13 s), suggesting a limited effect on visual function in the dark (B) .

    Article Snippet: Hypertonic injection buffer , 0.001% Gibco Pluronic F-68 Gibco dPBS (−/−) (Catalog #10010) (contains 155.2 mM NaCl) Additional 24.8 mM NaCl (for 180 mM NaCl total) pH 7.4.

    Techniques: Plasmid Preparation, Injection

    (A–C) Cyst area is reduced in hypertonic buffer and isotonic buffer-injected eyes, with greater reduction observed in the hypertonic buffer group. OCT images were collected at 2- and 3-weeks post injection (WPI), 1-month post-injection (MPI), and 2 MPI. Cyst area quantification (A) and outer nuclear layer (ONL) thickness (B) measurements were taken to assess cyst severity and photoreceptor cell survival, respectively. Representative OCT images demonstrate cysts at respective time points in all experimental groups (C) . Each data point is representative of one eye. At 3 WPI, there was less cyst severity in the hypertonic buffer-injected eyes compared to untreated eyes (A) . At 1 MPI, hypertonic and isotonic buffer-treated eyes both have less severe cysts than untreated and sham punctured eyes. This effect persisted until 2 MPI for the hypertonic buffer-injected eyes compared to untreated and sham punctured eyes. ONL thickness remains stable to 5 MPI and does not significantly differ between injection groups and untreated controls (B) .

    Journal: Frontiers in Medicine

    Article Title: An osmolarity dependent mechanism partially ameliorates retinal cysts and rescues cone function in a mouse model of X-linked retinoschisis

    doi: 10.3389/fmed.2024.1302119

    Figure Lengend Snippet: (A–C) Cyst area is reduced in hypertonic buffer and isotonic buffer-injected eyes, with greater reduction observed in the hypertonic buffer group. OCT images were collected at 2- and 3-weeks post injection (WPI), 1-month post-injection (MPI), and 2 MPI. Cyst area quantification (A) and outer nuclear layer (ONL) thickness (B) measurements were taken to assess cyst severity and photoreceptor cell survival, respectively. Representative OCT images demonstrate cysts at respective time points in all experimental groups (C) . Each data point is representative of one eye. At 3 WPI, there was less cyst severity in the hypertonic buffer-injected eyes compared to untreated eyes (A) . At 1 MPI, hypertonic and isotonic buffer-treated eyes both have less severe cysts than untreated and sham punctured eyes. This effect persisted until 2 MPI for the hypertonic buffer-injected eyes compared to untreated and sham punctured eyes. ONL thickness remains stable to 5 MPI and does not significantly differ between injection groups and untreated controls (B) .

    Article Snippet: Hypertonic injection buffer , 0.001% Gibco Pluronic F-68 Gibco dPBS (−/−) (Catalog #10010) (contains 155.2 mM NaCl) Additional 24.8 mM NaCl (for 180 mM NaCl total) pH 7.4.

    Techniques: Injection

    (A–E) Hypertonic buffer-injected eyes experience a robust rescue of cone functioning. Cone-dependent retina function was measured using two separate light-adapted ERG assays—the 3.0 cd·s/m 2 flash (A) , and the 5 Hz flicker (B) in hypertonic buffer, isotonic buffer, and sham punctured eyes in Rs1 -KO mice. Each data point represents one eye. ERG amplitudes are shown from 1-month post injection (MPI) to 5 MPI. In both light-adapted assays (A,B) , hypertonic and isotonic buffer-injected eyes showed higher amplitudes compared to untreated eyes at 1 MPI. Hypertonic buffer-injected eyes showed improvements over untreated and sham punctured eyes until 5 MPI. The combined rod-cone function was measured by the 3.0 cd·s/m 2 SCR under dark-adapted conditions (C) . Rod-dependent retina function was measured by the 0.01 dim flash (D) . In the 0.01 dim flash, hypertonic buffer-injected eyes showed improvements over untreated at 1 MPI and over both untreated and isotonic buffer-injected eyes at 2 MPI. However, significant differences over time were not observed.

    Journal: Frontiers in Medicine

    Article Title: An osmolarity dependent mechanism partially ameliorates retinal cysts and rescues cone function in a mouse model of X-linked retinoschisis

    doi: 10.3389/fmed.2024.1302119

    Figure Lengend Snippet: (A–E) Hypertonic buffer-injected eyes experience a robust rescue of cone functioning. Cone-dependent retina function was measured using two separate light-adapted ERG assays—the 3.0 cd·s/m 2 flash (A) , and the 5 Hz flicker (B) in hypertonic buffer, isotonic buffer, and sham punctured eyes in Rs1 -KO mice. Each data point represents one eye. ERG amplitudes are shown from 1-month post injection (MPI) to 5 MPI. In both light-adapted assays (A,B) , hypertonic and isotonic buffer-injected eyes showed higher amplitudes compared to untreated eyes at 1 MPI. Hypertonic buffer-injected eyes showed improvements over untreated and sham punctured eyes until 5 MPI. The combined rod-cone function was measured by the 3.0 cd·s/m 2 SCR under dark-adapted conditions (C) . Rod-dependent retina function was measured by the 0.01 dim flash (D) . In the 0.01 dim flash, hypertonic buffer-injected eyes showed improvements over untreated at 1 MPI and over both untreated and isotonic buffer-injected eyes at 2 MPI. However, significant differences over time were not observed.

    Article Snippet: Hypertonic injection buffer , 0.001% Gibco Pluronic F-68 Gibco dPBS (−/−) (Catalog #10010) (contains 155.2 mM NaCl) Additional 24.8 mM NaCl (for 180 mM NaCl total) pH 7.4.

    Techniques: Injection

    (A–F) Peak cyst severity on OCT is negatively associated with cone function on ERG at 5 MPI. Peak cyst severity of hypertonic buffer-injected, isotonic buffer-injected, sham-punctured and untreated eyes was determined by comparing the cyst area measurements of each eye at 1, 2, and 5 MPI. The cyst area of each eye representing their peak severity was correlated with its ERG amplitudes at 5 MPI. Each dot represents one eye. A negative correlation was found between cyst area and ERG amplitudes in the light-adapted 5 Hz flicker and 3.0 cd·s/m 2 flash (A,B) . Little to no correlation was found between peak cyst area and ERG amplitudes in dark-adapted 0.01 cd·s/m 2 or 3.0 cd·s/m 2 SCR (C–F) . These findings suggest increasing cyst area has greater impact on impairing cone electrical function, than rod photoreceptors.

    Journal: Frontiers in Medicine

    Article Title: An osmolarity dependent mechanism partially ameliorates retinal cysts and rescues cone function in a mouse model of X-linked retinoschisis

    doi: 10.3389/fmed.2024.1302119

    Figure Lengend Snippet: (A–F) Peak cyst severity on OCT is negatively associated with cone function on ERG at 5 MPI. Peak cyst severity of hypertonic buffer-injected, isotonic buffer-injected, sham-punctured and untreated eyes was determined by comparing the cyst area measurements of each eye at 1, 2, and 5 MPI. The cyst area of each eye representing their peak severity was correlated with its ERG amplitudes at 5 MPI. Each dot represents one eye. A negative correlation was found between cyst area and ERG amplitudes in the light-adapted 5 Hz flicker and 3.0 cd·s/m 2 flash (A,B) . Little to no correlation was found between peak cyst area and ERG amplitudes in dark-adapted 0.01 cd·s/m 2 or 3.0 cd·s/m 2 SCR (C–F) . These findings suggest increasing cyst area has greater impact on impairing cone electrical function, than rod photoreceptors.

    Article Snippet: Hypertonic injection buffer , 0.001% Gibco Pluronic F-68 Gibco dPBS (−/−) (Catalog #10010) (contains 155.2 mM NaCl) Additional 24.8 mM NaCl (for 180 mM NaCl total) pH 7.4.

    Techniques: Injection

    (A,B) Increased cone density at 5 months after subretinal injections of a hypertonic or an isotonic buffer. Retinal sections were collected from untreated Rs1 -KO eyes, hypertonic buffer-injected eyes, isotonic buffer-injected eyes, and sham puncture eyes at approximately 6-months of age. Sections were stained by PNA to visualize cone outer segments. The number of cone outer segments per 100 μm of the retina was quantified by three participants masked to treatment groups, and quantifications by these three participants were averaged. Representative immunofluorescent staining (red: PNA) of cone outer segments (arrow), demonstrating increased density of cones in hypertonic buffer-injected and isotonic buffer-injected eyes compared to untreated eyes with sparse cone staining (A) . Synapses were visualized by an anti-RIBEYE antibody (green). Each data point represents the averaged cone density of one eye. Hypertonic buffer-injected eyes and isotonic buffer-injected eyes show significantly higher cone density per 100 μm than untreated Rs1 -KO eyes (B) .

    Journal: Frontiers in Medicine

    Article Title: An osmolarity dependent mechanism partially ameliorates retinal cysts and rescues cone function in a mouse model of X-linked retinoschisis

    doi: 10.3389/fmed.2024.1302119

    Figure Lengend Snippet: (A,B) Increased cone density at 5 months after subretinal injections of a hypertonic or an isotonic buffer. Retinal sections were collected from untreated Rs1 -KO eyes, hypertonic buffer-injected eyes, isotonic buffer-injected eyes, and sham puncture eyes at approximately 6-months of age. Sections were stained by PNA to visualize cone outer segments. The number of cone outer segments per 100 μm of the retina was quantified by three participants masked to treatment groups, and quantifications by these three participants were averaged. Representative immunofluorescent staining (red: PNA) of cone outer segments (arrow), demonstrating increased density of cones in hypertonic buffer-injected and isotonic buffer-injected eyes compared to untreated eyes with sparse cone staining (A) . Synapses were visualized by an anti-RIBEYE antibody (green). Each data point represents the averaged cone density of one eye. Hypertonic buffer-injected eyes and isotonic buffer-injected eyes show significantly higher cone density per 100 μm than untreated Rs1 -KO eyes (B) .

    Article Snippet: Hypertonic injection buffer , 0.001% Gibco Pluronic F-68 Gibco dPBS (−/−) (Catalog #10010) (contains 155.2 mM NaCl) Additional 24.8 mM NaCl (for 180 mM NaCl total) pH 7.4.

    Techniques: Injection, Staining