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  • 99
    Thermo Fisher fluo 4
    BrainPhys Imaging optimally supports fast-rising calcium spikes in human neurons in vitro. Intracellular Ca 2+ changes in human PSC-derived neurons were measured with time-lapse image sequences of a Ca 2+ sensor <t>(Fluo-4</t> AM). A total 1225 human neurons were analyzed across thirteen fields of view (FOVs) from four coverslips ( n = 4 biologically independent experiments). Regions of interest (ROIs) were drawn on the cell soma to determine fluorescence intensity changes (Δ F / F 0 ) over time. Active events (Δ F / F 0 > 5% from baseline) were manually categorized into fast-rising Ca 2+ spikes or slow-rising Ca 2+ waves (see Fig. S8A ). The same FOVs were imaged in artificial cerebrospinal fluid (ACSF), BPI, FluoroBrite, and NEUMO. a Example Fluo-4 AM fluorescence image of a neuronal population in BPI. White circles represent active ROIs. Fluorescent intensity represents intracellular Ca 2+ levels. b Typical fast-rising Ca 2+ spikes from five active cells in BPI. c , d No significant difference was observed between the average properties of fast-rising unitary Ca 2+ spike events in BPI ( n = 16 cells) and ACSF ( n = 24 cells) from six FOVs across two coverslips. e , g No significant difference was observed between the proportions of cells with fast Ca 2+ spikes or slow Ca 2+ waves in BPI and ACSF. The total active cells (BPI: n = 329; ACSF n = 308) were compared across six FOVs from two coverslips. Voltage-gated sodium channel blocker Tetrodotoxin (TTX, 1 µM) significantly reduced Ca 2+ spikes and waves. See also Supplementary Fig. 8a . i , k Comparison of Ca 2+ signals in BPI, NEUMO, and FluoroBrite show significant differences in the proportions of cells with Ca 2+ spikes but not waves. Active cells with Ca 2+ spikes and spikes/waves in BPI ( n = 243 cells), FluoroBrite ( n = 39 cells) and NEUMO ( n = 18 cells) or with Ca 2+ waves in BPI ( n = 288 cells), FluoroBrite ( n = 302 cells), and NEUMO ( n = 356 cells) were compared across seven FOVs from two coverslips. f , h , j , l Example traces from the same ROIs in different media. Symbols in c – e , g , i , k represent cells recorded first (triangles), second (circles), third (square), fourth (diamond) in either medium. Values are presented as mean ± SEM. Significance in c – e , g , i , k determined via two-tailed nonparametric unpaired (Mann–Whitney) tests. ns, P > 0.05.
    Fluo 4, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 8017 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore fluo 4
    The CDPK4/PKG functional interaction is conserved to control P. falciparum merozoite invasion. a Difference in ring and schizont parasitaemia after 3 h of C2 treatment added on synchronised segmented schizonts in the control 3D7, PKG T618Q , and CDPK4-KO lines (error bars show standard deviations from the mean; 2 independent cultures; two-tailed t -test). b Difference in ring and schizont parasitaemia after 3 h of C2 treatment added on synchronised segmented schizonts in the CDPK4-KO line and in daughter lines complemented with P. berghei CDPK4 or CDPK4 S147M alleles (error bars show standard deviations from the mean; 2 independent cultures; two-tailed t -test). c Difference in ring and schizont parasitaemia after 3 h of Compound A treatment added on synchronised segmented schizonts in the control 3D7, PKG T618Q , and CDPK4-KO lines (error bars show standard deviations from the mean; 2 independent cultures; two-tailed t -test). d Determination of the ATP Km for recombinant PfPKG and PfPKG T618Q . e Fluorescence of the cell permeable calcium probe <t>Fluo-4-AM</t> in non-synchronised 3D7 and PKG T618Q parasites labelled with the DNA dye Vybrant Green (error bars show standard deviations from the mean; 2 biological replicates). f Relative fluorescence response of Pf3D7 and PfPKG T618Q lines loaded with the calcium indicator Fluo-4-AM in response to the phosphodiesterase inhibitors Zaprinast and BIPPO as well as the ionophore A23187 (error bars show standard deviations from the mean; 3 or 4 biological replicates, two-tailed t -test)
    Fluo 4, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 249 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher fluo 4 nw calcium assay kit
    Ca 2+ influx through 2B channels is enhanced by partial blockade of the inhibitory activity in the spinal cord. ( A ) Fluorescence imaging of Ca 2+ using the Ca 2+ -sensitive dye <t>fluo-4.</t> Scale bar = 250 µm. ( B ) Time course of Ca 2+ influx calculated from the fluorescence intensity at the indicated regions of interest. Calibration, Δ F / F 0 2%, 100 ms. ( C ) Ca 2+ response (Δ F / F 0 ) (1.00 ± 0.18 in control at 7 DIV, n = 23, Ns = 23, Nm = 14; 0.46 ± 0.06 in control at 12–15 DIV, n = 12, Ns = 12, Nm = 12; 1.15 ± 0.23 in strychnine, n = 18, Ns = 18, Nm = 16). * p
    Fluo 4 Nw Calcium Assay Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 749 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher fluo 4 acetoxymethyl ester
    Ca 2+ influx through 2B channels is enhanced by partial blockade of the inhibitory activity in the spinal cord. ( A ) Fluorescence imaging of Ca 2+ using the Ca 2+ -sensitive dye <t>fluo-4.</t> Scale bar = 250 µm. ( B ) Time course of Ca 2+ influx calculated from the fluorescence intensity at the indicated regions of interest. Calibration, Δ F / F 0 2%, 100 ms. ( C ) Ca 2+ response (Δ F / F 0 ) (1.00 ± 0.18 in control at 7 DIV, n = 23, Ns = 23, Nm = 14; 0.46 ± 0.06 in control at 12–15 DIV, n = 12, Ns = 12, Nm = 12; 1.15 ± 0.23 in strychnine, n = 18, Ns = 18, Nm = 16). * p
    Fluo 4 Acetoxymethyl Ester, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 709 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher fluo 4 direct calcium assay kit
    Organization and maturation of cerebral cortical neurons a, Immunohistochemical staining at day 30 showing preplate (Tbr1) with early signs of radial organization (MAP2, bracket i) and the presence of an IZ-like layer (bracket ii) adjacent to the VZ/SVZ (bracket iii). DAPI marks nuclei (blue). b, Reelin staining indicating Cajal-Retzius cells along the basal surface of dorsal cortical tissue. c, Staining for early born (Ctip2) and late born (Satb2) neurons at 75 days differentiation reveals separation and rudimentary inside-out organization. d. False color heat map frames from <t>Fluo-4</t> calcium live imaging revealing spontaneous calcium surges (arrowheads). Time is displayed in min:sec. e. Single cell tracings of calcium surges with glutamate application (regions of interest, ROI, outlined in left panel) as measured by change in fluorescence (arbitrary units). Arrows mark the time of addition of glutamate. f. Single cell tracing (ROIs marked in image at left) of calcium surges before (left panels) and after the addition of TTX (right panels). Scale bars: 100 μm.
    Fluo 4 Direct Calcium Assay Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 492 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher ca2 indicator fluo 4
    Organization and maturation of cerebral cortical neurons a, Immunohistochemical staining at day 30 showing preplate (Tbr1) with early signs of radial organization (MAP2, bracket i) and the presence of an IZ-like layer (bracket ii) adjacent to the VZ/SVZ (bracket iii). DAPI marks nuclei (blue). b, Reelin staining indicating Cajal-Retzius cells along the basal surface of dorsal cortical tissue. c, Staining for early born (Ctip2) and late born (Satb2) neurons at 75 days differentiation reveals separation and rudimentary inside-out organization. d. False color heat map frames from <t>Fluo-4</t> calcium live imaging revealing spontaneous calcium surges (arrowheads). Time is displayed in min:sec. e. Single cell tracings of calcium surges with glutamate application (regions of interest, ROI, outlined in left panel) as measured by change in fluorescence (arbitrary units). Arrows mark the time of addition of glutamate. f. Single cell tracing (ROIs marked in image at left) of calcium surges before (left panels) and after the addition of TTX (right panels). Scale bars: 100 μm.
    Ca2 Indicator Fluo 4, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 475 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Dojindo Labs fluo 4 am
    Comparison of the influence of the cell lysis solution and pluronic F-127 on <t>fluo-4/AM</t> dyeing. Fluorescence intensity using cell lysis solution as an auxiliary reagent was significantly higher ( P
    Fluo 4 Am, supplied by Dojindo Labs, used in various techniques. Bioz Stars score: 93/100, based on 249 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    96
    Thermo Fisher fluo 4 pentapotassium salt
    Comparison of the influence of the cell lysis solution and pluronic F-127 on <t>fluo-4/AM</t> dyeing. Fluorescence intensity using cell lysis solution as an auxiliary reagent was significantly higher ( P
    Fluo 4 Pentapotassium Salt, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 96/100, based on 166 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Olympus fluo 4
    Cycling of Ca 2+ in CPCs in response to electrical stimulation is dependent on the LTCC, IP 3 R, and external Ca 2+ . (a) Representative Ca 2+ oscillations recorded from <t>fluo-4/AM-loaded</t> CPCs at 0.5 Hz electrical stimulation in 2 mM Ca 2+ Tyrode's solution, pretreated with nifedipine or 2-APB to inhibit the LTCC or IP 3 R, respectively, or in 0 Ca 2+ Tyrode's solution. Traces have been aligned in time based on when electrical stimulation was applied for comparison. Arrow indicates the start of stimulation. (b) Summary bar graph of Ca 2+ oscillation amplitudes under the various recording conditions in (a). Data represent mean ± SD. ∗ p
    Fluo 4, supplied by Olympus, used in various techniques. Bioz Stars score: 92/100, based on 282 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    97
    Thermo Fisher mag fluo 4
    Δ F/F R responses measured from a midshipman swimbladder fiber that was injected with <t>Fluo-4</t> and excited by a 67-Hz stimulus train that lasted 10 s. (A) The response during the first part of the recording, which included 0.05 s of baseline plus 0.45 s of stimulation. (B) The response during the last part of the recording (9–9.5 s after onset of stimulation).
    Mag Fluo 4, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 97/100, based on 96 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher fluo 4 calcium imaging kit
    Δ F/F R responses measured from a midshipman swimbladder fiber that was injected with <t>Fluo-4</t> and excited by a 67-Hz stimulus train that lasted 10 s. (A) The response during the first part of the recording, which included 0.05 s of baseline plus 0.45 s of stimulation. (B) The response during the last part of the recording (9–9.5 s after onset of stimulation).
    Fluo 4 Calcium Imaging Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 52 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    BrainPhys Imaging optimally supports fast-rising calcium spikes in human neurons in vitro. Intracellular Ca 2+ changes in human PSC-derived neurons were measured with time-lapse image sequences of a Ca 2+ sensor (Fluo-4 AM). A total 1225 human neurons were analyzed across thirteen fields of view (FOVs) from four coverslips ( n = 4 biologically independent experiments). Regions of interest (ROIs) were drawn on the cell soma to determine fluorescence intensity changes (Δ F / F 0 ) over time. Active events (Δ F / F 0 > 5% from baseline) were manually categorized into fast-rising Ca 2+ spikes or slow-rising Ca 2+ waves (see Fig. S8A ). The same FOVs were imaged in artificial cerebrospinal fluid (ACSF), BPI, FluoroBrite, and NEUMO. a Example Fluo-4 AM fluorescence image of a neuronal population in BPI. White circles represent active ROIs. Fluorescent intensity represents intracellular Ca 2+ levels. b Typical fast-rising Ca 2+ spikes from five active cells in BPI. c , d No significant difference was observed between the average properties of fast-rising unitary Ca 2+ spike events in BPI ( n = 16 cells) and ACSF ( n = 24 cells) from six FOVs across two coverslips. e , g No significant difference was observed between the proportions of cells with fast Ca 2+ spikes or slow Ca 2+ waves in BPI and ACSF. The total active cells (BPI: n = 329; ACSF n = 308) were compared across six FOVs from two coverslips. Voltage-gated sodium channel blocker Tetrodotoxin (TTX, 1 µM) significantly reduced Ca 2+ spikes and waves. See also Supplementary Fig. 8a . i , k Comparison of Ca 2+ signals in BPI, NEUMO, and FluoroBrite show significant differences in the proportions of cells with Ca 2+ spikes but not waves. Active cells with Ca 2+ spikes and spikes/waves in BPI ( n = 243 cells), FluoroBrite ( n = 39 cells) and NEUMO ( n = 18 cells) or with Ca 2+ waves in BPI ( n = 288 cells), FluoroBrite ( n = 302 cells), and NEUMO ( n = 356 cells) were compared across seven FOVs from two coverslips. f , h , j , l Example traces from the same ROIs in different media. Symbols in c – e , g , i , k represent cells recorded first (triangles), second (circles), third (square), fourth (diamond) in either medium. Values are presented as mean ± SEM. Significance in c – e , g , i , k determined via two-tailed nonparametric unpaired (Mann–Whitney) tests. ns, P > 0.05.

    Journal: Nature Communications

    Article Title: BrainPhys neuronal medium optimized for imaging and optogenetics in vitro

    doi: 10.1038/s41467-020-19275-x

    Figure Lengend Snippet: BrainPhys Imaging optimally supports fast-rising calcium spikes in human neurons in vitro. Intracellular Ca 2+ changes in human PSC-derived neurons were measured with time-lapse image sequences of a Ca 2+ sensor (Fluo-4 AM). A total 1225 human neurons were analyzed across thirteen fields of view (FOVs) from four coverslips ( n = 4 biologically independent experiments). Regions of interest (ROIs) were drawn on the cell soma to determine fluorescence intensity changes (Δ F / F 0 ) over time. Active events (Δ F / F 0 > 5% from baseline) were manually categorized into fast-rising Ca 2+ spikes or slow-rising Ca 2+ waves (see Fig. S8A ). The same FOVs were imaged in artificial cerebrospinal fluid (ACSF), BPI, FluoroBrite, and NEUMO. a Example Fluo-4 AM fluorescence image of a neuronal population in BPI. White circles represent active ROIs. Fluorescent intensity represents intracellular Ca 2+ levels. b Typical fast-rising Ca 2+ spikes from five active cells in BPI. c , d No significant difference was observed between the average properties of fast-rising unitary Ca 2+ spike events in BPI ( n = 16 cells) and ACSF ( n = 24 cells) from six FOVs across two coverslips. e , g No significant difference was observed between the proportions of cells with fast Ca 2+ spikes or slow Ca 2+ waves in BPI and ACSF. The total active cells (BPI: n = 329; ACSF n = 308) were compared across six FOVs from two coverslips. Voltage-gated sodium channel blocker Tetrodotoxin (TTX, 1 µM) significantly reduced Ca 2+ spikes and waves. See also Supplementary Fig. 8a . i , k Comparison of Ca 2+ signals in BPI, NEUMO, and FluoroBrite show significant differences in the proportions of cells with Ca 2+ spikes but not waves. Active cells with Ca 2+ spikes and spikes/waves in BPI ( n = 243 cells), FluoroBrite ( n = 39 cells) and NEUMO ( n = 18 cells) or with Ca 2+ waves in BPI ( n = 288 cells), FluoroBrite ( n = 302 cells), and NEUMO ( n = 356 cells) were compared across seven FOVs from two coverslips. f , h , j , l Example traces from the same ROIs in different media. Symbols in c – e , g , i , k represent cells recorded first (triangles), second (circles), third (square), fourth (diamond) in either medium. Values are presented as mean ± SEM. Significance in c – e , g , i , k determined via two-tailed nonparametric unpaired (Mann–Whitney) tests. ns, P > 0.05.

    Article Snippet: Calcium imaging hPSC-derived neurons attached onto coverslips in 48-well plates were incubated with a 0.5 μl of Fluo4-AM(1 mM; Cat. No. F14201, Thermo Fisher Scientific) in 500 μl of neuromedium, per well (final Fluo-4 AM concentration in media: 1 μM).

    Techniques: Imaging, In Vitro, Derivative Assay, Fluorescence, Two Tailed Test, MANN-WHITNEY

    The CDPK4/PKG functional interaction is conserved to control P. falciparum merozoite invasion. a Difference in ring and schizont parasitaemia after 3 h of C2 treatment added on synchronised segmented schizonts in the control 3D7, PKG T618Q , and CDPK4-KO lines (error bars show standard deviations from the mean; 2 independent cultures; two-tailed t -test). b Difference in ring and schizont parasitaemia after 3 h of C2 treatment added on synchronised segmented schizonts in the CDPK4-KO line and in daughter lines complemented with P. berghei CDPK4 or CDPK4 S147M alleles (error bars show standard deviations from the mean; 2 independent cultures; two-tailed t -test). c Difference in ring and schizont parasitaemia after 3 h of Compound A treatment added on synchronised segmented schizonts in the control 3D7, PKG T618Q , and CDPK4-KO lines (error bars show standard deviations from the mean; 2 independent cultures; two-tailed t -test). d Determination of the ATP Km for recombinant PfPKG and PfPKG T618Q . e Fluorescence of the cell permeable calcium probe Fluo-4-AM in non-synchronised 3D7 and PKG T618Q parasites labelled with the DNA dye Vybrant Green (error bars show standard deviations from the mean; 2 biological replicates). f Relative fluorescence response of Pf3D7 and PfPKG T618Q lines loaded with the calcium indicator Fluo-4-AM in response to the phosphodiesterase inhibitors Zaprinast and BIPPO as well as the ionophore A23187 (error bars show standard deviations from the mean; 3 or 4 biological replicates, two-tailed t -test)

    Journal: Nature Communications

    Article Title: Epistasis studies reveal redundancy among calcium-dependent protein kinases in motility and invasion of malaria parasites

    doi: 10.1038/s41467-018-06733-w

    Figure Lengend Snippet: The CDPK4/PKG functional interaction is conserved to control P. falciparum merozoite invasion. a Difference in ring and schizont parasitaemia after 3 h of C2 treatment added on synchronised segmented schizonts in the control 3D7, PKG T618Q , and CDPK4-KO lines (error bars show standard deviations from the mean; 2 independent cultures; two-tailed t -test). b Difference in ring and schizont parasitaemia after 3 h of C2 treatment added on synchronised segmented schizonts in the CDPK4-KO line and in daughter lines complemented with P. berghei CDPK4 or CDPK4 S147M alleles (error bars show standard deviations from the mean; 2 independent cultures; two-tailed t -test). c Difference in ring and schizont parasitaemia after 3 h of Compound A treatment added on synchronised segmented schizonts in the control 3D7, PKG T618Q , and CDPK4-KO lines (error bars show standard deviations from the mean; 2 independent cultures; two-tailed t -test). d Determination of the ATP Km for recombinant PfPKG and PfPKG T618Q . e Fluorescence of the cell permeable calcium probe Fluo-4-AM in non-synchronised 3D7 and PKG T618Q parasites labelled with the DNA dye Vybrant Green (error bars show standard deviations from the mean; 2 biological replicates). f Relative fluorescence response of Pf3D7 and PfPKG T618Q lines loaded with the calcium indicator Fluo-4-AM in response to the phosphodiesterase inhibitors Zaprinast and BIPPO as well as the ionophore A23187 (error bars show standard deviations from the mean; 3 or 4 biological replicates, two-tailed t -test)

    Article Snippet: Changes in the levels of intracellular free calcium were measured using Fluo-4 (Sigma) loaded P. falciparum late-stage schizonts.

    Techniques: Functional Assay, Two Tailed Test, Recombinant, Fluorescence

    Ca 2+ influx through 2B channels is enhanced by partial blockade of the inhibitory activity in the spinal cord. ( A ) Fluorescence imaging of Ca 2+ using the Ca 2+ -sensitive dye fluo-4. Scale bar = 250 µm. ( B ) Time course of Ca 2+ influx calculated from the fluorescence intensity at the indicated regions of interest. Calibration, Δ F / F 0 2%, 100 ms. ( C ) Ca 2+ response (Δ F / F 0 ) (1.00 ± 0.18 in control at 7 DIV, n = 23, Ns = 23, Nm = 14; 0.46 ± 0.06 in control at 12–15 DIV, n = 12, Ns = 12, Nm = 12; 1.15 ± 0.23 in strychnine, n = 18, Ns = 18, Nm = 16). * p

    Journal: Scientific Reports

    Article Title: The decline in synaptic GluN2B and rise in inhibitory neurotransmission determine the end of a critical period

    doi: 10.1038/srep34196

    Figure Lengend Snippet: Ca 2+ influx through 2B channels is enhanced by partial blockade of the inhibitory activity in the spinal cord. ( A ) Fluorescence imaging of Ca 2+ using the Ca 2+ -sensitive dye fluo-4. Scale bar = 250 µm. ( B ) Time course of Ca 2+ influx calculated from the fluorescence intensity at the indicated regions of interest. Calibration, Δ F / F 0 2%, 100 ms. ( C ) Ca 2+ response (Δ F / F 0 ) (1.00 ± 0.18 in control at 7 DIV, n = 23, Ns = 23, Nm = 14; 0.46 ± 0.06 in control at 12–15 DIV, n = 12, Ns = 12, Nm = 12; 1.15 ± 0.23 in strychnine, n = 18, Ns = 18, Nm = 16). * p

    Article Snippet: Calcium imaging Slices were incubated with the membrane permeant AM ester of the Ca2+ -sensitive dye fluo-4 (5 μg/ml; Molecular Probes) and 0.04% pluronic acid for 90 min at 37 °C, after which optical measurements of fluorescence changes were made using a high-speed camera system (MiCAM02).

    Techniques: Activity Assay, Fluorescence, Imaging, Mass Spectrometry

    Organization and maturation of cerebral cortical neurons a, Immunohistochemical staining at day 30 showing preplate (Tbr1) with early signs of radial organization (MAP2, bracket i) and the presence of an IZ-like layer (bracket ii) adjacent to the VZ/SVZ (bracket iii). DAPI marks nuclei (blue). b, Reelin staining indicating Cajal-Retzius cells along the basal surface of dorsal cortical tissue. c, Staining for early born (Ctip2) and late born (Satb2) neurons at 75 days differentiation reveals separation and rudimentary inside-out organization. d. False color heat map frames from Fluo-4 calcium live imaging revealing spontaneous calcium surges (arrowheads). Time is displayed in min:sec. e. Single cell tracings of calcium surges with glutamate application (regions of interest, ROI, outlined in left panel) as measured by change in fluorescence (arbitrary units). Arrows mark the time of addition of glutamate. f. Single cell tracing (ROIs marked in image at left) of calcium surges before (left panels) and after the addition of TTX (right panels). Scale bars: 100 μm.

    Journal: Nature

    Article Title: Cerebral organoids model human brain development and microcephaly

    doi: 10.1038/nature12517

    Figure Lengend Snippet: Organization and maturation of cerebral cortical neurons a, Immunohistochemical staining at day 30 showing preplate (Tbr1) with early signs of radial organization (MAP2, bracket i) and the presence of an IZ-like layer (bracket ii) adjacent to the VZ/SVZ (bracket iii). DAPI marks nuclei (blue). b, Reelin staining indicating Cajal-Retzius cells along the basal surface of dorsal cortical tissue. c, Staining for early born (Ctip2) and late born (Satb2) neurons at 75 days differentiation reveals separation and rudimentary inside-out organization. d. False color heat map frames from Fluo-4 calcium live imaging revealing spontaneous calcium surges (arrowheads). Time is displayed in min:sec. e. Single cell tracings of calcium surges with glutamate application (regions of interest, ROI, outlined in left panel) as measured by change in fluorescence (arbitrary units). Arrows mark the time of addition of glutamate. f. Single cell tracing (ROIs marked in image at left) of calcium surges before (left panels) and after the addition of TTX (right panels). Scale bars: 100 μm.

    Article Snippet: For calcium imaging, Fluo-4 direct (Life Technologies) was prepared according to manufacturer and applied 60 min before the start of imaging.

    Techniques: Immunohistochemistry, Staining, Imaging, Size-exclusion Chromatography, Fluorescence

    Comparison of the influence of the cell lysis solution and pluronic F-127 on fluo-4/AM dyeing. Fluorescence intensity using cell lysis solution as an auxiliary reagent was significantly higher ( P

    Journal: PLoS ONE

    Article Title: Rapid and Inexpensive Method of Loading Fluorescent Dye into Pollen Tubes and Root Hairs

    doi: 10.1371/journal.pone.0152320

    Figure Lengend Snippet: Comparison of the influence of the cell lysis solution and pluronic F-127 on fluo-4/AM dyeing. Fluorescence intensity using cell lysis solution as an auxiliary reagent was significantly higher ( P

    Article Snippet: Pollen tube fluorescence was developed and detected using the following steps: Exactly 50 μg of fluo-4/AM was dissolved in 91.2 μL of anhydrous dimethylsulfoxide (Dojindo) in a microcentrifuge tube.

    Techniques: Lysis, Fluorescence

    Loading of fluo-4/AM into pollen tubes under different conditions. (a, b) Control pollen tube that was not loaded with fluo-4/AM. (c-h) Pollen tubes loaded with fluo-4/AM along with cell lysis solution for 5 min (c, d), 15 min (e, f) and 30 min (g, h). (i, j) Pollen tubes loaded for 30 min without cell lysis solution.

    Journal: PLoS ONE

    Article Title: Rapid and Inexpensive Method of Loading Fluorescent Dye into Pollen Tubes and Root Hairs

    doi: 10.1371/journal.pone.0152320

    Figure Lengend Snippet: Loading of fluo-4/AM into pollen tubes under different conditions. (a, b) Control pollen tube that was not loaded with fluo-4/AM. (c-h) Pollen tubes loaded with fluo-4/AM along with cell lysis solution for 5 min (c, d), 15 min (e, f) and 30 min (g, h). (i, j) Pollen tubes loaded for 30 min without cell lysis solution.

    Article Snippet: Pollen tube fluorescence was developed and detected using the following steps: Exactly 50 μg of fluo-4/AM was dissolved in 91.2 μL of anhydrous dimethylsulfoxide (Dojindo) in a microcentrifuge tube.

    Techniques: Lysis

    Effect of EGTA on the Ca 2+ gradient in the pollen tube apical regions. (a) Pollen tube loaded with fluo-4/AM without EGTA (control). (b) Pollen tube after addition of 1 mM EGTA following fluo-4/AM loading. (c) Pollen tube after addition of 1 mM EGTA during fluo-4/AM loading. Scale bars: 100 μm (a), 60 μm (b) and 30 μm (c).

    Journal: PLoS ONE

    Article Title: Rapid and Inexpensive Method of Loading Fluorescent Dye into Pollen Tubes and Root Hairs

    doi: 10.1371/journal.pone.0152320

    Figure Lengend Snippet: Effect of EGTA on the Ca 2+ gradient in the pollen tube apical regions. (a) Pollen tube loaded with fluo-4/AM without EGTA (control). (b) Pollen tube after addition of 1 mM EGTA following fluo-4/AM loading. (c) Pollen tube after addition of 1 mM EGTA during fluo-4/AM loading. Scale bars: 100 μm (a), 60 μm (b) and 30 μm (c).

    Article Snippet: Pollen tube fluorescence was developed and detected using the following steps: Exactly 50 μg of fluo-4/AM was dissolved in 91.2 μL of anhydrous dimethylsulfoxide (Dojindo) in a microcentrifuge tube.

    Techniques:

    Loading fluo-4/AM into the root hair and pollen tube of Arabidopsis . (a) Control root hair that was not loaded with fluo-4/AM. (b) Root hair loaded with fluo-4/AM along with cell lysis solution for 15 min. (c-d) Pollen tubes loaded with fluo-4/AM along with cell lysis solution for 15 min. Note: The root hairs were barely observable in the fluorescence microscopy image because the autofluorescence of the root hairs was very weak. To see the root hairs clearly in the image, we improved the overall image brightness with Photoshop software. Therefore, the background of image (a) is different from the background of the other images (b, c and d).

    Journal: PLoS ONE

    Article Title: Rapid and Inexpensive Method of Loading Fluorescent Dye into Pollen Tubes and Root Hairs

    doi: 10.1371/journal.pone.0152320

    Figure Lengend Snippet: Loading fluo-4/AM into the root hair and pollen tube of Arabidopsis . (a) Control root hair that was not loaded with fluo-4/AM. (b) Root hair loaded with fluo-4/AM along with cell lysis solution for 15 min. (c-d) Pollen tubes loaded with fluo-4/AM along with cell lysis solution for 15 min. Note: The root hairs were barely observable in the fluorescence microscopy image because the autofluorescence of the root hairs was very weak. To see the root hairs clearly in the image, we improved the overall image brightness with Photoshop software. Therefore, the background of image (a) is different from the background of the other images (b, c and d).

    Article Snippet: Pollen tube fluorescence was developed and detected using the following steps: Exactly 50 μg of fluo-4/AM was dissolved in 91.2 μL of anhydrous dimethylsulfoxide (Dojindo) in a microcentrifuge tube.

    Techniques: Lysis, Fluorescence, Microscopy, Software

    Effect of different concentrations of La 3+ on the calcium gradient in the pollen tube apical regions. (a) Pollen tube loaded with fluo-4/AM without La 3+ (control). Pollen tubes after loading with fluo-4/AM followed by the addition of 10 μM La 3+ (b), 100 μM La 3+ (c), and 1 mM La 3+ (d). (e) Pollen tube after loading with fluo-4/AM in the presence of 1 mM La 3+ . Scale bars: 100 μm (a), 90 μm (b and c), 60 μm (d) and 70 μm (e).

    Journal: PLoS ONE

    Article Title: Rapid and Inexpensive Method of Loading Fluorescent Dye into Pollen Tubes and Root Hairs

    doi: 10.1371/journal.pone.0152320

    Figure Lengend Snippet: Effect of different concentrations of La 3+ on the calcium gradient in the pollen tube apical regions. (a) Pollen tube loaded with fluo-4/AM without La 3+ (control). Pollen tubes after loading with fluo-4/AM followed by the addition of 10 μM La 3+ (b), 100 μM La 3+ (c), and 1 mM La 3+ (d). (e) Pollen tube after loading with fluo-4/AM in the presence of 1 mM La 3+ . Scale bars: 100 μm (a), 90 μm (b and c), 60 μm (d) and 70 μm (e).

    Article Snippet: Pollen tube fluorescence was developed and detected using the following steps: Exactly 50 μg of fluo-4/AM was dissolved in 91.2 μL of anhydrous dimethylsulfoxide (Dojindo) in a microcentrifuge tube.

    Techniques:

    A typical image showing changes in intracellular calcium concentrations from SPs. Balb/3T3 cells were seeded at 2×10 3 cells/well in culture medium. After incubation for 24 h, the cells were pretreated with the Fluo-4 AM mixture for 30 min. The intensity of intracellular green fluorescence was observed from 1 min before until 20 min after treatment with 300 µg/mL SP using a FluoView FV1000 confocal laser scanning microscope. The fluorescence of Fluo-4 was excited at 473 nm, and emission was detected at 516 nm. nSP70, closed circle (•); nSP300, closed triangle (▲); and mSP1000, closed square (■). The assay was performed in 3–5 independent experiments.

    Journal: Biochemistry and Biophysics Reports

    Article Title: Reduction of calcium flux from the extracellular region and endoplasmic reticulum by amorphous nano-silica particles owing to carboxy group addition on their surface

    doi: 10.1016/j.bbrep.2017.01.014

    Figure Lengend Snippet: A typical image showing changes in intracellular calcium concentrations from SPs. Balb/3T3 cells were seeded at 2×10 3 cells/well in culture medium. After incubation for 24 h, the cells were pretreated with the Fluo-4 AM mixture for 30 min. The intensity of intracellular green fluorescence was observed from 1 min before until 20 min after treatment with 300 µg/mL SP using a FluoView FV1000 confocal laser scanning microscope. The fluorescence of Fluo-4 was excited at 473 nm, and emission was detected at 516 nm. nSP70, closed circle (•); nSP300, closed triangle (▲); and mSP1000, closed square (■). The assay was performed in 3–5 independent experiments.

    Article Snippet: After incubation for 24 h, the cells were pretreated with Fluo-4 AM mixture in accordance with the attached protocol from Dojindo Laboratories.

    Techniques: Incubation, Fluorescence, Laser-Scanning Microscopy

    Calcium flux from ATP, nSP70, nSP70-COOH, nSP300, and mSP1000 in Balb/3T3. To determine the modulation of calcium flux by SPs in Balb/3T3, we performed Fluo-4 assays. The cells were pretreated with the Fluo-4 AM mixture for 30 min. The intensity of intracellular green fluorescence was observed from 1 min before until 20 min after treatment with ATP or SPs. The fluorescence of Fluo-4 is excited at 473 nm, and emission was detected at 516 nm. The assay was performed in 3–5 independent experiments; *P

    Journal: Biochemistry and Biophysics Reports

    Article Title: Reduction of calcium flux from the extracellular region and endoplasmic reticulum by amorphous nano-silica particles owing to carboxy group addition on their surface

    doi: 10.1016/j.bbrep.2017.01.014

    Figure Lengend Snippet: Calcium flux from ATP, nSP70, nSP70-COOH, nSP300, and mSP1000 in Balb/3T3. To determine the modulation of calcium flux by SPs in Balb/3T3, we performed Fluo-4 assays. The cells were pretreated with the Fluo-4 AM mixture for 30 min. The intensity of intracellular green fluorescence was observed from 1 min before until 20 min after treatment with ATP or SPs. The fluorescence of Fluo-4 is excited at 473 nm, and emission was detected at 516 nm. The assay was performed in 3–5 independent experiments; *P

    Article Snippet: After incubation for 24 h, the cells were pretreated with Fluo-4 AM mixture in accordance with the attached protocol from Dojindo Laboratories.

    Techniques: Fluorescence

    Determination of the underlying mechanism of promotion of calcium flux by nSP70. To determine the mechanism by which nSP70 promotes calcium flux, we analyzed the effects of a calcium channel or pump inhibitor, SKF96365 (SKF) or thapsigargin (Thap), on calcium flux. The cells were pretreated with each inhibitor for 2 h. They were then treated with Fluo-4 AM mixture for 30 min before the addition of nSP70. The intensity of intracellular green fluorescence was observed from 1 min before until 20 min after treatment with nSP70. Fluorescence of Fluo-4 was excited at 473 nm and emission was detected at 516 nm. The assay was performed in 3–5 independent experiments; *P

    Journal: Biochemistry and Biophysics Reports

    Article Title: Reduction of calcium flux from the extracellular region and endoplasmic reticulum by amorphous nano-silica particles owing to carboxy group addition on their surface

    doi: 10.1016/j.bbrep.2017.01.014

    Figure Lengend Snippet: Determination of the underlying mechanism of promotion of calcium flux by nSP70. To determine the mechanism by which nSP70 promotes calcium flux, we analyzed the effects of a calcium channel or pump inhibitor, SKF96365 (SKF) or thapsigargin (Thap), on calcium flux. The cells were pretreated with each inhibitor for 2 h. They were then treated with Fluo-4 AM mixture for 30 min before the addition of nSP70. The intensity of intracellular green fluorescence was observed from 1 min before until 20 min after treatment with nSP70. Fluorescence of Fluo-4 was excited at 473 nm and emission was detected at 516 nm. The assay was performed in 3–5 independent experiments; *P

    Article Snippet: After incubation for 24 h, the cells were pretreated with Fluo-4 AM mixture in accordance with the attached protocol from Dojindo Laboratories.

    Techniques: Fluorescence

    Cycling of Ca 2+ in CPCs in response to electrical stimulation is dependent on the LTCC, IP 3 R, and external Ca 2+ . (a) Representative Ca 2+ oscillations recorded from fluo-4/AM-loaded CPCs at 0.5 Hz electrical stimulation in 2 mM Ca 2+ Tyrode's solution, pretreated with nifedipine or 2-APB to inhibit the LTCC or IP 3 R, respectively, or in 0 Ca 2+ Tyrode's solution. Traces have been aligned in time based on when electrical stimulation was applied for comparison. Arrow indicates the start of stimulation. (b) Summary bar graph of Ca 2+ oscillation amplitudes under the various recording conditions in (a). Data represent mean ± SD. ∗ p

    Journal: Stem Cells International

    Article Title: Electrically Induced Calcium Handling in Cardiac Progenitor Cells

    doi: 10.1155/2016/8917380

    Figure Lengend Snippet: Cycling of Ca 2+ in CPCs in response to electrical stimulation is dependent on the LTCC, IP 3 R, and external Ca 2+ . (a) Representative Ca 2+ oscillations recorded from fluo-4/AM-loaded CPCs at 0.5 Hz electrical stimulation in 2 mM Ca 2+ Tyrode's solution, pretreated with nifedipine or 2-APB to inhibit the LTCC or IP 3 R, respectively, or in 0 Ca 2+ Tyrode's solution. Traces have been aligned in time based on when electrical stimulation was applied for comparison. Arrow indicates the start of stimulation. (b) Summary bar graph of Ca 2+ oscillation amplitudes under the various recording conditions in (a). Data represent mean ± SD. ∗ p

    Article Snippet: Electrical field stimulation was applied using an IonOptix MyoPacer Cell Stimulator (10 ms duration, 32 V; Westwood, MA, USA) and fluo-4 was excited at 488 nm and emission collected at 515 ± 15 nm using laser scanning confocal microscopy (FV1000, Olympus, Melville, NY, USA).

    Techniques:

    Nuclear Ca 2+ increases more than cytosolic Ca 2+ in response to electrical stimulation. Fluo-4/AM-loaded hcCPCs were stimulated at 0.5 Hz and the amplitudes of the nuclear and cytosolic Ca 2+ oscillations were analyzed. Representative traces of nuclear (red) and cytosolic (blue) Ca 2+ oscillations are shown in (a), and amplitudes are summarized in (b). Data represent mean ± SD. ∗ p

    Journal: Stem Cells International

    Article Title: Electrically Induced Calcium Handling in Cardiac Progenitor Cells

    doi: 10.1155/2016/8917380

    Figure Lengend Snippet: Nuclear Ca 2+ increases more than cytosolic Ca 2+ in response to electrical stimulation. Fluo-4/AM-loaded hcCPCs were stimulated at 0.5 Hz and the amplitudes of the nuclear and cytosolic Ca 2+ oscillations were analyzed. Representative traces of nuclear (red) and cytosolic (blue) Ca 2+ oscillations are shown in (a), and amplitudes are summarized in (b). Data represent mean ± SD. ∗ p

    Article Snippet: Electrical field stimulation was applied using an IonOptix MyoPacer Cell Stimulator (10 ms duration, 32 V; Westwood, MA, USA) and fluo-4 was excited at 488 nm and emission collected at 515 ± 15 nm using laser scanning confocal microscopy (FV1000, Olympus, Melville, NY, USA).

    Techniques:

    Ca 2+ oscillations in electrically stimulated CPCs show spatial heterogeneity and propagate in a wave-like fashion. ((a) and (b)) Representative 2D high-speed confocal image montage of the activation of a Ca 2+ oscillation in a CPC loaded with fluo-4/AM. ROIs (red: nuclear, blue: cytosolic) indicated in (a) are graphed in (b). ((c) and (d)) Measured from the red line shown in (c), the confocal line scan in (d) shows that initiation of Ca 2+ release at one edge of the cell propagates through the cell to the other side. (e) Representative image of a CPC stained with Di-8-ANEPPS.

    Journal: Stem Cells International

    Article Title: Electrically Induced Calcium Handling in Cardiac Progenitor Cells

    doi: 10.1155/2016/8917380

    Figure Lengend Snippet: Ca 2+ oscillations in electrically stimulated CPCs show spatial heterogeneity and propagate in a wave-like fashion. ((a) and (b)) Representative 2D high-speed confocal image montage of the activation of a Ca 2+ oscillation in a CPC loaded with fluo-4/AM. ROIs (red: nuclear, blue: cytosolic) indicated in (a) are graphed in (b). ((c) and (d)) Measured from the red line shown in (c), the confocal line scan in (d) shows that initiation of Ca 2+ release at one edge of the cell propagates through the cell to the other side. (e) Representative image of a CPC stained with Di-8-ANEPPS.

    Article Snippet: Electrical field stimulation was applied using an IonOptix MyoPacer Cell Stimulator (10 ms duration, 32 V; Westwood, MA, USA) and fluo-4 was excited at 488 nm and emission collected at 515 ± 15 nm using laser scanning confocal microscopy (FV1000, Olympus, Melville, NY, USA).

    Techniques: Activation Assay, Staining

    Whole-cell Ca 2+ oscillations in electrically stimulated CPCs. (a) 2D fluorescent image of child CPCs loaded with fluo-4/AM. Fluo-4/AM-loaded hcCPCs were electrically stimulated at 0.5 Hz (blue) or 1 Hz (orange). Representative recordings of Ca 2+ oscillations are shown in (b). The arrow indicates the start of electrical stimulation. The amplitude (Δ F / F 0 ) and frequency (oscillations/minute) of the oscillations are summarized in (c) and (d), respectively. (e) Representative recording of cytosolic Ca 2+ before, during, and after 0.5 Hz electrical stimulation. The bar above the trace shows the time during the recording when electrical stimulation was applied. Data in (e) represent mean ± SD. No significance was found by Student's t -test. n = 14 cells from 4 different pools of cells for all measurements.

    Journal: Stem Cells International

    Article Title: Electrically Induced Calcium Handling in Cardiac Progenitor Cells

    doi: 10.1155/2016/8917380

    Figure Lengend Snippet: Whole-cell Ca 2+ oscillations in electrically stimulated CPCs. (a) 2D fluorescent image of child CPCs loaded with fluo-4/AM. Fluo-4/AM-loaded hcCPCs were electrically stimulated at 0.5 Hz (blue) or 1 Hz (orange). Representative recordings of Ca 2+ oscillations are shown in (b). The arrow indicates the start of electrical stimulation. The amplitude (Δ F / F 0 ) and frequency (oscillations/minute) of the oscillations are summarized in (c) and (d), respectively. (e) Representative recording of cytosolic Ca 2+ before, during, and after 0.5 Hz electrical stimulation. The bar above the trace shows the time during the recording when electrical stimulation was applied. Data in (e) represent mean ± SD. No significance was found by Student's t -test. n = 14 cells from 4 different pools of cells for all measurements.

    Article Snippet: Electrical field stimulation was applied using an IonOptix MyoPacer Cell Stimulator (10 ms duration, 32 V; Westwood, MA, USA) and fluo-4 was excited at 488 nm and emission collected at 515 ± 15 nm using laser scanning confocal microscopy (FV1000, Olympus, Melville, NY, USA).

    Techniques:

    Δ F/F R responses measured from a midshipman swimbladder fiber that was injected with Fluo-4 and excited by a 67-Hz stimulus train that lasted 10 s. (A) The response during the first part of the recording, which included 0.05 s of baseline plus 0.45 s of stimulation. (B) The response during the last part of the recording (9–9.5 s after onset of stimulation).

    Journal: The Journal of General Physiology

    Article Title: Small Ca2+ releases enable hour-long high-frequency contractions in midshipman swimbladder muscle

    doi: 10.1085/jgp.201711760

    Figure Lengend Snippet: Δ F/F R responses measured from a midshipman swimbladder fiber that was injected with Fluo-4 and excited by a 67-Hz stimulus train that lasted 10 s. (A) The response during the first part of the recording, which included 0.05 s of baseline plus 0.45 s of stimulation. (B) The response during the last part of the recording (9–9.5 s after onset of stimulation).

    Article Snippet: For the latter reason, Mag-fluo-4 (Life Technologies) rather than furaptra was used as the primary Ca2+ indicator.

    Techniques: Injection

    Examples of force and Δ[Ca 2+ ] measurements from midshipman swimbladder fibers elicited by stimulus trains lasting many seconds. The labels on the left, middle, and right apply to both sides of the figure. (A, C, and E) Examples of force and Δ[Ca 2+ ] responses during the first ∼0.5 s of a long stimulus train at the indicated frequency. The trace in E reveals an obvious movement during the first part of the stimulus train. (B, D, and F) Responses from the same preparations in A, C, and E but during the last ∼0.5 s of stimulation. ΔT denotes the elapsed time between the recording of the traces in A, C, and E versus those in B, D, and F; the ΔT values were 4.3, 9.0, and 9.6 s, respectively. In D and F, the exact elevation of the Δ[Ca 2+ ] traces with respect to the baseline is not known, because data sampling was discontinued while records were stored to disc during the long stimulation period (at ΔT ∼3 and ∼6 s); thus, for data records after the first, the absolute fluorescence intensity of Mag-fluo-4, and hence the baseline elevation, is uncertain. The elevation of the Δ[Ca 2+ ] traces in D and F has been set, somewhat arbitrarily, at 0.5 µM (see Results). Experiment no. 052114.1 (C and D), 052114.3 (E and F).

    Journal: The Journal of General Physiology

    Article Title: Small Ca2+ releases enable hour-long high-frequency contractions in midshipman swimbladder muscle

    doi: 10.1085/jgp.201711760

    Figure Lengend Snippet: Examples of force and Δ[Ca 2+ ] measurements from midshipman swimbladder fibers elicited by stimulus trains lasting many seconds. The labels on the left, middle, and right apply to both sides of the figure. (A, C, and E) Examples of force and Δ[Ca 2+ ] responses during the first ∼0.5 s of a long stimulus train at the indicated frequency. The trace in E reveals an obvious movement during the first part of the stimulus train. (B, D, and F) Responses from the same preparations in A, C, and E but during the last ∼0.5 s of stimulation. ΔT denotes the elapsed time between the recording of the traces in A, C, and E versus those in B, D, and F; the ΔT values were 4.3, 9.0, and 9.6 s, respectively. In D and F, the exact elevation of the Δ[Ca 2+ ] traces with respect to the baseline is not known, because data sampling was discontinued while records were stored to disc during the long stimulation period (at ΔT ∼3 and ∼6 s); thus, for data records after the first, the absolute fluorescence intensity of Mag-fluo-4, and hence the baseline elevation, is uncertain. The elevation of the Δ[Ca 2+ ] traces in D and F has been set, somewhat arbitrarily, at 0.5 µM (see Results). Experiment no. 052114.1 (C and D), 052114.3 (E and F).

    Article Snippet: For the latter reason, Mag-fluo-4 (Life Technologies) rather than furaptra was used as the primary Ca2+ indicator.

    Techniques: Sampling, Fluorescence

    Examples of force responses and myoplasmic calcium transients from midshipman swimbladder fibers stimulated by APs. The labels on the far left and far right apply to both sides of the figure. (A) Examples of the twitch response from two different bundles of midshipman fibers excited by a one-shock field stimulus at zero time. (B) Force responses from the bundle corresponding to the upper trace in A in response to 67- and 100-Hz stimulation. (C) Mag-fluo-4 Δ F / F R and Δ[Ca 2+ ] signals during a twitch from one fiber on the surface of a swimbladder bundle; the signals were elicited by a pointwise (not field) stimulation. (D) Δ F / F R and Δ[Ca 2+ ] signals from the same fiber as in C when stimulated by 15 shocks at 100 Hz. (E and F) Responses like those in C and D from a different fiber and bundle; in F, the stimulus was 30 shocks at 67 Hz. The records in E and F reveal a slight a movement artifact. Experiment no. 051413.1 (C and D), 052114.1 (E and F).

    Journal: The Journal of General Physiology

    Article Title: Small Ca2+ releases enable hour-long high-frequency contractions in midshipman swimbladder muscle

    doi: 10.1085/jgp.201711760

    Figure Lengend Snippet: Examples of force responses and myoplasmic calcium transients from midshipman swimbladder fibers stimulated by APs. The labels on the far left and far right apply to both sides of the figure. (A) Examples of the twitch response from two different bundles of midshipman fibers excited by a one-shock field stimulus at zero time. (B) Force responses from the bundle corresponding to the upper trace in A in response to 67- and 100-Hz stimulation. (C) Mag-fluo-4 Δ F / F R and Δ[Ca 2+ ] signals during a twitch from one fiber on the surface of a swimbladder bundle; the signals were elicited by a pointwise (not field) stimulation. (D) Δ F / F R and Δ[Ca 2+ ] signals from the same fiber as in C when stimulated by 15 shocks at 100 Hz. (E and F) Responses like those in C and D from a different fiber and bundle; in F, the stimulus was 30 shocks at 67 Hz. The records in E and F reveal a slight a movement artifact. Experiment no. 051413.1 (C and D), 052114.1 (E and F).

    Article Snippet: For the latter reason, Mag-fluo-4 (Life Technologies) rather than furaptra was used as the primary Ca2+ indicator.

    Techniques: