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Fv1000mp

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The FV1000MP is a multiphoton confocal microscope system from Olympus. It is designed for high-resolution imaging and analysis of biological samples. The system combines a multiphoton excitation source and a confocal scanning unit to provide optical sectioning and high-contrast images.

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Lab products found in correlation

Sylgard, by Dow (4 mentions) Lipofectamine 2000, by Thermo Fisher Scientific (2 mentions) Dulbecco s modified eagle s medium, by Thermo Fisher Scientific (1 mentions) Cl 1000 uv crosslinker, by Analytik Jena (1 mentions) Xlpln25xwmp2, by Olympus (1 mentions) Bx61wi, by Olympus (1 mentions)

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FV1000MP microscope (6 citations)

7 protocols using fv1000mp

1

Visualizing G-quadruplex and 9CI in A549 cells

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A549 cells were cultured in Dulbecco's modified Eagle's medium (Gibco) with 1% glutamine and 10% fetal bovine serum (FBS) for 24 h at 37°C. 40 pmol annealed c-MYC Pu22, TTA and c-KIT1 G-quadruplex forming oligonucleotides labelled with 5′-Cy5 was transfected into cells by using lipofectamine 2000 (Thermo Fisher) and incubated at 37°C for 5 h. Then 5 μM 9CI was added and incubated for another 2 h. The digital images were recorded using a confocal laser scanning microscopy (Olympus FV1000-MP). The location of 9CI in the cells was investigated by the fluorescence signal collected between 425–470 nm/500–545 nm at λex = 405 nm, and the location of G-quadruplex was checked by fluorescence signal collected between 655 and 755 nm with the excitation at 635 nm.
Zhai Q., Gao C., Ding J., Zhang Y., Islam B., Lan W., Hou H., Deng H., Li J., Hu Z., Mohamed H.I., Xu S., Cao C., Haider S.M, & Wei D. (2019). Selective recognition of c-MYC Pu22 G-quadruplex by a fluorescent probe. Nucleic Acids Research, 47(5), 2190-2204.
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2

Calcium Imaging of C4da Neuron Terminals

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Calcium imaging of C4da axon terminals was performed as previously described (Figure 4—figure supplement 1; Hu et al., 2017 (link)). Briefly, staged third instar larvae (96 ± 3 hr AEL) were pinned on a Sylgard (Dow Corning) plate and partially dissected in physiological saline to expose the ventral nerve cord (VNC). C4da neuron axon terminals expressing GCaMP6m were live-imaged by confocal microscopy with a 40 × water objective (Olympus FV1000MP) with 3x zoom to image at least four segments ensuring the calcium response could be detected. Activation of sensory neurons was achieved by providing a mechanonociceptive cue using a micromanipulator-mounted von Frey filament (45 mN) for stimulation of midabdominal segments (A3–A5). The most robust responses to local von Frey filament stimulation are restricted to a single VNC hemisegment corresponding to the stimulation site on the body wall although the adjacent segment(s) could also be slightly activated. The transient dip in fluorescence intensity (Figure 4B) is due to the larval brain moving out of focus briefly during and after mechanical stimulation. Baseline (F0) and relative maximum intensity change (ΔFmax) of GCaMP6m fluorescence were analysed.
Dannhäuser S., Lux T.J., Hu C., Selcho M., Chen J.T., Ehmann N., Sachidanandan D., Stopp S., Pauls D., Pawlak M., Langenhan T., Soba P., Rittner H.L, & Kittel R.J. (2020). Antinociceptive modulation by the adhesion GPCR CIRL promotes mechanosensory signal discrimination. eLife, 9, e56738.
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3

Photochemical Activation of Peptides

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Light activation of peptides for biochemical studies was performed using a CL-1000 UV Crosslinker (UVP, 8 mW/cm2). Power density was measured by an OAI 306 UV power meter at 365 nm. Typical exposure time for these studies was 10–30 minutes. For activation in cell and animal studies, Lumen Dynamics UV system with 365 nm fiber light guide was used (OmniCure 1000, 200 mW/cm2). For in vivo activation at the tumor site, mice were anesthetized and the light was guided through an optical cable and placed approximately 3 cm from the flank tumor. Each flank tumor was exposed for 30 seconds.
Two-photon unveiling was performed at the KI Microscopy Core with a multiphoton microscope (Olympus FV-1000MP) operating at 690 nm with a Spectra-Physics Deepsea Tia-sapphire laswer at power 1 W using a 25× objective with 1.05 NA. Samples were placed in glass bottom 384 well plates. Images were captured at 840 nm.
Dudani J.S., Jain P.K., Kwong G.A., Stevens K.R, & Bhatia S.N. (2015). Photoactivated Spatiotemporally-Responsive Nanosensors of In Vivo Protease Activity. ACS nano, 9(12), 11708-11717.
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4

Two-Photon Imaging of Dendritic Morphology

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Two-photon imaging was performed using an upright microscope (models BX61WI and FV1000-MP, Olympus) equipped with a resonant scanner and a GaAsP detector with a 25× (1.05 numerical aperture; catalog #XLPLN25XWMP2, Olympus) water-immersion lens. A Ti-sapphire laser (Mai-Tai-DS-HP, SpectraPhysics) was tuned to 950 nm wavelength for the excitation of the GFP. The average excitation power was maintained at <40 mW under the objective. The power was selected to avoid saturating the fluorescence of dendrites. Mice were anesthetized with a 0.8–1.5% isoflurane–oxygen mixture (Univentor 400 anesthesia unit, Univentor), and the body temperature was maintained at 37°C using a heating pad. During the image acquisition, mice were restrained using a head plate. Image stacks were obtained from the apical dendritic tufts of layer V pyramidal in the layer I (20–70 μm from the pial surface), and, typically, 20–40 frames (512 × 512 pixels; 0.124 μm/pixel; 17 ms/frame) per stack were obtained at an interval of 0.4 μm along the z-axis. Each dendrite was traced to the soma to confirm its layer of origin.
Ishii K., Nagaoka A., Kishida Y., Okazaki H., Yagishita S., Ucar H., Takahashi N., Saito N, & Kasai H. (2018). In Vivo Volume Dynamics of Dendritic Spines in the Neocortex of Wild-Type and Fmr1 KO Mice. eNeuro, 5(5), ENEURO.0282-18.2018.
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5

Live Imaging of Sensory Neuron Activation

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Staged 3rd instar larvae (96h AEL±3h AEL) were pinned on a Sylgard (Dow Corning) plate and partially dissected in physiological saline21 (link) to expose the VNC. A08n or DP-ilp7 neuron somata or C4da neuron axon terminals expressing GCaMP6m were live imaged by confocal microscopy with a 40× water objective (Olympus FV1000MP). Activation of sensory neurons was achieved either by class specific expression of CsChrimson (625 nm/5 s, 0.2 mW/mm2) or providing a mechano-nociceptive cue using a micromanipulator mounted von Frey filament (45 mN) and stimulation of mid-abdominal segments (a3–5). Baseline (F0) and relative maximum intensity change (Fmax) of GCaMP6m fluorescence was analyzed. Fmax/F0 values were represented as box-plots with whiskers, with the centerline representing median values, upper and lower edges of the boxes representing the 25th and 75th percentiles of the sample data, respectively. The short line within the box represents the mean of the data. Upper and lower whiskers represent the 5th and 95th percentile of the sample data, respectively. Statistical significance was analyzed using Mann-Whitney U-test with Bonferroni correction for multiple comparisons.
Hu C., Petersen M., Hoyer N., Spitzweck B., Tenedini F., Wang D., Gruschka A., Burchardt L.S., Szpotowicz E., Schweizer M., Guntur A.R., Yang C.H, & Soba P. (2017). Modality-specific sensory integration and neuropeptide-mediated feedback facilitate mechano-nociceptive behavior in Drosophila. Nature neuroscience, 20(8), 1085-1095.
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6

Live Imaging of Sensory Neuron Activation

Cited 1 time
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Staged 3rd instar larvae (96h AEL±3h AEL) were pinned on a Sylgard (Dow Corning) plate and partially dissected in physiological saline21 (link) to expose the VNC. A08n or DP-ilp7 neuron somata or C4da neuron axon terminals expressing GCaMP6m were live imaged by confocal microscopy with a 40× water objective (Olympus FV1000MP). Activation of sensory neurons was achieved either by class specific expression of CsChrimson (625 nm/5 s, 0.2 mW/mm2) or providing a mechano-nociceptive cue using a micromanipulator mounted von Frey filament (45 mN) and stimulation of mid-abdominal segments (a3–5). Baseline (F0) and relative maximum intensity change (Fmax) of GCaMP6m fluorescence was analyzed. Fmax/F0 values were represented as box-plots with whiskers, with the centerline representing median values, upper and lower edges of the boxes representing the 25th and 75th percentiles of the sample data, respectively. The short line within the box represents the mean of the data. Upper and lower whiskers represent the 5th and 95th percentile of the sample data, respectively. Statistical significance was analyzed using Mann-Whitney U-test with Bonferroni correction for multiple comparisons.
Hu C., Petersen M., Hoyer N., Spitzweck B., Tenedini F., Wang D., Gruschka A., Burchardt L.S., Szpotowicz E., Schweizer M., Guntur A.R., Yang C.H, & Soba P. (2017). Modality-specific sensory integration and neuropeptide-mediated feedback facilitate mechano-nociceptive behavior in Drosophila. Nature neuroscience, 20(8), 1085-1095.
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7

Calcium Imaging of C4da Neuron Terminals

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Calcium imaging of C4da axon terminals was performed as previously described (Hu et al., 2017) . Briefly, staged third instar larvae (96 ± 3 h AEL) were pinned on a Sylgard (Dow Corning) plate and partially dissected in physiological saline to expose the ventral nerve cord (VNC). C4da neuron axon terminals expressing GCaMP6m were live-imaged by confocal microscopy with a 40× water objective (Olympus FV1000MP) with 3x zoom to image at least 4 segments ensuring the calcium response could be detected. Activation of sensory neurons was achieved by providing a mechanonociceptive cue using a micromanipulator-mounted von Frey filament (45 mN) for stimulation of midabdominal segments (A3-A5). The most robust responses to local von Frey filament stimulation are restricted to a single VNC hemisegment corresponding to the stimulation site on the body wall although the adjacent segment(s) could also be slightly activated. Baseline (F0) and relative maximum intensity change (ΔFmax) of GCaMP6m fluorescence were analysed.
Dannhäuser S., Lux T.J., Hu C., Selcho M., Chen J.T., Ehmann N., Sachidanandan D., Pawlak M., Langenhan T., Soba P., Rittner H.L., & Kittel R.J. (2020). Antinociceptive modulation by the adhesion-GPCR CIRL promotes mechanosensory signal discrimination.
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