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Sp5 2 sted cw

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The SP5 II STED-CW is a laser scanning confocal microscope system designed for advanced imaging techniques, including Stimulated Emission Depletion (STED) microscopy. The system features a continuous wave (CW) STED laser for high-resolution imaging. The core function of the SP5 II STED-CW is to provide researchers with a versatile and powerful tool for high-resolution, nanoscale imaging applications.

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Lab products found in correlation

Live dead cell viability assay kit, by Thermo Fisher Scientific (2 mentions) Hcx pl apo, by Leica (2 mentions) Lysotracker green dnd 26, by Thermo Fisher Scientific (1 mentions) 7 aad, by BioLegend (1 mentions) Fluorodish, by World Precision Instruments (1 mentions)

Close spelling variants of this product

SP5-II-STED-CW confocal microscope SP5 II STED-CW Superresolution Laser Scanning Confocal microscope TCS SP5 II CW STED STED CW SP5 II

8 protocols using sp5 2 sted cw

1

Imaging Purified Recombinant Proteins

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Purified recombinant proteins were imaged using either 63x or 100x oil immersion objectives, on a Leica SP5 II STED-CW super-resolution laser scanning confocal microscope with Gallium arsenide phosphide (GaAsP) / Photon Multiplier Tube (PMT) hybrid detectors and LAS_AF Leica proprietary software. For in vitro studies, purified recombinant protein was subjected to brief heat shock (when applicable) using an Eppendorf ThermoMixer F1.5 heat block, deposited on a slide and immediately imaged. For imaging experiments performed between pH 5.7 and pH 6.4, purified protein samples were buffered in 20mM HEPES, 150mM KCl. For imaging experiments performed between pH 5.0 and 5.6, purified protein samples were buffered in 20mM sodium acetate, 150mM KCl.
Riback J.A., Katanski C.D., Kear-Scott J.L., Pilipenko E.V., Rojek A.E., Sosnick T.R, & Drummond D.A. (2017). Stress-triggered phase separation is an adaptive, evolutionarily tuned response. Cell, 168(6), 1028-1040.e19.
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2

Immunostaining of Skeletal Muscle Fibers

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The flexor digitorum brevis (FDB) muscle bundle was dissected and placed in 1ml of DMEM containing BSA plus collagenase solution pre-warmed to 37°C in a 12-well plate. After 2 hours, fibers were triturated in Ringers solution. Fibers were fixed on coverslips with 4% paraformaldehyde blocked in 1X phosphate-buffered saline (PBS) containing 10% fetal bovine serum and triton, and then immunostained at 1:100 with anti-DHPR (ma3-920, Thermo Scientific, Rockford, IL) and at 1:100 with anti-BIN1(sc-30099, Santa Cruz, Dallas, TX). The anti-EHD1 rabbit antibody was previously described [10 (link)]. Goat anti-rabbit conjugated to Alexa 488 and goat anti-mouse 594 were used at 1:2000. Slides were mounted with Vectashield with DAPI. Images were captured using a Leica SP5 II STED-CW super resolution laser scanning confocal microscope in standard mode.
Demonbreun A.R., Swanson K.E., Rossi A.E., Deveaux H.K., Earley J.U., Allen M.V., Arya P., Bhattacharyya S., Band H., Pytel P, & McNally E.M. (2015). Eps 15 Homology Domain (EHD)-1 Remodels Transverse Tubules in Skeletal Muscle. PLoS ONE, 10(9), e0136679.
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3

Intracellular Localization of TAT-Au NP-Dox

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5×104 U87 glioma cells were seeded into glass-bottom culture dishes. TAT-Au NP-Dox, Au NP-Dox and Dox in 10% FBS DMEM was added to the dishes at the Dox concentration of 1 μM, respectively. The treated cells were monitored at 4 and 24 hours post-incubation via confocal microscopy. To determine the location of intracellular lysosomes, the treated cells were washed 3 times with PBS and incubated with LysoTracker Green DND-26 (Invitrogen, USA) at 7.5×10-8 M for 30 minutes. Live cell images were taken by a Leica SP5 II STED-CW super-resolution laser scanning confocal microscope.
Cheng Y., Dai Q., Morshed R., Fan X., Wegscheid M.L., Wainwright D.A., Han Y., Zhang L., Auffinger B., Tobias A.L., Rincón E., Thaci B., Ahmed A.U., Warnke P., He C, & Lesniak M.S. (2014). Blood-Brain Barrier Permeable Gold Nanoparticles: An Efficient Delivery Platform for Enhanced Malignant Glioma Therapy and Imaging. Small (Weinheim an der Bergstrasse, Germany), 10(24), 5137-5150.
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4

Investigating SD Particle Uptake and Release

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2x104 HB1.F3.CD cells or U87 cells were seeded on top of microscope glass coverslips. SD particles (106 SD/mL) were added to the cells. After 24 hours incubation, cells were washed 3 times with 10% FBS DMEM. To assess the permeability, cells were incubated with 7-AAD (Biolegend, USA) in DMEM and then received the MF treatment. Images were taken by a Leica SP5 II STED-CW super-resolution laser scanning confocal microscope. To study the SD release, 2x104 SD-loaded HB1.F3.CD cells were co-cultured with U87-GFP-Luc on microscope glass coverslips and treated under MF for 30 min. Confocal images were taken 24 hours post treatment to identify SD particles in glioma cells.
Muroski M.E., Morshed R.A., Cheng Y., Vemulkar T., Mansell R., Han Y., Zhang L., Aboody K.S., Cowburn R.P, & Lesniak M.S. (2016). Controlled Payload Release by Magnetic Field Triggered Neural Stem Cell Destruction for Malignant Glioma Treatment. PLoS ONE, 11(1), e0145129.
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5

Spinning Disk Confocal Microscopy Imaging

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Images were taken by a Marianas Yokogawa-type spinning disk (inverted confocal microscope). The following lasers were used: (1) green: λexc = 488 nm and green filter; (2) red: λexc = 565 nm and red filter; (3) transferrin: λexc = 640 nm and far-red filter; and (4) FRET channel: λexc = 488 nm and red filter. Super-resolution images were taken on a Leica SP5 II STED-CW super-resolution laser scanning confocal microscope. All imaging was performed at the Integrated Light Microscope Core Facility at the University of Chicago. Images were analyzed by ImageJ software.
Acar H., Samaeekia R., Schnorenberg M.R., Sasmal D.K., Huang J., Tirrell M.V, & LaBelle J.L. (2017). Cathepsin-Mediated Cleavage of Peptides from Peptide Amphiphiles Leads to Enhanced Intracellular Peptide Accumulation. Bioconjugate chemistry, 28(9), 2316-2326.
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6

Optogenetic Stimulation of DRG Neurons

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DRG neurons were cultured onto glass bottom petri dishes. PIN-SiNWs were sonicated into culture medium, drop casted onto the cells, and left to be co-cultured with the cells for 24 hours. For experiments without light stimulation, cells were stained with a LIVE/DEAD cell viability assay kit (ThermoFisher Scientific) and the numbers of live cells in culture with and without nanowires were counted. For experiments with light stimulation, cells were stimulated via the 592 nm depletion laser on an SP5 laser scanning confocal microscope (Leica, USA, SP5 II STED-CW) under a 40× objective (Leica, USA, HCX PL APO) at various frequencies for various durations at a total energy density of 0.31 μJ/μm2 (link) for each pulse. After stimulation, cells were stained with a LIVE/DEAD cell viability assay kit (ThermoFisher Scientific) and the numbers of live stimulated neurons, neurons neighboring the stimulated neurons, and unstimulated neurons were counted.
Parameswaran R., Carvalho-de-Souza J.L., Jiang Y., Burke M.J., Zimmerman J.F., Koehler K., Phillips A., Yi J., Adams E.J., Bezanilla F, & Tian B. (2018). Photoelectrochemical modulation of neuronal activity with free-standing coaxial silicon nanowires. Nature nanotechnology, 13(3), 260-266.
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7

Optogenetic Stimulation of DRG Neurons

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DRG neurons were cultured onto glass bottom petri dishes. PIN-SiNWs were sonicated into culture medium, drop casted onto the cells, and left to be co-cultured with the cells for 24 hours. For experiments without light stimulation, cells were stained with a LIVE/DEAD cell viability assay kit (ThermoFisher Scientific) and the numbers of live cells in culture with and without nanowires were counted. For experiments with light stimulation, cells were stimulated via the 592 nm depletion laser on an SP5 laser scanning confocal microscope (Leica, USA, SP5 II STED-CW) under a 40× objective (Leica, USA, HCX PL APO) at various frequencies for various durations at a total energy density of 0.31 μJ/μm2 (link) for each pulse. After stimulation, cells were stained with a LIVE/DEAD cell viability assay kit (ThermoFisher Scientific) and the numbers of live stimulated neurons, neurons neighboring the stimulated neurons, and unstimulated neurons were counted.
Parameswaran R., Carvalho-de-Souza J.L., Jiang Y., Burke M.J., Zimmerman J.F., Koehler K., Phillips A., Yi J., Adams E.J., Bezanilla F, & Tian B. (2018). Photoelectrochemical modulation of neuronal activity with free-standing coaxial silicon nanowires. Nature nanotechnology, 13(3), 260-266.
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8

Laser-Induced Nanowire-Bacterial Interactions

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The overnight LB-cultured E. coli or B. subtilis was diluted to OD600 = 1 and mixed with mesostructured intrinsic nanowires. The solution was added on a FluoroDish (World Precision Instruments) and covered with a coverslip. The single-cell manipulation was performed using a confocal laser scanning microscope (Leica, SP5 II STED-CW). In a typical experiment, a laser pulse (1 ms, 592 nm) was delivered to a nanowire/bacterial cell of interest in the middle of an imaging time series.
Gao X., Jiang Y., Lin Y., Kim K.H., Fang Y., Yi J., Meng L., Lee H.C., Lu Z., Leddy O., Zhang R., Tu Q., Feng W., Nair V., Griffin P.J., Shi F., Shekhawat G.S., Dinner A.R., Park H.G, & Tian B. (2020). Structured silicon for revealing transient and integrated signal transductions in microbial systems. Science Advances, 6(7), eaay2760.
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