HeLa cells were first seeded on coverslips in petri dishes with DMEM culture medium at 37 °C for 24 h. ER-GFP (C10590, Invitrogen) and Actin-RFP (C10583, Invitrogen) plasmids were transfected into cells for 48 h following the protocol from Invitrogen. 500 μM oleic acid (O1383 SIGMA) coupled with BSA in DMEM culture medium was then added to cells for 7 h to induce the formation of lipid droplets. Before imaging, cells were first incubated with 6 μM SYTO60 (S11342, Invitrogen), 120 nM LysoTracker Deep Red (L12492, Invitrogen) and 400 nM Rhodamine 800 (83701 SIGMA) in HBSS simultaneously for 30 min at 37 °C, followed by staining with NucBlue® Live ReadyProbes® Reagent (R37605, Invitrogen) in HBSS for 20 min at 37 °C. Then cells were incubated with ATTO 740-conjugated WGA in HBSS for 30 min at 37 °C, followed by staining with LipidTOX Deep Red (H34477, Invitrogen) in HBSS with a dilution of 1:20 for 30 min at room temperature before imaging.
Rhodamine 800
Manufactured by Merck Group
Sourced in Sao Tome and Principe, United States
Rhodamine 800 is a fluorescent dye commonly used in laboratory settings. It is a synthetic organic compound with a distinctive red-orange color. Rhodamine 800 exhibits strong fluorescence properties, making it a valuable tool for various applications in scientific research and analysis.
Automatically generated - may contain errors
Lab products found in correlation
Syto60, by Thermo Fisher Scientific (2 mentions)
Oleic acid, by Merck Group (2 mentions)
Nucblue live readyprobes reagent, by Thermo Fisher Scientific (2 mentions)
Lysotracker deep red, by Thermo Fisher Scientific (2 mentions)
Lipidtox deep red, by Thermo Fisher Scientific (2 mentions)
C10590, by Thermo Fisher Scientific (2 mentions)
Actin rfp, by Thermo Fisher Scientific (2 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Mitotracker deep red fm, by Thermo Fisher Scientific (1 mentions)
Mitoview 633, by Biotium (1 mentions)
Trizol, by Thermo Fisher Scientific (1 mentions)
Rhodamine 123, by Merck Group (1 mentions)
Claudin 5, by Merck Group (1 mentions)
β actin antibody, by Merck Group (1 mentions)
β catenin, by Merck Group (1 mentions)
P glycoprotein, by Abcam (1 mentions)
Irdye 800 cw peg, by LI COR (1 mentions)
Ebm 2 media, by Lonza (1 mentions)
Claudin 1, by Abcam (1 mentions)
Hela cells atcc ccl 2, by American Type Culture Collection (1 mentions)
8 protocols using rhodamine 800
1
Multicolor Labeling for Lipid Droplet Imaging
2
Mitochondrial Labeling with Photosensitizers
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Mitotracker Deep Red FM (Life Technologies, #M22426), Mitoview 633 (Biotium, #70055), Rhodamine 800 (Sigma, #83701), and TMRE (Life Technologies, #T-669) were used without further purification. Photosensitizer stock solutions were prepared in DMSO (Sigma, #D2650). Absorption spectra of the photosensitizers were determined on a UV-Vis spectrophotometer (Beckman Coulter, #DU-800) in PBS buffer (pH 7.4) at room temperature within a quartz cuvette (Nova Biotech, #QS-467). Condition for labeling HeLa cell mitochondria with photosensitizers: 100 nM in complete medium at 37 oC for 30 min (followed by 3X PBS wash).
3
Modeling Blood-Brain Barrier Interactions
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hCMEC/D3 were obtained from Dr. Peter Couraud59 (link) INSERM, France. WntC59 was purchased from AdooQ Biosciences (Irvine, CA). Human Recombinant Wn3a, and GF120918 (GF) were purchased from R&D Systems (Minneapolis, MN). Rhodamine 800 and Rhodamine 123 were purchased from Sigma (St. Louis, MO). IRDye 800 CW PEG were purchased from LICOR (Lincoln, NB). EBM-2 media was purchased from Lonza (Walkersville, MD). The E-plates for RTCA were obtained from ACEA Biosciences (San Diego, CA). Transwell and tissue culture plates were purchased from Corning (Tewksbury, MA). Trizol and tetramethylrhodamine BSA was purchased from Life Technologies (CA, USA). P-glycoprotein and Claudin-1 antibodies were purchased from Abcam (Cambridge, MA). β-catenin, claudin-5 and β-actin antibodies were purchased from Sigma (St. Louis, MO). Human fetal brain total RNA was purchased from Takara Bio USA, Inc. (Madison, WI). Primers were obtained from Invitrogen (CA, USA) and primer sequence information is provided in Table 1 Supplementary information.
4
HeLa Cell Staining with Rhodamine 800
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HeLa (ATCC CCL-2) cells were purchased from ATCC. Rhodamine 800 (Sigma 83701) and all solvents including hexanes, tetrahydrofuran (THF), dimethylformamide (DMF), chloroform, dichloromethane (DCM), acetone, and ethyl acetate were obtained from Sigma-Aldrich.
5
Multicolor Labeling for Lipid Droplet Imaging
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HeLa cells were first seeded on coverslips in petri dishes with DMEM culture medium at 37 °C for 24 h. ER-GFP (C10590, Invitrogen) and Actin-RFP (C10583, Invitrogen) plasmids were transfected into cells for 48 h following the protocol from Invitrogen. 500 μM oleic acid (O1383 SIGMA) coupled with BSA in DMEM culture medium was then added to cells for 7 h to induce the formation of lipid droplets. Before imaging, cells were first incubated with 6 μM SYTO60 (S11342, Invitrogen), 120 nM LysoTracker Deep Red (L12492, Invitrogen) and 400 nM Rhodamine 800 (83701 SIGMA) in HBSS simultaneously for 30 min at 37 °C, followed by staining with NucBlue® Live ReadyProbes® Reagent (R37605, Invitrogen) in HBSS for 20 min at 37 °C. Then cells were incubated with ATTO 740-conjugated WGA in HBSS for 30 min at 37 °C, followed by staining with LipidTOX Deep Red (H34477, Invitrogen) in HBSS with a dilution of 1:20 for 30 min at room temperature before imaging.
6
Rhodamine-based Hepatocyte Metabolism Assay
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The materials were from following sources: Rhodamine B, Rhodamine 800, collagenase type II, forskolin, and D-glucose from Sigma-Aldrich (MO, USA); William’s E medium, fetal bovine serum and Hank’s Balanced Salt Solution from Thermo Fisher Scientific Co. Ltd. (Waltham, MA, USA); matrigel was from BD Biosciences Company (New York, USA); coverslips were from Glaswarenfabrik Karl Hecht GmbH & Co KG (Rhön, Germany); PGE2, L161, 982, L798, 106, AH6809 and 3,7-Dihydro-1-methyl-3-(2-methylpropyl)-1H-purine-2,6-dione(IBMX) from Cayman (Michigan, USA); Percoll from GE Healthcare (Freiburg, Germany); CCCP from ABCAM (Cambridge, MA, USA); ATP assay kit from Beyotime (Shanghai, China); cAMP assay kit from R&D systems (Minnesota, USA); Bradford reagent for protein concentration analysis from BIO-RAD (Berkeley, California, USA). Rhodamine B-Me was synthesized as recently reported13 (link).
7
Somatostatin Receptor Targeting for Osteosarcoma Imaging
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Human osteosarcoma cells (U2OS), either transfected with SSTR2 [20 (link)] or wild-type (without SSTR2 expression) were cultured in DMEM (Gibco Life Technologies, Waltham, MA, USA) with 10% fetal calf serum and 1% penicillin. The cells were seeded in 24 well plates (105 cells per well) and grown until confluent. The cells were kept alive or killed with 50 µL EtOH (70%) and incubated with 800CW-TATE (10 nM, piCHEM, Grambach, Austria) in culturing medium for 30 min. A blocking experiment was included by co-incubation of 800CW-TATE (10 nM) with DOTA-TATE (10 µM, Bachem, Bubendorf, Switzerland). Control experiments were included by incubation of the cells with a NIR-fluorescent dye that does not bear a cyanine motive: Rhodamine 800 (10 nM, Sigma-Aldrich). Additionally, wells without seeded cells were exposed to PBS or EtOH and then incubated with 800CW-TATE, to investigate non-specific binding to plastic. After gentle washing with PBS, NIR-fluorescence imaging of the whole plate was performed on an Odyssey flatbed scanner (Li-Cor, Lincoln, USA). 800CW-TATE binding was based on fluorescent signal ± SD and each experiment was performed in n = 12.
8
Synthesis and Characterization of Luminescent Polymers
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All organic solvents were purchased from Wako Pure Chemical Industories (Tokyo, Japan). Poly(methyl methacrylate) (PMMA) (M w : 996 000), polystyrene(PS) (M w : 280 000), Rhodamine 800 were purchased from Sigma-Aldrich (St. Louis, MO). Poly(vinyl alcohol) (PVA) (M w : 22 000) was purchased from Kanto Chemical Industry (Tokyo, Japan). Eu-tris (dinaphthoylmethane)-bistrioctylphosphine oxide (EuDT) was synthesized according to the previous literature. ( 27) Poly(ethylene terephtalate) (PET, Lumirror T60, thickness: 25 μm) was purchased from Toray Industries Inc. (Tokyo, Japan).
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