Tcs sp2 aobs

Manufactured by Leica

Sourced in Germany, United Kingdom, Japan, United States

The TCS SP2 AOBS is a confocal laser scanning microscope produced by Leica. It is designed for high-resolution fluorescence imaging and features an acousto-optical beam splitter (AOBS) for flexible control of laser lines. The TCS SP2 AOBS provides researchers with a powerful tool for advanced microscopy applications.

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Lab products found in correlation

4 6 diamidino 2 phenylindole (dapi), by Merck Group (6 mentions) 4 6 diamidino 2 phenylindole (dapi), by Vector Laboratories (5 mentions) 4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (4 mentions) Alexa fluor 488 conjugated goat anti mouse, by Thermo Fisher Scientific (4 mentions) N cadherin, by BD (4 mentions) 4 6 diamidino 2 phenylindole dihydrochloride dapi, by Merck Group (3 mentions) Triton x 100, by Merck Group (3 mentions) Alexa fluor 568, by Thermo Fisher Scientific (3 mentions) Imagexpress micro, by Molecular Devices (2 mentions) Alexa 568, by Thermo Fisher Scientific (2 mentions) Prolong diamond antifade mountant containing 4 6 diamidino 2 phenylindole dapi, by Thermo Fisher Scientific (2 mentions) Hoechst 33342, by Merck Group (2 mentions) Facscalibur, by BD (2 mentions) Alexa 546 conjugated anti rabbit antiserum, by Thermo Fisher Scientific (2 mentions) Hcx pl apo cs 40 1.25 oil uv objective, by Leica (2 mentions) Triton, by Merck Group (2 mentions) Paraformaldehyde (pfa), by Merck Group (2 mentions) Sc 14365, by Santa Cruz Biotechnology (2 mentions) Cp290a, by Biocare Medical (2 mentions) Nb00 134, by Novus Biologicals (2 mentions)

Close spelling variants of this product

TCS SP2 (888 citations) TCS SP2 AOBS confocal microscope (95 citations) SP2 AOBS (45 citations) TCS-SP2 AOBS Confocal Laser Scanning Microscope (44 citations) TCS SP2 AOBS confocal system (14 citations) TCS SP2 AOBS MP (14 citations) TCS SP2 AOBS confocal laser microscope (13 citations) TCS SP2 AOBS microscope (9 citations) TCS SP2 AOBS spectral confocal microscope (7 citations) TCS SP2 AOBS system (6 citations)

189 protocols using tcs sp2 aobs

Immunofluorescence Microscopy Protocol

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Cells were fixed in 4% formaldehyde in PBS for 30 minutes at RT, washed with PBS followed by 15 minutes of 0.2% Triton X-100 permeabilization on ice. 3% BSA in 1xPBS was used for blocking 30 minutes at RT. Primary antibodies were added overnight at 4°C (Hoechst and fluorescent secondary antibodies were added post washing for 1 to 2 hrs at RT). Images were taken either with a 20X air objective, an automated inverted epifluorescence microscope (ImageXpressMicro, Molecular Devices), with a 40X or 100X oil objective on a confocal microscope (Leica TCS SP2 AOBS or 3i imaging system). See section on Fixed/live cell imaging for more detailed description. See Supplementary Table 5 for antibody information.

Gan L., Seki A., Shen K., Iyer H., Han K., Hayer A., Wollman R., Ge X., Lin J., Dey G., Talbot W.S, & Meyer T. (2019). The lysosomal GPCR-like protein GPR137B regulates Rag and mTORC1 localization and activity. Nature cell biology, 21(5), 614-626.

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Immunofluorescence Staining of Cells

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Cells were grown to 50% confluence on cover slip slides (12-545-80, Fisherbrand* Cover Glasses) overnight and fixed first with 4% formaldehyde in PBS (phosphate Buffered Saline) for 15 min at room temperature. Fixed cells were blocked with blocking buffer composed of 1xPBS with 5% goat serum (Cell Signaling Technology 5425, Danvers, MA) and 0.3% Triton X-100 for 60 minutes at room temperature. Cells were then probed with an anti-FLAG (Sigma F1804, Saint Louis, MO) sera at dilution of 1:200 and β-catenin (D10A8) XP Rabbit antibody (Cell Signaling Technology 8480, Danvers, MA) at dilution of 1:100 for 2 hours at room temperature, washed with PBS buffer three times of 5 min each, then incubated with secondary Alexa Fluor® 594 Goat Anti-Mouse IgG, highly cross-adsorbed antibody (Life Technologies A31624, Carlsbad, CA) and Alexa Fluor® 488 Goat Anti-Rabbit IgG (H+L), highly cross-adsorbed antibody (Life Technologies A11034, Carlsbad, CA) at a dilution of 1:1000 for 30 min at room temperature in dark. Nuclei were stained with DAPI (Cell Signaling Technology 8961, Danvers, MA). Next, the Slides were rinsed with PBS and Prolong Gold Anti-Fade Reagent (Cell Signaling Technology 9071, Danvers, MA) was applied. Slides were then analysed and images were taken on Leica TCS SP2 AOBS filter-free UV/spectral confocal laser scanner on an inverted DM IRE2 microscope.

Song J., Du Z., Ravasz M., Dong B., Wang Z, & Ewing R.M. (2015). A protein interaction between β-catenin and Dnmt1 regulates Wnt Signaling and DNA methylation in colorectal cancer cells. Molecular cancer research : MCR, 13(6), 969-981.

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Confocal Laser Scanning Microscopy Imaging

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Images were obtained using an inverted confocal laser scanning microscope
(Leica TCS SP2-AOBS; Mannheim, Germany) equipped with an UV-diode
(405 nm), argon (458, 476, 488, 514 nm), DPSS (561 nm)
and 2 He–Ne laser (594, 633 nm). Pictures were taken with a 63x
Plan-Apo, 1.4NA oil immersion objective.

Hassouna I., Ott C., Wüstefeld L., Offen N., Neher R.A., Mitkovski M., Winkler D., Sperling S., Fries L., Goebbels S., Vreja I.C., Hagemeyer N., Dittrich M., Rossetti M.F., Kröhnert K., Hannke K., Boretius S., Zeug A., Höschen C., Dandekar T., Dere E., Neher E., Rizzoli S.O., Nave K.A., Sirén A.L, & Ehrenreich H. (2016). Revisiting adult neurogenesis and the role of erythropoietin for neuronal and oligodendroglial differentiation in the hippocampus. Molecular Psychiatry, 21(12), 1752-1767.

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Immunofluorescence Imaging of Podocytes

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Human podocytes that were grown on type I collagen-coated glass cover slips were incubated at 37℃ for 2 h and fixed in 4% paraformaldehyde for 20 min. The cells were then permeabilized in 0.1% tritonX-100 for 10 min, blocked with 10% FBS for 30 min, washed three times for 5 min in phosphate buffered saline (PBS), and labeled with monoclonal rabbit anti-ZO-1 antibody (Invitrogen, Eugene, OR, USA). Phalloidin-FITC (Sigma-Aldrich Inc.) was utilized to stain F-actin. Primary antibody-bound specimens were incubated with 1:1000 (v/v) Alexa 594 for red conjugates and Alexa 488 for green (Invitrogen), respective of secondary anti-rabbit IgG, at room temperature for 40 min and at 37℃ for 20 min without CO₂. Nuclei were stained with 4'-6-diamidino-2-phenylindole (DAPI) (1:1000) for 20 min in PBS. Coverslips were mounted in aqueous mountant and viewed with a fluorescence microscope (Leica TCS SP2 AOBS, Mannheim, Germany).

Park S.J., Saleem M.A., Nam J.A., Ha T.S, & Shin J.I. (2015). Effects of Interleukin-13 and Montelukast on the Expression of Zonula Occludens-1 in Human Podocytes. Yonsei Medical Journal, 56(2), 426-432.

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FISH Analysis of NONMMUT100923.1 and miR-200a-3p

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FISH was conducted to detect the localization of NONMMUT100923.1 and miR-200a-3p in MEPS epithelial cells from the normal group (n = 3). Briefly, tissue slides (3-mm thick) of the embryonic head were lysed and incubated with Triton X-100 (Gibco, Grand Island, NY, USA) for 37 °C 20 min. The slides were hybridized with the denatured probe overnight at 27 °C. After overnight incubation, 3% bovine serum albumin blocking solution was added and incubated with the slides for 60 min, after which DAPI was added for 5 min. Anti-fluorescence stain was added to the slides, and staining was observed under a laser scanning co-aggregation microscope (TCS SP2 AOBS; Leica, Wetzlar, Germany).

Zhang M., Zhou J., Ji Y., Shu S., Zhang M, & Liang Y. (2023). LncRNA-NONMMUT100923.1 regulates mouse embryonic palatal shelf adhesion by sponging miR-200a-3p to modulate medial epithelial cell desmosome junction during palatogenesis. Heliyon, 9(5), e16329.

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Fixation and Staining of Embryos

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Relevant stages were fixed in 3.7% formalin-PBS for 15 to 30 min. After fixation, the eggs were rinsed in PBS and transferred to absolute methanol for storage at −8°C. Following washing steps in PBS the embryos were stained in either Hoechst (H33258, Molecular Probes) for 15 min or in Sytox®Green (Molecular Probes) for about 3 hours. The embryos were mounted in DABCO-glycerol and analyzed with a Zeiss Axioskop II and Leica TCS SP2 AOBS. The image stacks of the confocal laser scanning microscopy were further analyzed using the 3D image visualization software Imaris 5.0.3. (Bitplane AG, Zürich).

Alwes F, & Scholtz G. (2014). The early development of the onychopod cladoceran Bythotrephes longimanus (Crustacea, Branchiopoda). Frontiers in Zoology, 11, 10.

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HeLa Cell Matrigel Embedding and Imaging

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HeLa cells were embedded in Matrigel. To this end, 2 × 10⁵ cells/ml were suspended with Matrigel solution (21 mg/ml) to achieve a final concentration of ∼7 mg/ml at 4°C. 300 µl of this cell suspension was seeded in an 8-well chamber slide (Lab-Tek) and allowed to solidify at 37°C for 4 h. Cells were fixed in 4% paraformaldehyde and stained with anti-RAB5 Ab and phalloidin to detect F-actin. Sequential z sections (at least 30) of embedded cells were obtained by indirect immunofluorescence microscopy using a confocal microscope (TCS SP2 AOBS; Leica).

Frittoli E., Palamidessi A., Marighetti P., Confalonieri S., Bianchi F., Malinverno C., Mazzarol G., Viale G., Martin-Padura I., Garré M., Parazzoli D., Mattei V., Cortellino S., Bertalot G., Di Fiore P.P, & Scita G. (2014). A RAB5/RAB4 recycling circuitry induces a proteolytic invasive program and promotes tumor dissemination. The Journal of Cell Biology, 206(2), 307-328.

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Visualizing Phagosome Formation in BMDMs

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BMDMs were incubated with HZ (100 µM) in a 35-mm glass bottom tissue culture dish (MatTek) for 24 h. Cell membranes and DNA were stained with CellMask (red) and DAPI (blue), respectively. Confocal reflection microscopy (to visualize HZ phagocytosed by BMDM) combined with fluorescence microscopy was used as described in detail in [26 (link)], on a Leica TCS SP2 AOBS confocal laser-scanning microscope.

Corbett Y., D’Alessandro S., Parapini S., Scaccabarozzi D., Kalantari P., Zava S., Giavarini F., Caruso D., Colombo I., Egan T.J, & Basilico N. (2018). Interplay between Plasmodium falciparum haemozoin and l-arginine: implication for nitric oxide production. Malaria Journal, 17, 456.

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Transient Expression of MsTHI1-GFP Fusion

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The coding region of MsTHI1 without a stop codon was amplified using specific primers (MsTHI1-BamHI-F/MsTHI1-SacI-R, Supplementary Table S1) with BamHI and SacI restriction sites. The amplicons were double-digested with SacI and BamHI, followed by joining with pCAMBIA-1300 to create a fusion plasmid, pCAMBIA1300-MsTHI1-green fluorescent protein (GFP). The MsTHI1 fusion protein was transformed into the epidermal cells of N. benthamiana through the infiltration of Agrobacterium tumefaciens. A confocal laser scanning microscope was used to identify the fluorescence signal after 48 h incubation at 25°C in the dark (Leica TCS SP2 AOBS, Germany).

Yin H., Wang Z., Li H., Zhang Y., Yang M., Cui G, & Zhang P. (2022). MsTHI1 overexpression improves drought tolerance in transgenic alfalfa (Medicago sativa L.). Frontiers in Plant Science, 13, 992024.

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Monitoring Nuclear Calcium Spiking

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Assays for nuclear Ca²⁺ spiking were performed using young lateral root segments (0.5 to 0.8 cm long) freshly excised from hydroponically-grown transgenic plants expressing the p35S:NUP-YC2.1 cameleon reporter. Root explants were placed in a microchamber and treated with 150 μl of the solution to be tested as described in [18 (link)]. Confocal FRET-based ratio imaging for detecting and plotting relative changes of nuclear Ca²⁺ levels corresponding to YFP-to-CFP fluorescence intensity changes over time was performed according to [24 (link)] using a Leica TCS SP2 AOBS confocal laser-scanning microscope. Imaging was performed on both atrichoblasts and trichoblasts (root hairs) in the case of C. glauca. In contrast, Ca²⁺ spiking responses were only studied in D. trinervis atrichoblasts since few root hairs developed on D. trinervis roots under our growth conditions. It should be underlined that despite limited root hair formation on hydroponically-grown D. trinervis roots, efficient intercellular Frankia colonization and nodulation can be observed on these roots [e.g. 25]. Experiments were repeated several times for each treatment using independent roots (detailed in S1 Table).

Chabaud M., Fournier J., Brichet L., Abdou-Pavy I., Imanishi L., Brottier L., Pirolles E., Hocher V., Franche C., Bogusz D., Wall L.G., Svistoonoff S., Gherbi H, & Barker D.G. (2019). Chitotetraose activates the fungal-dependent endosymbiotic signaling pathway in actinorhizal plant species. PLoS ONE, 14(10), e0223149.

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