Lsm700 fluorescence microscope

Manufactured by Zeiss

Sourced in Germany

The LSM700 is a fluorescence microscope manufactured by Zeiss. It is designed for confocal imaging and analysis of fluorescently labeled samples. The microscope features high-resolution optical components and a flexible configuration to accommodate a variety of sample types and research applications.

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Lab products found in correlation

Mitotracker red, by Thermo Fisher Scientific (3 mentions) 4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (2 mentions) Triton, by Merck Group (2 mentions) Lb985 nightshade, by Berthold Technologies (1 mentions) Anti cd66b, by BioLegend (1 mentions) M087629 2, by Agilent Technologies (1 mentions) Fluoromount, by Zeiss (1 mentions) Alexa fluor 488 conjugated secondary antibody, by Thermo Fisher Scientific (1 mentions) Cell culture slide, by SPL Life Sciences (1 mentions) Deadend fluorometric tunel system, by Promega (1 mentions) Alexa fluor 555 labeled goat anti mouse igg, by Beyotime (1 mentions) Anti cdx2 antibody, by BioGenex (1 mentions) Triton x 100, by Beyotime (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Beyotime (1 mentions) Alexa fluor 568 donkey anti rabbit igg, by Thermo Fisher Scientific (1 mentions) H 1000, by Zeiss (1 mentions) Anti c ebpα, by Abcam (1 mentions) Prolong gold antifade reagent with dapi, by Thermo Fisher Scientific (1 mentions) Bovine serum albumin (bsa), by Merck Group (1 mentions) Anti e cadherin, by Abcam (1 mentions)

Close spelling variants of this product

LSM700 Laser Scanning Confocal Fluorescence microscope Fluorescence microscope

13 protocols using lsm700 fluorescence microscope

LCI and BiFC Assays in N. benthamiana

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The coding sequences of Dek701 and ZmRPABC2 were cloned into pCAMBIA-nLUC and pCAMBIA-cLUC constructs, respectively. The LCI assay was performed in N. benthamiana leaves by Agrobacterium-mediated infiltration (39 (link),40 (link)). Two days after infiltration, Luciferin (100 mM) was spread on the back of leaves for 15 min under dark condition, and luciferase activity was analyzed using a LB985 NightSHADE (Berthold Technologies, Bad Wildbad, Germany).
BiFC was performed in N. benthamiana leaves according to a previous work (41 (link)). The coding sequences of Dek701 and ZmRPABC2 were individually cloned into the pEarleygate201 and pEarleygate202 vectors harboring the nYFP and cYFP sequences, respectively, and co-infiltrated into N. benthamiana leaves through Agrobacterium-mediated transient infiltration. YFP fluorescence was detected after 48 h using a Zeiss LSM700 fluorescence microscope. Primers used for cloning of LCI and BiFC constructs are listed in Supplemental Data Set S1.

Chen Q., Guo Y., Zhang J., Zheng N., Wang J., Liu Y., Lu J., Zhen S., Du X., Li L., Fu J., Wang G., Gu R., Wang J, & Liu Y. (2023). RNA polymerase common subunit ZmRPABC5b is transcriptionally activated by Opaque2 and essential for endosperm development in maize. Nucleic Acids Research, 51(15), 7832-7850.

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Multicolor Immunofluorescence Tissue Staining

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Formalin-fixed and paraffin-embedded tissue sections were incubated with anti-CD66b (305102, Biolegend), M30 (12140322001, Roche) and anti-CD68 (M087629-2, Dako) primary antibodies, followed by secondary antibody staining using donkey anti-mouse IgG-AF555 (A31570, Thermo Fisher Scientific) and goat anti-mouse IgM-DyLight650 (SA5-10153, Thermo Fisher Scientific). Nuclei were stained with DAPI (33342, Invitrogen) and sections were visualised using a LSM700 fluorescence microscope (Zeiss, Vienna, Austria). For extended protocol, see supplementary material.

Schimek V., Strasser K., Beer A., Göber S., Walterskirchen N., Brostjan C., Müller C., Bachleitner-Hofmann T., Bergmann M., Dolznig H, & Oehler R. (2022). Tumour cell apoptosis modulates the colorectal cancer immune microenvironment via interleukin-8-dependent neutrophil recruitment. Cell Death & Disease, 13(2), 113.

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Immunofluorescence Staining of Cells

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Cells grown on coverslips were fixed with 4% paraformaldehyde in phosphate-buffered saline (PBS) for 10 min at room temperature, washed twice with PBS, and incubated for 16 h at 4°C with antibodies to FLAG (Merck) diluted in PBS containing 0.5% Triton X-100. They were then washed three times with PBS, incubated for 1 h at room temperature in the dark with Alexa Fluor 488–conjugated secondary antibodies (Thermo Fisher Scientific) diluted in PBS containing 0.5% Triton X-100, and then incubated for 5 min at room temperature in the dark with PBS containing DAPI. After two washes with PBS, the coverslips were mounted in Fluoromount and examined with an LSM 700 fluorescence microscope (Zeiss).

Nita A., Matsumoto A., Tang R., Shiraishi C., Ichihara K., Saito D., Suyama M., Yasuda T., Tsuji G., Furue M., Katayama B., Ozawa T., Murata T., Dainichi T., Kabashima K., Hatano A., Matsumoto M, & Nakayama K.I. (2021). A ubiquitin-like protein encoded by the “noncoding” RNA TINCR promotes keratinocyte proliferation and wound healing. PLoS Genetics, 17(8), e1009686.

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Transient Expression Assay in N. benthamiana

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With Nicotiana benthamiana, the transient expression assay was performed as according to Sparkes [68 (link)] with minor modification. Strains preparation: streaking the corresponding agrobacterium strains on LB plates, incubate at 28 °C for 2 days. A single strain was selected and transferred into 3 mL liquid LB medium with 8 h shaking. This was followed by subsequent shaking in 50 mL LB until OD₆₀₀ reached 0.6. Then, it was centrifuged at 3000 rpm and resuspended with infiltration buffer containing 10 mM 2-Morpholino ethanesulfonic acid hydrate (MES), 10 mM MgCl₂ and 100 µM acetosyringone to an OD₆₀₀ of 0.8, followed by incubating at 28 °C for 2 h. Infiltration: tobaccos leaves were injected from the back with a syringe (needle removed) until the water spots spread all over them. After 3 days, the YFP fluorescence intensity was detected with the LSM700 Fluorescence microscope (Zeiss, Oberkochen, Germany). Phytohormone treatment: tobacco leaves were treated with distilled water (CK), 50 µM MeJA or 25 µM abscisic acid (ABA) at 12 h after the infiltration, and the YFP signal was detected at 36 h after treatment.

Xiong J., Liu L., Ma X., Li F., Tang C., Li Z., Lü B., Zhou T., Lian X., Chang Y., Tang M., Xie S, & Lu X. (2020). Characterization of PtAOS1 Promoter and Three Novel Interacting Proteins Responding to Drought in Poncirus trifoliata. International Journal of Molecular Sciences, 21(13), 4705.

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Immunofluorescence Staining of Cells

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Cells on glass coverslips in 6-well plates were fixed in 4% polyformaldehyde for 10 min at room temperature, followed by membrane permeation using 100 μg/mL digitonin in PBS for 5 min. Cells were blocked in 3% BSA (bovine serum albumin) before primary antibodies were applied. Cells were incubated with primary antibodies in a moist container at room temperature for 1 h. Then, secondary antibodies conjugated with Alex-594 (ZSGB-BIO, Beijing,China) were applied to the cells in a moist container at room temperature for 1 h. The DNA in the nuclei was stained with 4′,6-diamidino-2-phenylindole (DAPI) at the final preparation step. The slides were viewed on an LSM700 fluorescence microscope (Zeiss, Oberkochen, Germany) using a 63× oil objective and laser at 488 or 594 nm for excitation.

Yang H., Jiang T., Fan L, & Qiu X. (2023). lncRNA LINC00960 promotes apoptosis by sponging ubiquitin ligase Nrdp1-targeting miR-183-5p: LINC00960 promotes apoptosis by Nrdp1/miR-183-5p. Acta Biochimica et Biophysica Sinica, 55(1), 91-102.

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Quantifying Apoptosis in N2a Cells

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To analyze apoptosis, N2a cells were seeded on a cell culture slide (SPL Life Sciences, Gyeonggi-do, Korea) and OGD and PDRN treatments were performed as previously described above. The DeadEnd™ Fluorometric TUNEL System (Promega Madison WI USA) was used to assess apoptosis according to the manufacturer’s protocol. Briefly, the samples were mounted on glass slides with a fluorescent mounting medium with DAPI for imaging using an LSM 700 fluorescence microscope (Carl Zeiss, Gottingen, Germany). The number of positively stained cells over the total number of cells per specimen field was measured, and the percentage of positive cells was calculated. Four individual specimens were analyzed per group.

Jo S., Baek A., Cho Y., Kim S.H., Baek D., Hwang J., Cho S.R, & Kim H.J. (2023). Therapeutic effects of polydeoxyribonucleotide in an in vitro neuronal model of ischemia/reperfusion injury. Scientific Reports, 13, 6004.

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Embryonic CDX2 Expression Imaging

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Embryos were fixed overnight at 4°C in immunostaining fixing solution (Beyotime, P0098). Unless otherwise stated, all staining steps were performed at room tem-perature. Embryos were permeabilized with immunostaining solution containing Triton X-100 (Beyotime, P0096) for 0.5 h. After washing 5 times, they were blocked at 4°C for 12 h in the immunostaining blocking solution (Beyotime, P0260) and then incubated with primary antibody at 4°C for 12 h. Anti-CDX2 antibody (BioGenex) was diluted 1:50 using QuickBlock dilution buffer for immunostaining (Beyotime, P0262). After washing 5 times, embryos were incubated with secondary antibody Alexa Fluor 555-labeled goat antimouse IgG (Beyotime, A0459) for 2 h, washed, and then sealed with 4,6-diamidino-2-phenylindole (DAPI; Beyotime, C1005), 10 min, sealing tablets (Su et al., 2012) (link). Finally, the embryos were photographed with a LSM 700 fluorescence microscope (Carl Zeiss). All experiments were replicated at least 3 times with a group of 10 to 15 embryos in each replicate. For fluorescence quantification, the intensity levels of the NT group and ZFP57 overexpression/knockdown embryos were expressed relative to the average intensity level of IVF embryos. Intensities were measured as described previously (Zhang et al., 2022b) .

Yu T., Meng R., Song W., Sun H., An Q., Zhang C., Zhang Y, & Su J. (2023). ZFP57 regulates DNA methylation of imprinted genes to facilitate embryonic development of somatic cell nuclear transfer embryos in Holstein cows. Journal of dairy science, 106(1).

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Activated Relish Immunofluorescence Staining

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For immunofluorescent staining of the activated Relish, polyserum against Relish (He et al., 2017) was raised in rabbits (Abmart, CAN). Fat body samples were washed in PBS and fixed in 4% paraformaldehyde for 20 min. The samples were washed three times with PBS (5 min each wash) and then incubated with anti-Relish antiserum (7 mg/mL) in PBST containing 0.5% BSA at 4 C for 16 h. After being washed five times with PBS, the tissue was further incubated with 2 mg/mL Alexa Fluor 568 donkey anti-rabbit IgG (Invitrogen) for 1 h in the dark. The samples were mounted (VETASHIELD, H-1000), and imaged by a LSM700 fluorescence microscope (Carl Zeiss) and analyzed by the ZEN software.

Yang S., Zhao Y., Yu J., Fan Z., Gong S.T., Tang H, & Pan L. (2019). Sugar Alcohols of Polyol Pathway Serve as Alarmins to Mediate Local-Systemic Innate Immune Communication in Drosophila. Cell host & microbe, 26(2).

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Immunofluorescence Assay for Cell Characterization

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Cells were cultured on microscope glasses (Sigma-Aldrich). Coverslips were washed twice with PBS and fixed using PBS containing 4% paraformaldehyde (Merck) for 20 min at room temperature. Cells were washed twice with PBS and permeabilized with PBS containing 0.25% Triton (Sigma) for 5 min at room temperature. Posteriorly, cells were pre-incubated with PBS containing 2% of bovine serum albumin (Sigma) for 60 min at room temperature. After the blocking step, cells were incubated overnight at 4° with anti-C/EBPα (Abcam, ab128482), anti-E-cadherin (Abcam, ab1416), and anti-Fibronectin (BD Biosciences, 610077). Cells were washed twice in PBS, and respective secondary antibodies (BD Biosciences) were incubated for 60 min, at room temperature. Lastly, cells were washed twice in PBS and coverslips were mounted in Prolong Gold anti-fade reagent with DAPI (Invitrogen). Confocal images were acquired using a Zeiss LSM 700 fluorescence microscope (Zeiss).

Lourenço A.R., Roukens M.G., Seinstra D., Frederiks C.L., Pals C.E., Vervoort S.J., Margarido A.S., van Rheenen J, & Coffer P.J. (2020). C/EBPɑ is crucial determinant of epithelial maintenance by preventing epithelial-to-mesenchymal transition. Nature Communications, 11, 785.

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Fluorescent Imaging of Mitochondrial Proteins

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HepG2, 293T, and HaLa cells were seeded onto coverslips which coated with poly‐L‐lysine and fixed with 4% paraformaldehyde after transfection for 48 h. The cells were treated with Triton‐X‐100 (Sigma Aldrich, St. Louis, MO, Cat # V900502), washed with PBS and blocked with bovine serum albumin followed by incubated with anti‐mouse‐DNMT1 antibody (Abcam, Cambridge, MA) and anti‐rabbit‐VDAC antibody (Cell Signaling Technology, Danvers, MA). Then the cells were incubated with Alexa Fluor 555‐conjugated anti‐mouse IgG and Alexa Fluor 488‐conjugated anti‐rabbit IgG (Invitrogen, Carlsbad, CA) antibodies. After washing with PBS, the cells were stained with DAPI (Invitrogen, Carlsbad, CA). Images were visualized with a ZEISS LSM700 fluorescence microscope at 63x magnification (Carl Zeiss, Chicago, IL). For detection of ND6 siRNA mitochondrial localization, mouse ND6 siRNA with FAM modification was used to transfect 3T3‐L1 cells for 48 h. Then the cells were stained with Mito Tracker Red (Thermo‐Fisher, Waltham, MA, Cat # M7512) to mark mitochondrial morphology. For detection of mitochondrial superoxide, 3T3‐L1 and HT22 cells were transfected with mouse ND6 siRNA for 48 h, and then stained with MitoSox Red (Thermo‐Fisher, Waltham, MA, Cat # M36008).

Cao K., Lv W., Wang X., Dong S., Liu X., Yang T., Xu J., Zeng M., Zou X., Zhao D., Ma Q., Lin M., Long J., Zang W., Gao F., Feng Z, & Liu J. (2021). Hypermethylation of Hepatic Mitochondrial ND6 Provokes Systemic Insulin Resistance. Advanced Science, 8(11), 2004507.

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