Confocal microscope lsm700

Manufactured by Zeiss

Sourced in Germany

The Zeiss LSM700 is a confocal microscope designed for high-resolution imaging. It features a laser-scanning system and advanced optics for capturing detailed, three-dimensional images of samples. The LSM700 is capable of performing fluorescence imaging and can be used for a variety of applications in life science research and materials science.

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Lab products found in correlation

Zen black software, by Zeiss (2 mentions) Live dead baclight kit, by Thermo Fisher Scientific (1 mentions) Zen 2012, by Zeiss (1 mentions) Ultracruz aqueous mounting medium with dapi, by Santa Cruz Biotechnology (1 mentions) 8 well chamber slide, by Sarstedt (1 mentions) Alexa fluor 488, by Thermo Fisher Scientific (1 mentions) Bovine serum albumin (bsa), by Merck Group (1 mentions) Triton x 100, by Cell Biolabs (1 mentions) Pe conjugated anti cd4, by BioLegend (1 mentions) Apc conjugated anti b220, by Thermo Fisher Scientific (1 mentions) Fluoromount g mounting solution, by Southern Biotech (1 mentions) 80i fluorescence microscope, by Nikon (1 mentions) Alexa488 coupled antibodies, by Thermo Fisher Scientific (1 mentions) Anti cith3, by Abcam (1 mentions) Vectashield antifade mounting medium for fluorescence, by Vector Laboratories (1 mentions)

Close spelling variants of this product

LSM700 Meta confocal laser scanning microscope LSM700 confocal laser microscope LSM700 Confocal Laser Scanning Microscope system LSM700 upright confocal microscope LSM700 Laser Scanning Confocal Fluorescence microscope Carl Zeiss LSM700 confocal microscope LSM700 Axio Examiner Z.1 confocal microscope Observer Z1 LSM700 confocal microscope LSM700 inverted laser confocal microscope Confocal Laser Scanning Microscope LSM700

11 protocols using confocal microscope lsm700

Subcellular Localization of siRNA-Loaded GARP iLNPs

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To study the subcellular localization of siRNA‐loaded GARP iLNPs, the HepG2‐luc cells were seeded into 35 mm dishes at a density of 2 × 10⁵ cells per well. After 24 h, cells were transfected with Cy5‐siRNA‐loaded GAP35 iLNPs and GARP35 iLNPs at the final siRNA concentrations of 50 and 150 nM, respectively. Four hours later, the cells were washed three times with 1× PBS and stained with 1 µL of Hoechst 33342 solution (1 mg/mL in PBS, for staining nucleus) and 0.3 µL of LysoTracker Green (1: 3000 in PBS, for staining endosome and lysosome). After staining for 30 min, cells were washed twice with 1× PBS and then recorded with Zeiss confocal microscope (LSM700, Carl Zeiss, Germany).
In order to analyze the co‐localization of siRNA‐loaded GARP iLNPs with the endosomes or lysosomes at different transfection time points, HepG2‐luc cells were plated into 35 mm dishes at a density of 2 × 10⁵ cells per well. After 24 h, the siRNA‐loaded GARP iLNPs at the final siRNA concentration of 50 nM was added to the dishes. Lipo/Cy5‐siRNA NPs served as a control group. Then the cells were stained and imaged according to above‐mentioned protocol at 1, 3, 5, 8, 10, and 12 h after transfection.

Guo S., Li K., Hu B., Li C., Zhang M., Hussain A., Wang X., Cheng Q., Yang F., Ge K., Zhang J., Chang J., Liang X., Weng Y, & Huang Y. (2021). Membrane‐destabilizing ionizable lipid empowered imaging‐guided siRNA delivery and cancer treatment. Exploration, 1(1), 35-49.

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Visualizing Intracellular Salmonella in Caco-2 Cells

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Confocal microscopic examination was performed to visualize intracellular S. Typhimurium in Caco-2 cells pretreated with 25 and 50 μM of Rg3 [27 ]. Briefly, Caco-2 cells (10⁴/mL) were seeded onto chambered glass slides in antibiotic-free MEM. After incubation at 37°C for 24 h, cells were treated and infected by a method similar to the invasion assay described previously. Then, the medium was aspirated and cells were washed twice with 3-(N-morpholino) propanesulfonic acid (0.1 M MOPS, pH 7.2) buffer mixed with magnesium chloride (MgC_l2, 1 mM) (MOPS/MgC_l2). The medium was replaced by 400 μL of BacLight staining solution (Invitrogen, Thermo Fisher Scientific, Eugene, OR, USA) containing 5 μM SYTO9 and 30 μM propidium iodide mixed with saponin (0.1%) in MOPS/MgC_l2. After 15–30 min of incubation at room temperature, cells were washed (2 × ) in MOPS/MgC_l2 and covered with coverslips. Finally, slides were examined using a Zeiss confocal microscope (LSM700; Carl-Zeiss, Jena, Germany).

Mechesso A.F., Quah Y, & Park S.C. (2019). Ginsenoside Rg3 reduces the adhesion, invasion, and intracellular survival of Salmonella enterica serovar Typhimurium. Journal of Ginseng Research, 45(1), 75-85.

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Subcellular Localization of NPs/siRNA

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For studies of the subcellular localization of NPs/siRNA complexes, MDA-MB-231-Luc cells were seeded into 35mm dishes at 2×10⁵ cells each well. Twenty-four hours later, cells were transfected with PTMS/Cy5-siRNA at the final siRNA concentration of 50nM. After 4h and 10h, cells were washed three times with 1×PBS and stained by Lysotracker Green (indicated endosome/lysosome). Then cells were fixed by paraformaldehyde (4%) for 40min and stained by DAPI (indicated nucleus). At last, cells were imaged by Zeiss confocal microscope (LSM700, Carl Zeiss, Germany).

Du L., Zhou J., Meng L., Wang X., Wang C., Huang Y., Zheng S., Deng L., Cao H., Liang Z., Dong A, & Cheng Q. (2017). The pH-Triggered Triblock Nanocarrier Enabled Highly Efficient siRNA Delivery for Cancer Therapy. Theranostics, 7(14), 3432-3445.

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Membrane Integrity Assay for S. Typhimurium

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Membrane integrity was determined by using the LIVE/DEAD BacLight kit (Molecular Probes, Eugene, OR, USA) with a slight modification of the manufacturer’s instructions. S. Typhimurium (10⁶ CFU/mL) was treated as described in the time–kill assay, for 4 h. Three microliters of SYTO9 and a propidium iodide (PI) mixture (1:1) were added to 1 mL of pre-treated bacterial cultures and incubated in the dark for 15 min. Samples were then visualized using a Zeiss confocal microscope (LSM700, Carl-Zeiss, Jena, Germany) at excitation/emission wavelengths of 480/500 and 490/635 nm for SYTO9 and PI stain, respectively.

Mechesso A.F., Yixian Q, & Park S.C. (2019). Methyl gallate and tylosin synergistically reduce the membrane integrity and intracellular survival of Salmonella Typhimurium. PLoS ONE, 14(9), e0221386.

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Immunofluorescence of FFAR4 in Oocytes

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Immature oocytes with a partially reduced cumulus were fixed for 30 min in 4% paraformaldehyde diluted in PBS (phosphate buffer saline) pH 7.4. Following 15 min of incubation in 0.1% Triton/PBS, oocytes were incubated in PBS containing 5% goat serum and 2% bovine serum albumin (BSA). Samples were then incubated at 4 °C overnight either with rabbit FFAR4 antibody (9.5 ng/mL) or pre-immune serum (similar IgG concentration) of the same rabbit (negative control), both diluted in PBS/0.5% BSA. After washing four times for 30 min in PBS/0.1% BSA, the oocytes were incubated for 3 h in the dark with the secondary antibody, Alexa Fluor488 conjugated goat anti-rabbit IgG. Another four 30 min washes with PBS/0.1% BSA were performed, then OCC were incubated with Hoechst 33,258 (1 μg/mL) for 15 min and mounted on slides using Moviol® mounting medium. Green (Alexa 488) and blue (Hoechst) fluorescence was observed with a Zeiss confocal microscope (LSM700; Carl Zeiss Microscopy GmbH, Munich, Germany) using a 20× objective or an oil 63× objective and the appropriate filters. The images were collected with Zen 2012 software (black edition version 8.0, Carl Zeiss Microscopy GmbH).

Elis S., Oseikria M., Vitorino Carvalho A., Bertevello P.S., Corbin E., Teixeira-Gomes A.P., Lecardonnel J., Archilla C., Duranthon V., Labas V, & Uzbekova S. (2017). Docosahexaenoic acid mechanisms of action on the bovine oocyte-cumulus complex. Journal of Ovarian Research, 10, 74.

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Subcellular Localization of Dually Labeled Nanoassemblies

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For subcellular localization experiments, a dually labeled assembly
was used with ASO labeled with Cy5.5 at the 5′ end and rHA
labeled with Cy3. C28/I2 cells were seeded overnight at 10,000 cells/well
in an 8-well chamber slide (Sarstedt, cat# 94.6140.802) at 37 °C,
5% CO₂. The cells were then treated with the dually labeled
assembly at 100 nM and incubated for further 4, 6, and 24 h at 37
°C, 5% CO₂. Cells were washed with DPBS and then fixed
with formalin 10% (Sigma-Aldrich, cat# HT5014) for 20 min. After further
washing, cells were permeabilized with 0.1% Triton X-100 (Cell Biolabs,
INC., cat# 124102) for 20 min, followed by blocking with 5% bovine
serum albumin (BSA, Sigma-Aldrich, cat# A9647) in DPBS for 1 h. Cells
were washed and incubated with primary rabbit polyclonal antibodies
(Abcam), (anti-EEA1, cat# ab2900, for early endosomes, and anti-Lamp1,
cat# ab24170, for lysosomes) in 5% BSA at 4 °C overnight. The
cells were then washed and incubated with goat antirabbit secondary
antibody, Alexa Fluor 488 (Invitrogen, cat# A11034) for 90 min, followed
by washing and mounting the slide with UltraCruz Aqueous Mounting
Medium with DAPI (Santa Cruz Biotechnology, Inc., cat# sc-24941).
The slides were kept at 4 °C in the dark until visualization
on a Zeiss Confocal microscope LSM 700 (Carl Zeiss MicroImaging).
Images were processed with Zeiss Zen Black 2012 edition.

Elkhashab M., Dilek Y., Foss M., Creemers L.B, & Howard K.A. (2024). A Modular Albumin-Oligonucleotide Biomolecular Assembly for Delivery of Antisense Therapeutics. Molecular Pharmaceutics, 21(2), 491-500.

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Multimodal Immune Profiling of Tissue

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Th17 (CD4, IL-17), Treg (CD4, CD25, Foxp3), SMILE, AMPK, mTOR, pSTAT3 705, pSTAT3 727 and germinal center markers (CD3, B220, PNA) expression levels were analyzed by confocal microscopy. Tissue Sects. (7 μm thick) were fixed in methanol-acetone and stained with phycoerythrin (PE)-conjugated anti-CD4 (Biolegend, San Diego, CA, USA), fluorescein isothiocyanate (FITC)-conjugated anti CD4 (BD Bioscience, San Diego, CA, USA), PE-conjugated anti-interleukin-17 (eBioscience, San Diego, CA, USA), allophycocyanin (APC)-conjugated anti-CD25 (Biolegend), PE-conjugated anti-Foxp3 (Thermo Fisher Scientific, Waltham, MA, USA), PE-conjugated anti-phosphorylated Stat3 (Tyr 705, Ser 727, BD Bioscience), APC-conjugated anti-B220 (eBioscience), Alexa 594-conjugated anti-PNA (Thermo Fisher Scientific), SMILE (Abcam, Cambridge, UK), AMPK (Abcam) and mTOR (Cell Signaling). After overnight incubation at 4℃, the stained sections were analyzed on a Zeiss confocal microscope (LSM 700; Carl Zeiss, Oberkochen, Germany) at 200× magnification and Zen Blue edition software.

Jhun J., Moon J., Kwon J.Y., Cho K.H., Lee S.Y., Na H.S., Cho M.L, & Min J.K. (2023). Small heterodimer partner interacting leucine zipper protein (SMILE) ameliorates autoimmune arthritis via AMPK signaling pathway and the regulation of B cell activation. Cell Communication and Signaling : CCS, 21, 98.

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Brain Tissue Preparation and Immunohistochemistry

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At 6 months after transplantation, animals were perfused transcardially with 4% paraformaldehyde. The brain was serially sectioned coronally and sagittally to 35 μm in thickness and subsequently stored in the preservation solution at −20°C.
Immunohistochemistry was performed as previously described (Liu et al., 2013b (link)). Sections were washed with PBS for 5 min three times, and permeabilized and blocked for 1 hr in 10% donkey serum and 0.2% Triton X-100 before being incubated in the primary antibody in 5% serum and 0.2% Triton X-100 at 4°C overnight. Sections were subsequently washed and stained with Alexa Fluor (Life) secondary antibodies and Hoechst in 5% donkey serum for 1 hr before being washed and mounted onto glass slides with Fluoromount-G mounting solution (SouthernBiotech). The primary antibodies used in this study are listed in Table S2. Images were visualized using a Nikon 80i fluorescence microscope (Nikon Instruments) and a Zeiss confocal microscope (LSM700, Zeiss instruments).

Huo H.Q., Qu Z.Y., Yuan F., Ma L., Yao L., Xu M., Hu Y., Ji J., Bhattacharyya A., Zhang S.C, & Liu Y. (2018). Modeling Down Syndrome with Patient iPSCs Reveals Cellular and Migration Deficits of GABAergic Neurons. Stem Cell Reports, 10(4), 1251-1266.

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Immunofluorescence Localization of Tribbles Proteins

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Bovine and murine COCs or CC after in vitro culture were fixed during 30 min in 4% paraformaldehyde solution in PBS (pH 7.4). After fixation, cell permeabilization was performed in PBS/BSA 0.5%/Triton X100 0.5% for 15 min. Cells were then incubated in PBS/BSA 2% supplemented with 5% goat or horse serum for 1 h to block non-specific binding sites. A wash with PBS/BSA 0.1%/Tween 20 0.1% preceded the overnight incubation with the primary antibodies at 4°C. Goat antibodies against recombinant human (rh) TRIB1 (Santa Cruz Biotechnologies, Heidelberg, Germany), rabbit rh TRIB2 antibody and rabbit rh TRIB3 antibody (Sigma) were used at 1:100 dilution. Four 15-min washes preceded the incubation of the secondary donkey anti-goat or goat anti-rabbit Alexa488-coupled antibodies (Molecular Probes, Invitrogen) at room temperature for 2 h. Four PBS/BSA 0.1%/Tween 0.1% washes of 20 min were performed and 1 μg/mL of Hoechst was added in the last one. Lastly a drop of Mowiol® anti-fade solution was added to mount the slides and allow confocal microscopy analysis. Immunofluorescence was observed using a Zeiss confocal microscope LSM700 (Carl Zeiss Microscopy GmbH, Munich, Germany) using oil 40× and 63× objectives.

Brisard D., Chesnel F., Elis S., Desmarchais A., Sánchez-Lazo L., Chasles M., Maillard V, & Uzbekova S. (2014). Tribbles expression in cumulus cells is related to oocyte maturation and fatty acid metabolism. Journal of Ovarian Research, 7, 44.

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Podocyte Cytoskeleton Dynamics Imaging

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For LifeAct experiments, podocytes overexpressing LifeAct-GFP, MC1R-mCherry or E92K-MC1R-mCherry were used. The cells were placed on a Zeiss Confocal LSM700 Microscope (Carl Zeiss AB) and a total of 120 images were acquired with 30 sec intervals during 60 min using an x 40 lens and Zeiss ZEN Black software (Carl Zeiss AB). The images from the time-lapse series were exported and analyzed using computer-assisted image analysis with software from Visiopharm (Visiopharm). Visiopharm was used to analyze the photographs for calculation of the total podocyte area over time and compared to the 0-min time point.

Bergwall L., Wallentin H., Elvin J., Liu P., Boi R., Sihlbom C., Hayes K., Wright D., Haraldsson B., Nyström J, & Buvall L. (2018). Amplification of the Melanocortin-1 Receptor in Nephrotic Syndrome Identifies a Target for Podocyte Cytoskeleton Stabilization. Scientific Reports, 8, 15731.

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