Ti e a1

Manufactured by Nikon

Sourced in Japan

The Ti-E A1 is a high-performance inverted microscope system designed for advanced microscopy applications. It features a sturdy frame, stable focusing mechanism, and a variety of optical components to support various imaging techniques. The core function of the Ti-E A1 is to provide a reliable and versatile platform for researchers to conduct their microscopy studies.

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Lab products found in correlation

Na oil objective, by Nikon (2 mentions) Agonist acetylcholine, by Solarbio (2 mentions) Af dx 384, by Merck Group (2 mentions) Safire2, by Tecan (2 mentions) Ab150080, by Abcam (1 mentions) Alexa fluor 594 conjugated goat anti rabbit igg, by Cell Signaling Technology (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Solarbio (1 mentions) Anti rabbit igg fab2 alexa fluor r 488, by Cell Signaling Technology (1 mentions) Phalloidin ifluor 594, by Abcam (1 mentions) Dapi staining solution, by Beyotime (1 mentions) C6 flow cytometry, by BD (1 mentions) Flow cytometry, by Agilent Technologies (1 mentions) Vectashield mounting medium with dapi, by Vector Laboratories (1 mentions) In situ cell death detection kit, by Roche (1 mentions) Mc170 hd, by Leica camera (1 mentions) Ab92516, by Abcam (1 mentions) Ab175695, by Abcam (1 mentions) Ab217344, by Abcam (1 mentions) Ab79823, by Abcam (1 mentions) Celesta, by BD (1 mentions)

24 protocols using ti e a1

Investigating NF-κB p65 Translocation

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The NF-κB p65 translocation was investigated using immunofluorescence microscopy. Cells were seeded and pre-treated with different doses of xanthatin with or without LPS for 24 h in 12-well plates. The operation primarily refers to our group’s testing procedure [77 (link)]. Finally, the crawls were visualized by a laser scanning confocal microscope (Nikon Ti-EA1, Japan).

Liu Y., Chen W., Zheng F., Yu H, & Wei K. (2022). Xanthatin Alleviates LPS-Induced Inflammatory Response in RAW264.7 Macrophages by Inhibiting NF-κB, MAPK and STATs Activation. Molecules, 27(14), 4603.

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NF-κB Nuclear Translocation Inhibition by 5-HMF

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NF-κB nuclear translocation was examined by immunofluorescence microscopy. RAW 264.7 cells (5 × 10⁴ cells/each well in a 24-well plate with sterile coverslips) were pre-treated with 0, 31.5, 63.0 and 126.0 μg/mL of 5-HMF for 6 h and then stimulated by 1 μg/mL of LPS for 18 h. Cells were fixed with 4% paraformaldehyde for 15 min at room temperature and then washed with PBS twice. Later, the cells were washed with a quenching solution (0.1% glycine in PBS) twice, permeated with 0.1% Triton X-100 for 10 min and blocked using a blocking solution (10% FBS in PBS) for 1 h at room temperature. Then the cells were incubated with the primary p65 antibody at 4 °C overnight and later incubated with Conjugated Goat anti-Rabbit IgG H&L (Alexa Fluor^® 594) (1:2000, ab150080, Abcam, UK) for 1 h at room temperature and then stained with 4,6-diamidino-2 phenylindole (DAPI) for 5 min. The coverslips were removed and observed using laser scanning confocal microscope (Nikon Ti-E A1, Japan).

Kong F., Lee B.H, & Wei K. (2019). 5-Hydroxymethylfurfural Mitigates Lipopolysaccharide-Stimulated Inflammation via Suppression of MAPK, NF-κB and mTOR Activation in RAW 264.7 Cells. Molecules, 24(2), 275.

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Nanoscale Imaging Using Confocal Microscopy

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Nanochambers were prepared on No. 1.5H cover slips and images were acquired using an oil immersion objective (60×, N.A 1.4) on a Nikon Ti-E A1+ at a laser wavelength of 490 nm and emission bandpass of 510–560 nm with the pinhole set to 0.4 AU. The detector was a photomultiplier tube.

Svirelis J., Adali Z., Emilsson G., Medin J., Andersson J., Vattikunta R., Hulander M., Järlebark J., Kolman K., Olsson O., Sakiyama Y., Lim R.Y, & Dahlin A. (2023). Stable trapping of multiple proteins at physiological conditions using nanoscale chambers with macromolecular gates. Nature Communications, 14, 5131.

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Immunofluorescence Staining of Beta-Catenin

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Aggregates were collected, washed with PBS, and fixed overnight in 4% paraformaldehyde at 4 °C, and then permeabilized with 0.5% Triton X‐100 for 20 min at 4 °C. Three washes with PBS were included between each step. Following washing, the samples were incubated in blocking buffer containing goat serum for 30–60 min at room temperature, then incubated in PBS containing primary antibodies such as rabbit anti-beta catenin antibody (Bioss, bs-23663R, 1:500) overnight at 4 ℃ followed by rewarming to room temperature and incubation in PBS containing secondary antibodies such as Alexa Fluor 594-conjugated goat anti-rabbit IgG (Cell Signaling Technology, 8889S, 1:800) for 1 h in the dark at room temperature. Nuclei were counterstained by incubation with DAPI (Solarbio, C0065) for 5 min, and the fluorescence signal was imaged on the single photon confocal microscopy (Ti-E A1, Nikon). The details of all antibodies used in the present study are listed in Additional file 2: Table S2.

Wu H., Tang X., Wang Y., Wang N., Chen Q., Xie J., Liu S., Zhong Z., Qiu Y., Situ P., Zern M.A., Wang J., Chen H, & Duan Y. (2022). Dextran sulfate prevents excess aggregation of human pluripotent stem cells in 3D culture by inhibiting ICAM1 expression coupled with down-regulating E-cadherin through activating the Wnt signaling pathway. Stem Cell Research & Therapy, 13, 218.

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Immunofluorescence Imaging of Cells

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The non-polarized and polarized hEHs were fixed using 4% paraformaldehyde fix solution. After permeabilizing by 0.5% Triton and blocking by goat serum, the cells were incubated with primary antibody at 4 °C overnight. Then, the cells were co-incubated with anti-rabbit IgG Fab2 Alexa Fluor (R) 488 (Cell Signaling Technology, Boston, MA, USA) and Phalloidin-iFluor 594 (Abcam, Cambridge, UK). After washing, the cells were incubated with DAPI staining solution (Beyotime). After being thoroughly washed, the samples were removed from the hanging insert and placed on microscope slides. The fluorescence signal was imaged on the single photon confocal microscopy (Ti-E A1, Nikon, Tokyo Met. Japan). Antibodies used were listed in Table S1.

Wang J., Situ P., Chen S., Wu H., Zhang X., Liu S., Wang Y., Xie J., Chen H, & Duan Y. (2022). Hepatic Polarized Differentiation Promoted the Maturity and Liver Function of Human Embryonic Stem Cell-Derived Hepatocytes via Activating Hippo and AMPK Signaling Pathways. Cells, 11(24), 4117.

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Quantification of Muscarinic Receptor Activation

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In some experiments, the fluorescence signals of HEK293T cells transfected with the muscarinic receptor-based chimeric
constructs were measured with a TECAN Safire2 fluorescence plate reader (TECAN, Männedorf, Switzerland; excitation, 480 nm;
emission, 520 nm). During the measurement, the culture media was replaced with 100 μl Tyrode solution containing ACh at
varied concentrations from 0–100 μM. The ΔF/F₀ of each construct was obtained by averaging the
ACh-induced fluorescence responses of transfected wells after digitally subtracting that of neighboring control non-transfected
wells.
In other culture cell experiments, HEK293T cells and cultured neurons were imaged by an inverted Nikon Ti-E A1 confocal
microscope with a 40×/1.35 NA oil objective (Nikon, Tokyo, Japan). Cells were perfused with standard extracellular Tyrode
solution containing (in mM): 150 NaCl, 4 KCl, 2 MgCl₂, 2 CaCl₂, 10 HEPES and 10 Glucose, with pH of 7.4, in
an imaging chamber during imaging. Agonist acetylcholine (Solarbio, Beijing, China), tiotropium bromide (Dexinjia Bio & Tech
Co., Ltd, Jinan, China), and AF-DX 384 (Sigma-Aldrich) were delivered with a custom-made perfusion system and/or bath applied. The
chamber was washed with Tyrode solution between applications and cleaned with 75% ethanol between experiments.

Jing M., Zhang P., Wang G., Feng J., Mesik L., Zeng J., Jiang H., Wang S., Looby J.C., Guagliardo N.A., Langma L.W., Lu J., Zuo Y., Talmage D.A., Role L.W., Barrett P.Q., Zhang L.I., Luo M., Song Y., Zhu J.J, & Li Y. (2018). ED SUM: Signaling by the neurotransmitter acetylcholine is monitored in cells and animals with a sensitive reporter.: A genetically encoded fluorescent acetylcholine indicator for in vitro and in vivo studies. Nature biotechnology, 36(8), 726-737.

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Nanoparticle Cellular Uptake Kinetics

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Flow cytometry analysis: HaCaT cells were cultured at 1 × 10⁵ cells well⁻¹ in 12 well plates. When the cells properly adhered, the medium was removed and FITC-loaded PLGA nanoparticle suspension (100 µg ml⁻¹) was incubated for 1, 2, 4 and 8 h. After different exposure times, cells were washed twice with phosphate buffered saline (PBS) and harvested using trypsin–EDTA (0.25% v/v). The results were recorded using a BD C6 flow cytometry (FACS) with a 488 nm laser. For each experiment, a total of 10 000 cells were gated per sample and each sample was performed in triplicate.
Confocal laser scanning microscopy: laser confocal microscopy (Ti-E A1, Nikon, Japan) was used to qualitatively analyse the localization of fluorescent nanoparticles in HaCaT cells. The cells were seeded at 5 × 10⁴ cells well⁻¹ in 24-well plates containing glass coverslips. After incubation with fluorescent nanoparticles for a certain period of time, the cells were washed gently with PBS three times and fixed with 4% paraformaldehyde for 20 min. Finally, cells were stained with 4,6-diamidino-2 phenylindole (DAPI) for 5 min.

Hu F., Liu W., Yan L., Kong F, & Wei K. (2019). Optimization and characterization of poly(lactic-co-glycolic acid) nanoparticles loaded with astaxanthin and evaluation of anti-photodamage effect in vitro. Royal Society Open Science, 6(10), 191184.

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Confocal Microscopy Analysis of DOX Uptake

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Laser scanning confocal microscopy (CLSM, TI-E-A1, Nikon Corporation, Tokyo, Japan) was used to observe the fluorescence of Huh-7 cells. First, Huh-7 cells were added to 35 mm four-well culture dishes and incubated for 12 h at 37°C in a CO₂ incubator; the original culture medium was removed, and 500 μL of DP@ Fe₃O₄@DOX-FA or free DOX culture medium was added and placed in a 37°C CO₂ incubator for 4 h or 2 h with or without the AMF (418 Oe, 250 kHz, 10 min); the culture medium was removed and washed with PBS for three times (2 mL/time); 1 mL of 4% paraformaldehyde solution was added and placed in a refrigerator at 4°C for 12h, and then imaging was performed. Further, the cellular uptake of DOX was measured by flow cytometry at 488 nm (Agilent, Hangzhou, China). Briefly, Huh-7 cells were inoculated in 24-well plates at a 1.0×10⁵ cells/well. After 12 hours of cell culture, the cells were treated as described above. Subsequently removing the medium, cells were collected using tryptic digestion. Supernatants were then removed, and the cells were centrifuged again with 300ul stain buffer. The resulting mixture was filtered and added to flow-through tubes to detect DOX fluorescence. The data were analyzed using NovoExpress software.

Zhu J., Yang Y., Wang J., Hong W., Li Y., Wang Z, & Li K. (2023). Dual Responsive Magnetic Drug Delivery Nanomicelles with Tumor Targeting for Enhanced Cancer Chemo/Magnetothermal Synergistic Therapy. International Journal of Nanomedicine, 18, 7647-7660.

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Confirming Neuronal Morphology and Transduction

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ChR2-eYFP expression at the viral vector injection sites (IL and PL) and on the mPFC axons in the amygdala were verified as described before.^{16 (link)} To confirm the location and to visualize the morphology of recorded neurons, the recorded slices were fixed in 4% paraformaldehyde in 0.1M phosphate buffer (PB) for 12–24 h at 4°C. After fixation, slices were washed in phosphate buffed saline (PBS) (3 × 10 min), and permeabilized in PBS containing 0.2% Triton X-100 for 60 min. Slices were then incubated in fluorescently-conjugated streptavidin (1:300, Streptavidin, Alexa Fluor 594 conjugate, Life Technologies) for 12–24 h at 4°C. Finally, the slices were washed in PBS (3 × 10 min), mounted on slides with Vectashield mounting medium with DAPI (Vector Laboratories), and imaged under a confocal microscope (Ti-E, A1, Nikon).

Kiritoshi T, & Neugebauer V. (2018). Pathway-specific alterations of cortico-amygdala transmission in an arthritis pain model. ACS chemical neuroscience, 9(9), 2252-2261.

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Immunofluorescence Detection of His-Tagged Protein

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The expression of target protein labeled with 6 × His Tag in UCMSCs-Tandab(IL-6/CD20) was detected by immunofluorescence; UCMSCs-Vector and UCMSCs were defined as the negative control. Cells were cultured in 35-mm confocal dishes, respectively. When 70–80% confluence was reached, the cells were fixed and incubated with His-Tag Rabbit primary monoclonal antibody at 4℃ overnight, followed by incubation with Alexa Fluor® 594 conjugate goat anti-rabbit secondary antibodies (#8889, Cell Signaling Technology, The USA) at room temperature for 45 min. After washing three times, the nuclei were stained with DAPI staining solution for 3 min. The images were detected and captured in a single photon confocal microscopy (Ti-E A1, Nikon, Japan).

Zhang J., Zhong M., Zhong W., Lan Y., Yuan Z., Duan Y, & Wei Y. (2022). Construction of tandem diabody (IL-6/CD20)-secreting human umbilical cord mesenchymal stem cells and its experimental treatment on diffuse large B cell lymphoma. Stem Cell Research & Therapy, 13, 473.

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