Ts100 fluorescence microscope

Manufactured by Nikon

Sourced in Japan

The TS100 is a fluorescence microscope designed for laboratory use. It is capable of visualizing and analyzing fluorescently labeled samples. The core function of this device is to provide high-quality imaging of fluorescent specimens.

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Lab products found in correlation

Glass bottom chambers, by MatTek (1 mentions) Leica tcs sp5 2 confocal microscope, by Leica Microsystems (1 mentions) Facscalibur flow cytometer, by BD (1 mentions) 4 6 diamidino 2 phenylindole hydrochloride dapi, by Thermo Fisher Scientific (1 mentions) Fluorescein isothiocyanate conjugated phalloidin, by Thermo Fisher Scientific (1 mentions) Hoechst, by Thermo Fisher Scientific (1 mentions) Fel0500, by Thorlabs (1 mentions) Mitomycin, by Merck Group (1 mentions)

8 protocols using ts100 fluorescence microscope

Visualizing VSV-GFP Infection and Cellular Uptake

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The VSV-GFP is a recombinant virus expressing the green fluorescent protein. The Cy3 fluorescein was modified on the RVG to provide a red fluorescent signal. In the cellular model, the virus-infected N2a cells (MOI of 1) were incubated with 100 μL of T-705@MSN-RVG (2 mg/mL) at 37 °C for 1h. Then, cells were washed three times and stained by a Hoechst (Thermo Fisher, Waltham, MA, USA) at 37 °C for 5 min. Subsequently, the cells were visualized by a TS100 fluorescence microscope (Nikon, Tokyo, Japan).
In the mouse infection model, co-localization was determined in the mouse brain. Briefly, six-week-old female Balb/c mice were infected with VSV-GFP at 200 FFU or DMEM by intracranial (i.c.) route. Then, the mice were inoculated with 25 μL of T-705@MSN-RVG (1 mg/mL) by i.v. injection. The mouse brains were collected at 2 dpi and fixed in 4% paraformaldehyde for 48 h, followed by being dehydrated in 30% sucrose solution and embedded in SAKURA Tissue-Tek^® O.C.T compound (SAKURA, Torrance, CA, USA) for rapid freezing and slicing [35 (link)]. The slices were stained by Hoechst (Thermo Fisher, Waltham, MA, USA) at 37 °C for 5 min and then visualized by a TS100 fluorescence microscope (Nikon, Tokyo, Japan).

Ren M., Zhou Y., Tu T., Jiang D., Pang M., Li Y., Luo Y., Yao X., Yang Z, & Wang Y. (2023). RVG Peptide-Functionalized Favipiravir Nanoparticle Delivery System Facilitates Antiviral Therapy of Neurotropic Virus Infection in a Mouse Model. International Journal of Molecular Sciences, 24(6), 5851.

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Fluorescence and Confocal Microscopy Analyses

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Fluorescence and light microscopy analyses were performed by a Nikon TS100 fluorescence microscope (Nikon Corporation) and a Leica TCS SP5 II confocal microscope (Leica Microsystem) with 488, 543 and 633 nm illumination and oil-immersed objectives. For confocal live imaging, cells were grown on MatTek glass bottom chambers (MatTek Corporation). The images were further processed and analyzed for mean fluorescence quantification in ImageJ software (National Institutes of Health), using the JACoP plug-in for colocalization studies [76 (link)]. We used the Pearson's coefficient as the parameter to measure colocalization in our samples.

Canonico B., Cesarini E., Salucci S., Luchetti F., Falcieri E., Di Sario G., Palma F, & Papa S. (2016). Defective Autophagy, Mitochondrial Clearance and Lipophagy in Niemann-Pick Type B Lymphocytes. PLoS ONE, 11(10), e0165780.

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Nucleofection of Adipose-Derived Stem Cells

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The nucleofection of the ADSCs was performed according to the optimized protocols provided by the manufacturer (Amaxa Biosystems, Cologne, Germany). Briefly, prior to nucleofection, a Petri dish culture containing 1 ml of DMEM was incubated in the CO₂ incubator at 37°C. The ADSCs were digested and centrifuged following the adjustment of the density of the suspended cells to 2×10⁶/ml. The cells (5×10⁵) and plasmid (2 µg) were suspended in 100 µl prewarmed nucleofector solution (Amaxa Biosystems). For nucleofection, the program U-23 was selected. Immediately, following nucleofection, the cells were transferred into prewarmed fresh medium in six-well plates, and incubated in a CO₂ incubator at 37°C and monitored daily. The cells were analyzed 24 h post-nucleofection for transfection efficiency by FACS (FACSCalibur flow cytometer; Becton-Dickinson) and a Nikon Ts100 fluorescence microscope (Nikon Corp., Tokyo, Japan) for GFP expression. The ADSCs transduced with pcDNA3.1/OX40Ig or pcDNA3.1/GFP are referred to as ADSCs^OX40Ig or ADSCs^GFP, respectively.

Liu T., Zhang Y., Shen Z., Zou X., Chen X., Chen L, & Wang Y. (2016). Immunomodulatory effects of OX40Ig gene-modified adipose tissue-derived mesenchymal stem cells on rat kidney transplantation. International Journal of Molecular Medicine, 39(1), 144-152.

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Viral Infection Time-Course Quantification

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For viral infection, the N2a cells were infected with VSV and RABV at a multiplicity of infection (MOI) of 0.01 or 1 for 1 h at 37 °C. Subsequently, the cells were washed three times with PBS and maintained in DMEM supplemented with 2% FBS for 12, 24, 36, and 48 h at 37 °C. The virus titration was performed according to the previous study [34 (link)]. Briefly, the viral supernatant was diluted in a 10-fold gradient (from 10⁻¹ to 10⁻⁷ dilutions) to be added in quadruplicate to 96-well cell culture plates at 100 μL, followed by supplying 100 μL BSR cells (1 × 10⁴ cells/mL) in each well. After 48 h of incubation, the cells were visualized by a TS100 fluorescence microscope (Nikon, Tokyo, JPN).

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Immunohistochemistry of FFPE Tissue Sections

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Formalin-fixed, paraffin-embedded (FFPE) tissue blocks were sectioned at 5 μm, baked for 1.5 h for deparaffinization at 65°C, and rehydrated prior to antigen retrieval using a standard xylene/alcohol protocol. Endogenous peroxidase activity was blocked with 3% hydrogen peroxide in TBST 5 min. An antigen retrieval step in citrate buffer (10 mM, pH 6) for 10 min under low microwave power was conducted. The blocker was drained, and gremlin1 and BMP-7 (Santa Cruz, USA) primary antibodies were applied at a concentration of 1 μg/mL. Control sections were incubated with the appropriate IgG negative control. Slides were incubated at 37°C for 1 h, followed by overnight incubation at 4°C with primary antibodies at 1:50 dilution, and subsequently for 1 h at 37°C with FITC-labeled rabbit anti-goat IgG (H+L) (NovoGene Biotech, Wuhan, China). Sections were contrasted and imaged under a TS100 fluorescence microscope (Nikon, Tokyo, Japan).

Zeng X.Y., Zhang Y.Q., He X.M., Wan L.Y., Wang H., Ni Y.R., Wang J., Wu J.F, & Liu C.B. (2016). Suppression of hepatic stellate cell activation through downregulation of gremlin1 expression by the miR-23b/27b cluster. Oncotarget, 7(52), 86198-86210.

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Cytocompatibility Assessment of Ternary Composite Mat

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The cytocompatibility of the ternary composite mat was assessed via MTT assay. In brief, coverslips and scaffolds were fixed in 24-well plates with stainless-steel rings and sterilized by exposure to 75% alcohol solution for 2 hrs. After that, all of the samples were washed three times with PBS solution and soaked in DMEM overnight before cell seeding. Then, mouse fibroblast cells (L929) were seeded at a density of 1.5 × 10⁴ cells per well for 2, 4, 8, and 12 hrs for cell adhesion assay and 1, 3, 5, and 7 d for cell proliferation assay. Coverslips without scaffolds were used as controls.
MTT assay and SEM observation were employed to separately evaluate the viability and morphology of the adhered and proliferated L929 cells cultured onto different scaffolds (n=3 for each group) according to the manufacturer’s protocol of our previous research.26 (link) On a separate set of sheets, the attached cells were fixed with 4% paraformaldehyde and then 4′,6′-diamidino-2-phenylindole hydrochloride (DAPI, Invitrogen, USA) and fluorescein isothiocyanate-conjugated phalloidin (Invitrogen, USA) were used to stain the nucleus and cytoskeletons of cells. Specimens were observed under a TS100 fluorescence microscope (Nikon, Japan).

Zhu T., Jiang J., Zhao J., Chen S, & Yan X. (2019). Regulating Preparation Of Functional Alginate-Chitosan Three-Dimensional Scaffold For Skin Tissue Engineering. International Journal of Nanomedicine, 14, 8891-8903.

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Portable Fluorescence Microscopy Protocol

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Benchtop microscopic imaging was accomplished by placing the cell capture interface (layer 2) on a glass slide and imaged using a 10X objective and a filter cube for blue bandpass filter excitation and green long-pass filter emission. Nikon TS100 fluorescence microscope (Tokyo, Japan) was used for imaging.
Smartphone-based microscopic imaging was accomplished using a smartphone (iPhone 8; Apple Inc., Cupertino, CA, USA) connected to a commercially available microscope attachment (XFox Professional 300X Optical Glass Lenses; X&Y Ind., Shenzhen, China), attached to a 500 nm long-pass optical filter (FEL0500, Thorlabs Inc., Newton, NJ, USA). Again, smartphone imaging was also accomplished by sandwiching the WBC capture interface between the optical filter and a transparent glass surface. During detection the WBC capture interface is illuminated using a 466 nm LED (TLHB5100; Vishay, Malvern, PA, USA) powered by a 9 V battery. The current is limited using a 500 Ω resistor.

Bills M.V., Nguyen B.T, & Yoon J.Y. (2019). Simplified White Blood Cell Differential: An Inexpensive, Smartphone- and Paper-Based Blood Cell Count. IEEE sensors journal, 19(18), 7822-7828.

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Wound Healing Assay for U-87 and HF Cells

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U-87 cells and HF were seeded at a density of 3 × 10⁵ cells in a 6-well plate in complete culture media and grown to confluence. The day after, cells were treated with 4 μg/mL of mitomycin (Sigma-Aldrich) for 2 h to inhibit cell proliferation and then a wound was inflicted using a tip. After washing with phosphate-buffered saline (PBS), cells were incubated as above mentioned in comparison to untreated cells. The scratch wound was observed and photographed at different time points using an inverted-phase-contrast microscope (TS100 fluorescence microscope and video camera, Nikon, Tokyo, Japan). Three measurements per scratch were performed (two replicates/condition, experiments performed in duplicate).

Nigro E., Crescente G., Formato M., Pecoraro M.T., Mallardo M., Piccolella S., Daniele A, & Pacifico S. (2020). Hempseed Lignanamides Rich-Fraction: Chemical Investigation and Cytotoxicity towards U-87 Glioblastoma Cells. Molecules, 25(5), 1049.

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