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Nunc lab tek 2 8 chamber slide

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The Nunc Lab-Tek II 8-Chamber Slide is a laboratory equipment designed for cell culture and microscopy applications. It consists of a glass slide with 8 individual chambers that allow for the simultaneous cultivation and observation of multiple cell samples.

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Lab products found in correlation

Immu mount, by Thermo Fisher Scientific (4 mentions) Carl lsm 710 confocal microscope, by Zeiss (2 mentions) Eclipse ti e microscope, by Nikon (1 mentions) Fluoroshield, by Merck Group (1 mentions) Dylight 488, by Vector Laboratories (1 mentions) Anti pan adp ribose binding reagent, by Merck Group (1 mentions) Ds qi1 widefield camera, by Nikon (1 mentions) Dylight 594, by Vector Laboratories (1 mentions) Viewrna cell plus assay kit, by Thermo Fisher Scientific (1 mentions) Fv1000 confocal microscope, by Olympus (1 mentions) Alexa fluor 488, by Thermo Fisher Scientific (1 mentions) Phrodo red dextran, by Thermo Fisher Scientific (1 mentions) Lyovec, by InvivoGen (1 mentions) Hoechst 33342, by Thermo Fisher Scientific (1 mentions) α tubulin, by Santa Cruz Biotechnology (1 mentions) Cdc25c, by Santa Cruz Biotechnology (1 mentions) Aurka, by Cell Signaling Technology (1 mentions) Lsm880 confocal microscope system, by Zeiss (1 mentions) Apoptag red in situ apoptosis detection kit, by Merck Group (1 mentions)

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Lab-Tek chamber slides (385 citations) Nunc Lab-Tek (110 citations) Nunc Lab-Tek II (60 citations) Lab-Tek II chamber (24 citations) Nunc™ Lab-Tek™ Chamber (20 citations) NuncTM Lab-TekTM (4 citations) Nunc Lab-Tek II chamber (3 citations) NuncTM Lab-TekTM II (3 citations) Lab-Tek II slides (3 citations) Lab Tek II 8-chamber (2 citations)

8 protocols using nunc lab tek 2 8 chamber slide

1

DNA Damage Response Quantification

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Cells were cultured directly onto Nunc Lab-Tek II 8 Chamber Slides (Thermo Scientific) and treated in serum-free medium for 1 hour, in the presence or not of MMS (1 mM). After treatment, cells were incubated in fresh complete medium for 1 hour and subsequently fixed in 4% paraformaldehyde. Fixed cells were permeabilised using a 1:1 methanol:acetone solution for 10 minutes at room temperature. Cells were then blocked for 1 hour with 10% normal goat serum (NGS) and labeled using anti-γH2AX phospho S139 (1:500, Abcam) antibody and an anti-pan-ADP-ribose binding reagent (1:1000, Millipore), at 4 °C overnight. Primary antibodies were prepared in 1% NGS to reduce background staining. Slides were washed with TBS, 0.1% Tween-20 prior to incubation with secondary antibodies. Secondary antibodies used were goat anti-rabbit DyLight 488 (Vector Laboratories) and goat anti-mouse DyLight 594 (Vector Laboratories), 1:1000 in TBS containing 1% NGS. All incubation steps were performed protected from light in a humidified chamber. Slides were mounted using Fluoroshield (Sigma). Samples were imaged using a Nikon A1M laser scanning confocal microscope and DS-Qi1 Widefield Camera (on an Eclipse Ti-E microscope). Fluorescence intensity measurements were performed using Fiji (NIH, Bethesda, MD).
Alhumaydhi F.A., de O. Lopes D., Bordin D.L., Aljohani A.S., Lloyd C.B., McNicholas M.D., Milano L., Charlier C.F., Villela I., Henriques J.A., Plant K.E., Elliott R.M, & Meira L.B. (2020). Alkyladenine DNA glycosylase deficiency uncouples alkylation-induced strand break generation from PARP-1 activation and glycolysis inhibition. Scientific Reports, 10, 2209.
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2

LINC00844 Expression Visualization in HepG2 Cells

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HepG2-LINC00844 cells were seeded on collagen-coated Nunc™ Lab-Tek™ II 8-chamber slides (ThermoFisher) and cultured to 100% confluency. RNA FISH was performed using a ViewRNA™ Cell Plus Assay kit (ThermoFisher) according to the manufacturer’s instructions. Cells were first fixed and permeabilized with the ViewRNA™ Cell Plus Fixation/Permeabilization Solution for 30 min and then fixed with Fixation Solution for 1 h at room temperature. For target probe hybridization, fixed cells were incubated in Probe Set Solution containing the LINC00844 ViewRNA cell plus probe set conjugated with Alexa Fluor 488 (ThermoFisher) for 2 h at 40 ± 1 °C. Signal amplification was then conducted by subjecting cells to sequential incubations in pre-warmed (40 ± 1 °C) PreAmplifier Solution, Working Amplifier Solution, and Working Label Probe Mix Solution for 1 h each at 40 ± 1 °C. All incubations were performed in a humidified staining tray. Chambers were removed upon completion of the hybridization protocol, and slides were mounted with VECTASHIELD® ProLong™ Gold Antifade Mountant with DAPI (Vector Laboratories, Burlingame, CA) and covered with coverslips. Fluorescent images for LINC00844 detection and DAPI (a nucleus marker) staining were taken within the following 24 h using an Olympus FV1000 confocal microscope (Olympus, Center Valley, PA).
Li D., Wu L., Knox B., Chen S., Tolleson W.H., Liu F., Yu D., Guo L., Tong W, & Ning B. (2020). Long noncoding RNA LINC00844-mediated molecular network regulates expression of drug metabolizing enzymes and nuclear receptors in human liver cells. Archives of toxicology, 94(5), 1637-1653.
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3

Microglia and Neuron Uptake of miRNAs

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60,000 microglia or 250,000 neurons isolated from C57BL/6 mice were plated per well of a Nunc™ Lab-Tek™ II 8 Chamber Slide™ (Thermo Fischer Scientific, #154534PK, Waltham, MA, USA) and were incubated with 40 μg/ml pHrodo™ Red Dextran (Thermo Fischer Scientific, #P10361, Waltham, MA, USA) for 20 min at 37 °C. Following washing steps, 5 μg/ml fluorescence-labeled miRNAs (Alexa488-miR-298-5p, Alexa488-miR-100-5p or Alexa488-let-7b-5p) complexed to LyoVec (InvivoGen #LYEC-RNA, San Diego, CA, USA) per well were added. After 4 h, cells were washed, stained with DAPI, fixed with 4% PFA, and analyzed by an SP8 confocal laser microscope with sequential analysis (Leica Biosystems, Nussloch, Germany).
Wallach T., Mossmann Z.J., Szczepek M., Wetzel M., Machado R., Raden M., Miladi M., Kleinau G., Krüger C., Dembny P., Adler D., Zhai Y., Kumbol V., Dzaye O., Schüler J., Futschik M., Backofen R., Scheerer P, & Lehnardt S. (2021). MicroRNA-100-5p and microRNA-298-5p released from apoptotic cortical neurons are endogenous Toll-like receptor 7/8 ligands that contribute to neurodegeneration. Molecular Neurodegeneration, 16, 80.
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4

Filipin Staining of Cholesterol in HUVEC

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Filipin staining was performed as descried with slight modifications [17 (link)]. HUVEC were cultured in a Nunc Lab-Tek II 8-Chamber Slide (Thermo Scientific, Rockford, IL) at 1 × 104cells/well. Cells were treated with SERM with or without cholesterol and cyclodextrin complex for 24 h. Cells were then fixed with 4% paraformaldehyde for 20 min at room temperature and stained with filipin at a final concentration of 50 μg/ml in the dark for 1 h at room temperature. Cells were washed with PBS, mounted with Immu-mount (Thermo Scientific), and observed under a Zeiss 510 Meta multiphoton confocal microscope (Carl Zeiss, Thornwood, NY).
Shim J.S., Li R.J., Lv J., Head S., Yang E.J, & Liu J.O. (2015). Inhibition of angiogenesis by selective estrogen receptor modulators through blockade of cholesterol trafficking rather than estrogen receptor antagonism. Cancer letters, 362(1), 106-115.
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5

Immunostaining and Filipin Staining of HUVEC

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HUVEC were cultured in a Nunc Lab-Tek II 8-Chamber Slide (Thermo Scientific, Rockford, IL) at 2 × 104 cells/well and treated with compounds for indicated time. Cells were then fixed with 4% paraformaldehyde for 20 min at room temperature. For general immunostaining of proteins, cells were washed by PBS and permeabilized with 0.5% Triton X-100 or 0.2% saponin (for co-staining with filipin) for 20 min. Cells were blocked with 3% bovine serum albumin (BSA) in PBS containing 0.1% Tween-20 for 1 h and incubated with primary antibodies overnight at 4ºC. Cells were then incubated with secondary antibodies conjugated with Alexa Fluor 488 or Alexa Fluor 647 for 1 h at room temperature. The cellular nuclei were stained with Hoechst 33342. Cells were rinsed with PBS, mounted with Immu-mount (Thermo Fisher Scientific), and observed under a Carl Zeiss LSM 710 confocal microscope (Carl Zeiss, Thornwood, NY). For filipin staining of cholesterol, cells were fixed with 4% paraformaldehyde for 20 min, washed with PBS, and stained with filipin at a final concentration of 50 µg/ml for 1 h at room temperature in a darkroom. Cells were then washed with PBS and mounted with Immu-mount prior to observation under the confocal microscope.
Lyu J., Yang E.J., Head S.A., Ai N., Zhang B., Wu C., Li R.J., Liu Y., Chakravarty H., Zhang S., Tam K.Y., Dang Y., Kwon H.J., Ge W., Liu J.O, & Shim J.S. (2018). Astemizole Inhibits mTOR Signaling and Angiogenesis by Blocking Cholesterol Trafficking. International Journal of Biological Sciences, 14(10), 1175-1185.
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6

Immunofluorescence Analysis of Cell Cycle Regulators

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Cells were seeded on a Nunc Lab-Tek II 8-Chamber Slide (Thermo Fisher Scientific) and treated with compound or siRNA for the indicated time. Cells were then fixed with 4% paraformaldehyde and permeabilized with 0.5% Triton X-100, followed by blocking with 2% bovine serum albumin for 1 h. Cells were incubated with primary antibodies, including AURKA (Cell Signaling, #14475s, 1:100 dilution), CDC25C (Santa Cruz, sc-327, 1:100 dilution), and α-tubulin (Santa Cruz, sc-5286, 1:100 dilution) in the blocking buffer overnight at 4 °C, followed by the incubation with secondary antibodies conjugated with Alexa Fluor 488 or Alexa Fluor 647 for 1 h at room temperature. The nuclei were stained with Hoechst 33342 (Thermo Fisher Scientific). After washing with phosphate-buffered saline (PBS), cells were mounted with Immu-mount (Thermo Fisher Scientific) and observed under a Carl Zeiss LSM 710 confocal microscope (Carl Zeiss, Thornwood, NY).
Wu C., Lyu J., Yang E.J., Liu Y., Zhang B, & Shim J.S. (2018). Targeting AURKA-CDC25C axis to induce synthetic lethality in ARID1A-deficient colorectal cancer cells. Nature Communications, 9, 3212.
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7

Immunofluorescence Staining of Cells

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Cells were seeded into a Nunc Lab-Tek II 8-Chamber Slide (Thermo Fisher Scientific). After treatment, cells were fixed with 4% paraformaldehyde at room temperature for 30 min, and then blocked with 3% BSA in PBS for 30 min. Cells were then incubated with primary antibodies at 4 °C overnight and incubated with secondary antibodies conjugated with Alexa Fluor 488 for 1 h at room temperature in the dark. The cellular nuclei were stained with 1 μg/mL Hoechst 33342 for 10 min. Cells were mounted with Immu-mount (Thermo Fisher Scientific) and observed under a Zeiss LSM 880 Confocal microscope System (Carl Zeiss).
Ren G., Yang E.J., Tao S., Mou P.K., Pu Y., Chen L.J, & Shim J.S. (2023). MDM2 inhibition is synthetic lethal with PTEN loss in colorectal cancer cells via the p53-dependent mechanism. International Journal of Biological Sciences, 19(11), 3544-3557.
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8

Quantifying Apoptosis in Macrophages Using TUNEL

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To assay for apoptosis using the TUNEL assay, 4 × 104 RAW264.7 cells or primary macrophages in SFM were seeded into the wells of a Nunc Lab-TekII 8-chamber slide (ThermoFisher). The slides were incubated at 37 °C in 5% CO2 for 18 h and washed twice with SFM. Into each well was added 200 µL of SFM containing 5 µg of either normal-eyecup or EAU-eyecup ESMs. The cultures were incubated for another 18 h, the supernatant was removed, and the cells were fixed in 4% paraformaldehyde (Electron Microscope Sciences, Hatfield, PA, USA) for 10 min. The cells were stained using ApopTag Red In Situ Apoptosis Detection Kit (Millipore Corp, Temecula, CA, USA), and counterstained with DAPI (MPI Biomedical, Irvine, CA). After staining, the cells were imaged by fluorescence microscopy using a FSX100 inverted fluorescent imaging microscope (Olympus, Center Valley, PA, USA).
Sanjiv N., Osathanugrah P., Fraser E., Ng T.F, & Taylor A.W. (2021). Extracellular Soluble Membranes from Retinal Pigment Epithelial Cells Mediate Apoptosis in Macrophages. Cells, 10(5), 1193.
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