μ slide 6 0.4 channel slides

Manufactured by Ibidi

The μ-Slide VI 0.4 channel slides are a specialized lab equipment designed for cell culture experiments. These slides feature six individual channels with a height of 0.4 mm, allowing for the efficient cultivation and observation of cells in a controlled environment.

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Lab products found in correlation

Cellevent caspase 3 7 green detection reagent, by Thermo Fisher Scientific (3 mentions) X vivo 10, by Lonza (1 mentions) Lysobritenir, by AAT Bioquest (1 mentions) Sensoplate 24 well glass bottom plates, by Greiner (1 mentions) Eclipse ti e inverted microscope, by Nikon (1 mentions) Recombinant human stem cell factor, by R&D Systems (1 mentions) Penicillin, by Thermo Fisher Scientific (1 mentions) Streptomycin, by Thermo Fisher Scientific (1 mentions) L glutamax, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

μ‐Slide VI 0.4 channels µ-Slide VI 0.4 slides μ-Slides VI μ-Slide VI μ-slide IV 6-channel µ-Slides µ-Slide VI Ibidi slides µ-Slide VI 0.4 µ-Slides VI 0.4 μ-slides

5 protocols using μ slide 6 0.4 channel slides

Live Cell Imaging of Hematopoietic Cells

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Time-lapse experiments were conducted at 37°C, 5% O₂, 5%CO₂ on μ-slide VI^0.4 channel slides (IBIDI) coated with 20 μg/ml anti-human CD43-biotin antibody(51 (link)). BFP+ cells 3 day post-lentiviral transduction were sorted and cultured overnight in phenol-red free X-VIVO 10 (Lonza) medium supplemented with BSA (1%), L-glutamine, Pen/Strep, human LDL and the cytokine cocktail described above before imaging. Brightfield images were acquired every 15 minutes for 4 days using a Nikon-Ti Eclipse equipped with a Hamamatsu Orca Flash 4.0 camera and a 10x CFI Plan Apochromat λ objective (NA 0.45). Single-cell tracking and fate assignment were performed using self-written software as previously described(51 (link)). Time to division was calculated using R 3.6.1.

Xie S.Z., Kaufmann K.B., Wang W., Chan-Seng-Yue M., Gan O.I., Laurenti E., Garcia-Prat L., Takayanagi S.I., Ng S.W., Xu C., Zeng A.G., Jin L., McLeod J., Wagenblast E., Mitchell A., Kennedy J.A., Liu Q., Boutzen H., Kleinau M., Jargstorf J., Holmes G., Zhang Y., Voisin V., Bader G.D., Wang J.C., Hannun Y.A., Luberto C., Schroeder T., Minden M.D, & Dick J.E. (2020). Sphingosine-1-Phosphate Receptor 3 Potentiates Inflammatory Programs in Normal and Leukemia Stem Cells to Promote Differentiation. Blood Cancer Discovery, 2(1), 32-53.

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Time-lapse imaging of VENUS and LysoBrite

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We performed time-lapse acquisition with YouScope (http://langmo.github.io/youscope/) on Nikon-Ti Eclipse microscopes as described previously¹² (link) with yellow fluorescent protein (500/20; 515LP; 535/30) and Cy5 (620/60; 660LP; 700/75; all, AHF) filter cubes to detect VENUS and LysobriteNIR, respectively. PCNAVENUS experiments were imaged every 20 minutes and trackSeq experiments every 30 minutes with 20× (NA 0.75) and 10× (NA 0.45) CFI Plan Apochromat λ objectives, respectively. Cells were cultured in Iscove modified Dulbecco medium¹² (link) containing 16.6 μM LysobriteNIR (22641, AAT Bioquest), at 37°C, 5% oxygen, 5% carbon dioxide in SensoPlate 24 Well Glass bottom plates (Greiner Bio-One) for trackSeq experiments or μ-slide VI^0,4 channel slides (IBIDI), coated with 2.5 or 10 μg/mL anti–CD43-biotin (eBIoR2/R60; eBioscience)¹⁴ (link) in phosphate-buffered saline at room temperature for 1 hour. The trackSeq experiments were analyzed by self-written automated live event recognition in time-lapse data (alerT) software, and others were tracked as described previously.^6,15,17 (link)

Wehling A., Loeffler D., Zhang Y., Kull T., Donato C., Szczerba B., Camargo Ortega G., Lee M., Moor A., Göttgens B., Aceto N, & Schroeder T. (2022). Combining single-cell tracking and omics improves blood stem cell fate regulator identification. Blood, 140(13), 1482-1495.

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Real-Time Imaging of CD8+ T Cell-Mediated Killing of Irradiated Tumor Cells

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Cancer cells were tagged with mCherry via lentivirus transfection (vector: pCDH-CMV-MCS-EF1α-Hygro, System Biosciences), followed by FACS to isolate the bright population. These cells were irradiated with 10 Gy IR, immediately seeded in μ-Slide VI 0.4 channel slides (ibidi), and returned to the incubator. Forty-eight hours later, CD8⁺ T cells isolated from WT or Ifnar1-KO MC38 tumors by FACS on day 4 after 10 Gy IR, labeled with CMAC dye (Thermo Fisher Scientific), were added to channel slides at an effector/tumor cell (E/T) ratio of 3:2. Cancer cells were evaluated by counting cells seeded in parallel plates to take account of their altered proliferation after IR. Culture media were supplemented with IL-2 (30 U/mL) and CellEvent Caspase-3/7 Green Detection Reagent (500 nM, Thermo Fisher Scientific). Subsequently, channel slides were imaged every 15 minutes for up to 16 hours using a Nikon ECLIPSE Ti-E Inverted Microscope. During the whole course of imaging, chamber slides were maintained at 37°C with 5% CO₂. The imaging data were transferred to an Imaris station for analysis (Bitplane). The percentage of caspase-3/7⁺ tumor cells was calculated using the following formula: number of tumor cells becoming caspase-3/7⁺ following interaction with CD8⁺ T cells/number of tumor cells interacted with CD8⁺ T cells × 100.

Chen J., Cao Y., Markelc B., Kaeppler J., Vermeer J.A, & Muschel R.J. (2019). Type I IFN protects cancer cells from CD8+ T cell–mediated cytotoxicity after radiation. The Journal of Clinical Investigation, 129(10), 4224-4238.

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Cytoplasmic and Nuclear GEM Imaging

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For cytoplasmic 40 nm GEMs, PfV encapsulin-mSapphire was expressed in fission yeast cells carrying the multicopy thiamine-regulated plasmid pREP41X-PfV-mSapphire. For nuclear 40 nm GEMs, NLS-PfV-mSapphire was inserted pREP41X. The expression of these constructs was under the control of the thiamine repressible nmt41 promoter (Maundrell 1990) . Cells were grown using a protocol that produced appropriate, reproducible expression levels of the GEMs: cells carrying these plasmids were grown from a frozen stock on EMM3S -LEU plates without thiamine for 2-3 days at 30 °C and stored at room temperature for 1-2 days to induce expression. They were then inoculated in liquid EMM3S -LEU with 0.1 μg/mL of thiamine (#T4625-25G, Sigma Aldrich) for partial repression of the nmt41 promoter and grown for one day at 30 °C to exponential phase. Cells were immobilized in lectin-treated μ-Slide VI 0.4 channel slides (#80606, Ibidi) and imaged in fields of 250×250 pixels or smaller using HILO TIRF illumination at 100 Hz for 10 s.

Shu T., Shu T., Rao G., Xie Y., Herzog N.L., Bazley A., Bonucci M., Keegan S., Saxena S., Ettefa F., Brittingham G., Lemière J., Fenyö D., Chang F., Delarue M., & Holt L.J. (2021). nucGEMs probe the biophysical properties of the nucleoplasm.

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Time-lapse Imaging of Hematopoietic Cells

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Time-lapse experiments were conducted at 37 o C, 5% O 2 , 5%CO 2 on μ-slide VI 0.4 channel slides (IBIDI) coated with 20 μg/ml anti-human CD43-biotin antibody (37) . BFP + cells were sorted 3 days post-lentiviral transduction and cultured overnight in phenol red free IMDM supplemented with 20% BIT (Stemcell Technologies), 100 ng/mL human recombinant Stem Cell Factor (SCF), 50 ng/mL human recombinant Thrombopoietin (TPO), 100 ng/mL human Fms-related tyrosine kinase 3 ligand (Flt3L, all R&D Systems), 2 mM L-GlutaMAX (Gibco), 100 U/mL penicillin and 100 µg/mL Streptomycin (Gibco) before imaging. Brightfield images were acquired every 8 minutes for 3-4 days using a Nikon-Ti Eclipse equipped with a Hamamatsu Orca Flash 4.0 camera and a 10x CFI Plan Apochromat λ objective (NA 0.45). Single-cell tracking and fate assignment were performed using self-written software as previously described (11, 35, 36) . Time to division was calculated using R 3.5.3.

García‐Prat L., Kaufmann K.B., Schneiter F., Voisin V., Murison A., Chen J., Chan‐Seng‐Yue M., Gan O.I., McLeod J., Smith S., Shoong M., PARIS D.S., Pan K., Zeng A.G., Krivdova G., Gupta K., Takayanagi S., Wagenblast E., Wang W., Lupien M., Schroeder T., Xie S.Z., & Dick J.E. (2021). Dichotomous regulation of lysosomes by MYC and TFEB controls hematopoietic stem cell fate.

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