Vt1200s slicer

Manufactured by Leica

The VT1200S is a versatile slicer designed for high-quality tissue sectioning. It features a vibrating blade that allows for precise and consistent cutting of samples, making it suitable for a variety of tissue types and research applications.

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3 protocols using vt1200s slicer

Accessing Bipolar Cells in Mouse Retina

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4 to 5 weeks old mice were injected with 1 or 1.5 μL volume of AAV2-7m8 carrying CoChR (∼10¹⁰ vg) under a promoter specific for RBCs(In4s-In3-200En-mGluR500P) (Lu et al., 2016 (link)). 4 to 10 weeks after the injection, the animals were anesthetized with isoflurane and killed by cervical dislocation. Eyeballs were enucleated and dissected under white light. To have a better access to the BCs with the patch pipette, we removed the photoreceptor layer using a vibratome (Leica VT1200S slicer). This procedure was previously described in details (Clérin et al., 2014 (link)). Briefly, the dissected retina was transferred in the vibratome tank filled with bubbled Ames. The retina was placed photoreceptors down on a gelatin block in the center of the tank and the solution was removed to permit the sealing of the flat-mounted retina. Once the retina was sealed, the tank was filled with bubbling Ames again and the vibratome’s blade was lowered until the GCs level. A slice of ∼80–90 μm was cut and transferred to the recording chamber under the microscope with GCs down. BCs were thus on the upper side without the photoreceptors on top of them, which made them more accessible to patch recordings (Figures 2B and 2C).

Spampinato G.L., Ronzitti E., Zampini V., Ferrari U., Trapani F., Khabou H., Agraval A., Dalkara D., Picaud S., Papagiakoumou E., Marre O, & Emiliani V. (2022). All-optical inter-layers functional connectivity investigation in the mouse retina. Cell Reports Methods, 2(8), 100268.

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Hippocampal Slice Preparation and Recording

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Three weeks after the tamoxifen induction, adult mice (Nestin-creER^T2; tdTOM) were deeply anesthetized and transcardially perfused with chilled (4 °C) dentate gyrus dissection buffer containing (in mM): 110 Choline Cl, 2.5 KCl, 1.3 KH₂PO₄, 25 NaHCO₃, 10 D-glucose, 0.6 Na Pyruvate, 1.3 Na Ascorbate at pH 7.4, 300 mOsm, and aerated with 95% O2/5% CO2, including 0.5 CaCl2, 7 MgCl2, and 5 Kynurenate acid. Transverse hippocampal slices (250 µm thickness) were obtained on a Leica VT1200S slicer, and then recovered for 30 min at 35 °C and for another 15 min at room temperature in artificial cerebrospinal fluid (ACSF) containing (in mM): 125 NaCl, 2.5 KCl, 1.3 KH₂PO₄, 25 NaHCO₃, 1.3 Na Pyruvate, 10 d-glucose, 1.3 Na Ascorbate, 2 CaCl₂, 1.3 MgCl₂ aerated with 95% O2/5% CO₂ to pH 7.4. A single slice was then transferred to a submersion chamber and perfused at 3 ml/min with aerated ACSF at 30 °C ready to be recorded.

Ma Z., Zang T., Birnbaum S.G., Wang Z., Johnson J.E., Zhang C.L, & Parada L.F. (2017). TrkB dependent adult hippocampal progenitor differentiation mediates sustained ketamine antidepressant response. Nature Communications, 8, 1668.

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Preparation of Mouse Brain Slices

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Three hundred micrometer thick parasagittal slices, were prepared 4–10 weeks after viral injection, in accordance with European guidelines. Mice were decapitated. Slices were prepared using a Leica VT1200S slicer. The sucrose slicing solution contained (in millimoles of compound used to make a liter of solution): 25 glucose, 85 NaCl, 65 sucrose, 0.5 CaCl₂, 4 MgCl₂, 2.5 KCl, 1.25 NaH₂PO₄ and 26 NaHCO₃. It was saturated with 95% O₂/5% CO₂. Slices were then incubated at 32–33°C for 30 min in a sucrose recovery solution containing (in millimoles of compound used to make a liter of solution): 25 glucose, 115 sucrose, 1 CaCl₂, 2.5 MgCl₂, 105 NaCl, 2.5 KCl, 1.25 NaH₂PO₄ and 26 NaHCO₃ saturated with 95% O₂/5% CO₂.

Chaigneau E., Ronzitti E., Gajowa M.A., Soler-Llavina G.J., Tanese D., Brureau A.Y., Papagiakoumou E., Zeng H, & Emiliani V. (2016). Two-Photon Holographic Stimulation of ReaChR. Frontiers in Cellular Neuroscience, 10, 234.

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