Under general anesthesia (2% isoflurane), we removed the Kwik-sil and PRECLUDE® Pericardial membrane that was placed during the laminectomy procedure. Then, we immediately covered the spinal cord with a saline-soaked sponge (Ethicon, Surgifoam®). We cut a Teflon AF film (VICI Metronics, Poulsbo, WA; Teflon AF 2400, 50 μm thick) to the size of the exposed spinal cord and placed it directly over the leptomeninges. We placed Kwik-Sil above the Teflon AF then adhered a 3.0-mm glass coverslip (#0, CS-3R-0, Warner Instruments) on top, forming a multi-layered tier above the spinal cord, and we allowed it to harden for 10 minutes. We then added a supportive coat of bone cement or Norland UV-curable optical adhesive (NOA 81) from the base of the implant to the coverslip. We administered Carprofen (5.0 mg/kg, sc) for postoperative analgesia.
Cs 3r 0
Manufactured by Warner Instruments
The CS-3R-0 is a laboratory instrument designed for cell culture applications. It features three independent cell culture chambers with a controlled environment, including temperature, humidity, and gas composition regulation. The core function of the CS-3R-0 is to provide a stable and optimized environment for cell growth and experimentation.
Automatically generated - may contain errors
Lab products found in correlation
3 protocols using cs 3r 0
1
Spinal Cord Protective Implant Procedure
2
Fluorescent Bead Imaging Protocols
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Sample holders for fluorescent beads were built by gluing smaller coverslips (3 mm ⌀, CS-3R-0, Warner Instruments) to a 2-mm glass capillary tube (Harvard Apparatus) using silicone adhesive (Würth, Super RTV Silikon). Fluoresbrite YG Microspheres (0.20 µm size, Polysciences) were diluted in distilled water and pipetted on the coverslip. The capillary tube was clamped into a sample holder and inserted into the Schmidt objective. For imaging at an index of n = 1.45, the immersion chamber was filled with a fused silica matching liquid (50350, Cargille). Type A immersion oil (16482, Cargille) was used for measurements at n = 1.51. The liquids were chosen such that the fluorescent beads did not dissolve; other immersion media, such as BABB, DBE and ECI, rapidly dissolved Fluoresbrite beads. For each immersion medium and FOV location, eight beads were measured, and the resulting PSFs were fitted with Gaussian profiles in x–y–z.
3
Chronic Imaging Window Implantation in Mice
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Chronic imaging window surgery Four male GCaMP6f mice were used for all the imaging experiments in this study. Mice were anesthetized with isoflurane (1.5%-2.0%) and a craniotomy was centered at À2.0 mm AP and 1.6 mm ML from bregma over the left hemisphere using a 3 mm diameter trephine drill bit (Trephine Bur, 3.0/2.0mm, THB30, Osung USA). The cortex and corpus callosum, but not alveus, above the dorsal hippocampus was removed by gentle suction with saline using a 27 gauge blunt-tip needle. After cortex / callosum removal, a glass coverslip (T 0.1 mm, D 3 mm, round, No.0, Warner Instruments, CS-3R-0) that was previously attached (using (Norland Optical adhesive 81) to the bottom of stainless steel cannula (OD 3.02 mm, ID 2.54 mm, L 1.75 mm, New England Small Tube Corp. Type 304, stainless steel, 11TW) was implanted over the dorsal CA1 region. A thin layer of Kwik-Sil (WPI) was applied between the cannula and skull. The top part of the cannula and custom headplate (stainless or titanium) were fixed to the skull using dental cement (Meta-bond) and dental acrylic. Body temperature was maintained at 37 C using a temperature controlled heating pad.
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