Loading samples into SLICE

Overview

Cleared-tissue specimens must be immobilized inside a SLICE Cuvette, containing mounting medium and sealed with a SLICE Cuvette quick-release mount in order to image them on a SLICE system. Position and immobilize the sample inside the cuvette using a Brain Bracket, then submerge in storage solution (that is compatible with the clearing method used to prepare the sample) to maintain clarity and prevent drift during imaging. It is not strictly required to use a Brain Bracket, however, they enable convenient tissue handling and stable positioning. A thin layer of silicone sealant applied at the cuvette‑mount junction can prevent mounting medium from evaporating and is recommended for imaging and storing samples. Once assembled, secure the sealed cuvette onto the SLICE microscope stage; SLICE Cuvette quick-release mounts are stably held on the stage by magnets.

Chemical risks to people and equipment

Employ chemical-safety precautions when handling cleared tissue or tissue storage solution. Wear personal protective equipment appropriate for the chemicals present in your cleared specimen.

Chemicals used in tissue-clearing protocols may be hazardous, volatile, and corrosive. To minimize risk, we highly recommend that you take the following sealing the cuvette with chemically compatible adhesive or sealant after mounting samples.

Note that some of these chemicals may also be corrosive, presenting a hazard to SLICE hardware. Avoid spilling or dripping corrosive material on the instrument; clean up immediately if spills or drips do occur.

Required materials

  • SLICE cuvette, any size, MBF Bioscience, containing the following:

    • Fused quartz glass cuvette

    • SLICE quick-release coupling mount: the coupling mount is the lid for the cuvette that both seals it and magnetically connects the cuvette to the SLICE stage.

    • MBF Bioscience Mounting Bracket (available soon)

  • Cleared-tissue specimen, sized to fit in the cuvette. Compatible clearing methods include iDISCO, CLARITY, SHANEL, CUBIC, BINAREE and other optical clearing techniques.

  • Mounting medium appropriate for the tissue-clearing method: the sample will be submerged in mounting medium for imaging

  • Adhesive for mounting the sample and sealing it in the cuvette. We have used this clear silicone sealant with many clearing techniques, but it is crucial to use an adhesive that is chemically compatible with your clearing reagents and storage medium.

  • (optional) Thin, heat resistant, transparent, silicone sheet, cut into slices that fit into the cuvette you will be using. We have used this product.

  • Safety gear and supplies, including personal protective equipment (PPE), appropriate for the clearing method used for sample preparation and storage.

Sample Loading Procedure

Click below to view a short video of loading sample into a cuvette and mounting it onto the SLICE stage.

A. Use a Mounting Bracket to place sample in the cuvette

  1. Attach cleared tissue to an MBF Bioscience Mounting Bracket as follows:

    1. Gently remove storage solution from cleared tissue using a lint-free tissue.

    2. Apply small amount of adhesive to the bottom and back surfaces of the Mounting Bracket.

    3. Position the cleared tissue onto the adhesive in the desired orientation.

    4. Apply gentle pressure for 5 minutes to secure the tissue.

    5. Allow to dry for 1 hour until cured.

  2. Insert Mounting Bracket into cuvette and add mounting medium to cover the sample:

    1. Gently insert the Mounting Bracket into the cuvette and settle it fully at the bottom.

    2. Fill the cuvette with mounting medium until brain tissue is fully immersed.

    3. Remove any bubbles that may have formed in the cuvette using a pipette and wipe away any mounting medium on the exterior of the cuvette.

alt A. Place sample in the cuvette without using a Mounting Bracket

  1. Place the brain into the cuvette as follows:

    1. Add a small amount of mounting medium to the bottom of the cuvette.

    2. Carefully place the brain sample into the cuvette and adjust its orientation for imaging. Minimize the time brain is exposed to air so that it does not become dried out.

    3. Add more mounting medium to fully immerse the brain tissue.

  2. Add pieces of silicone sheet to immobilize the tissue and add mounting medium to cover the sample:

    1. Gently push the brain toward the side of the cuvette that will face the detection objective.

    2. Stack several silicone slices to form a block that will fill the space between the brain and the cuvette surface opposite the detection objective.

      Trim the silicone slices so that the block fits into the cuvette without scratching it.

    3. Insert the trimmed silicone block into the cuvette to hold the brain in position.

    4. Remove any bubbles that may have formed in the cuvette using a pipette and wipe away any mounting medium on the exterior of the cuvette.

B. Seal the cuvette and attach it to the SLICE stage

  1. Seal the cuvette with a SLICE quick-release coupling mount.

    1. Place the cuvette mount onto the cuvette with gentle pressure. Avoid using excessive force to prevent breaking or cracking the cuvette.

    2. Apply a thin layer of adhesive or sealant around the junction to secure the mount.

    3. Allow to dry for 1 hour until cured.

  2. Attach cuvette to the SLICE stage.

    1. Tilt the cuvette slightly and hook the right slot of the magnet mount onto the right stage rod from below.

    2. Use the right rod as a pivot and rotate the cuvette clockwise until the left slot engages with the left rod.