Protocol:

Protocol for the Removal of Alvetex® Discs from 6 well or 12 Well Inserts by Unclipping the Well Insert Base

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Note: This technique works with both wet and dry Alvetex membranes and works with both Alvetex 6-well inserts (AVP004) and 12-well inserts (AVP005).

1. Equipment

  • A sterile surface on which to stand the well insert — if aseptic technique is required. A new Petri dish or lid is ideal.
  • A Nuffield spatula (see Figure 1). Alternative types of spatula/device also work: i.e. any piece of (sterilizable) metal or plastic with a curved profile, that is thin enough to unclip the well insert base hooks (see Procedure, step 3).
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Figure 1. A Nuffield type spatula

  • Tweezers — preferably with blunt, smooth gripper-pads — for handling the Alvetex membrane.
  • 70% ethanol — for sterilization of the spatula and tweezers (and the support surface).
  • Microbiological safety cabinet — if aseptic technique is required.
  • Personal protective equipment — lab coat, gloves, etc.

2. Procedure

  1. Place the well insert on a clean, sterile surface.
  2. Hold the well insert by placing two fingers under two wings (see Figure 2).
  3. Hold the spatula backwards with its underside toward the Alvetex well insert. With the spatula at an angle approximately 45° place the spatula into the well insert window so that the tip of the spatula can press behind the well insert base hook (see Figures 3 and 4)

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Figure 2. Holding a well insert on a sterile surface.

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Figure 3. Pressing the spatula tip behind the well insert base hook.

Note: in the Figure 3 photo, the plastic sides of the well insert window can be seen reflected in the shiny metal spatula underside.

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Figure 4. Drawing of a 12 well insert (AVP004) showing the location of a well insert window and a well insert base hook.

  1. While pressing down behind the hook with the spatula, use the fingers of your other hand to carefully lift the well insert upper part (see Figure 5).
  2. Meanwhile, bring the spatula to a vertical position so that its tip releases the hook on the well insert base.
  3. The well insert base may now have become completely detached from the well insert upper part.
  4. If the well insert base is still partially attached, then rotate the insert and perform steps 2-5 for un-hooking the base at another window.
  5. The Alvetex membrane can then be easily lifted out of the well insert base using tweezers (see Figure 6).

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Figure 5. Un-hooking the well insert base.

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Figure 6. Alvetex membrane removed.

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