Protocol: Evaluating Cell Viability Using Invitrogen LIVE/DEAD Cell Imaging Kit with P3D Scaffolds

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Application

This protocol explains how to perform Cell viability using Invitrogen LIVE/DEAD Cell Imaging Kit with Ossiform P3D Scaffolds.

Materials

  • Invitrogen LIVE/DEAD cell imaging kit
  • Fresh media
  • Microscope with FTIC/GFP and Texas Red filter
  • Tinfoil for covering the samples during incubation time

Flowchart

FIGURE 1: WORKFLOW FOR EVALUATING CELL VIABILITY USING THE LIVE/DEAD CELL IMAGING KIT BY INVITROGEN.

Figure 1.: Workflow for evaluating cell viability using the LIVE/DEAD cell imaging kit by Invitrogen.

Notes before starting and general advice on material handling

  • All handling of the P3D Scaffolds products should be performed using gloves, according to the standard aseptic methods.
  • The scaffolds are supplied sterile by dry heat sterilization and remains sterile until opened.

Procedure

  1. Thaw the cell kit at the room temperature.
  2. Aspirate the media from the scaffold.
  3. Add 200 μL fresh media to each scaffold.
  4. Mix the Live/Dead stain master mix by adding 1 mL green staining solution to the 1 μL red staining solution – enough for five Ø12mm P3D Scaffold samples. Please upscale the master mix according to your experimental setup.
  5. Add 200 μL from the Live/Death cell kit solution to each scaffold.
  6. Cover with tinfoil paper to prevent it from the light damage.
  7. Incubate for 15 min at room temperature.
  8. Make the analyses by using fluorescent microscope:
    • For the live cells, use FTIC or GFP filters.
    • For the dead cells, use the Texas Red filters.

Disclaimer

The products are “For Research Use Only (RUO)” and should not be used for clinical, diagnostic, or therapeutic purposes. Ossiform and its distributor, Ilex Life Sciences LLC, make no other warranties, expressed or implied, including the implied warranty of merchantability and the implied warranty of fitness for particular purpose.

Acknowledgement

This protocol has been adapted from the protocol published by Ossiform. Ilex Life Sciences is an authorized distributor of Ossiform P3D Scaffolds research product line.

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