Product FAQ

Which cell viability kits are compatible with fixation?

Answer

The LIVE/DEAD Fixable kits for flow cytometry analysis are compatible with fixation. These kits use amine-reactive cell-impermeant dyes that stain the cell surface of live cells and also the cytosol of dead cells-live cells are dim and dead cells are bright. Since the dye is covalently bound to the cells, it will be retained after fixation. Unfortunately, this method does not work well for imaging-based assays, as all cells are stained and it is difficult to distinguish bright dead cells from dim live cells with a microscope. Ethidium monoazide (EMA; Cat No. E1374) is a cell impermeant nucleic acid stain that can be applied to live cultures and stains only dead cells. After incubation and washing away unbound dye, the cells can be exposed to light to photoactivate EMA to crosslink to dead cell DNA. After crosslinking to dead cell DNA, the samples may be fixed and permeabilized. Image-IT DEAD Green Viability Stain (Cat. No. I10291) for imaging and high-content screening (HCS) analysis is a live-cell impermeant DNA binding dye that is compatible with fixation and permeabilization with good retention up to 48 hours. We also have a LIVE/DEAD Reduced Biohazard Cell Viability Kit (Cat. No. L7013) for imaging and flow analysis that contains two DNA binding dyes, SYTO 10 and Dead Red, that are sufficiently retained to be analyzed soon after 4% glutaraldehyde fixation.
Note: In general, DNA-binding dyes and calcein AM are not compatible with fixation, as these dyes are not covalently bound to components of the cell and will thus slowly diffuse out of cells after fixation, gradually staining all cells as dead.

Answer Id: E14970

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Chemical Structure

LIVE/DEAD® Reduced Biohazard Cell Viability Kit #1

COMPONENT B: DEAD Red™ nucleic acid stain
Molecular Weight: 0
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Manual / Product Insert

User Guide: alamarBlue Cell Viability Assay Reagent

Version: Jan. 2015
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  • 88951(Discontinued)
  • 88952(Discontinued)

Product Literature

Brochure: Imaging, labeling, and detection solutions

Product Literature

Brochure: EVOS Cell Imaging and Analysis Systems

Product Literature

Brochure: EVOS Imaging Systems (Chinese)

Citations & References

Automated image analysis of live/dead staining of the fungus Aureobasidium pullulans on microscope slides and leaf surfaces.

  • Authors: Nelson CD, Spear RN, Andrews JH
  • Journal: Biotechniques (2000) 29:874-80, 882
  • PubMed ID: 11056819

Manual / Product Insert

User Guide: Pierce LDH Cytotoxicity Assay Kit

Pub. No.: MAN0011851
Version: A.0

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