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Multicolor Labeling of Dead Cells for Flow Cytometric Analysis


Detection of Jurkat cell viability using Cell Explorer™ Fixable Dead Cell Staining Kit. Jurkat cells were treated and stained with Stain IT™ Green and then fixed with 3.7% formaldehyde and analyzed by flow cytometry. Live (blue solid peak), staurosporine treated (green line) and heat-treated (red solid peak)cells were detected with FL1 channel (Ex/Em = 488/520 nm). Nearly identical results were obtained before fixation.
The effective labeling of cells provides a powerful method for studying cellular events in a spatial and temporal context. AAT Bioquest Cell Explorer™ Fixable Dead Cell Staining kits are a set of tools used to label cells for fluorescence microscopic and flow cytometric investigations of cellular functions.

Cell Explorer™ Fixable Dead Cell Staining Kits employ cell components-reactive fluorescent stains to evaluate mammalian cell viability using flow cytometry and fluorescence microscope. In cells with compromised membranes, the stains react with cell components both in the cell interior and on the cell surface, yielding intense fluorescent staining. The proprietary stains used in the kits become more fluorescent upon binding to cellular components. These cell stains are not live cell-permeable. In viable cells, the stain's reactivity is restricted to the cell-surface components, resulting in less intense fluorescence. The difference in intensity between the live and dead cell populations is quite large, and the fluorescence intensity discrimination is completely preserved following formaldehyde fixation. Moreover, these stains use only one channel of a flow cytometer, making them compatible with multiparameter staining experiments for multiplexing applications.

The fluorescence signals of the stains used in the kits are pH-independent and quite photostable. The stains have much better water solubility, making the kits easier to use. Cell Explorer™ Fixable Dead Cell Staining Kits provide all the essential components with an optimized fixable dead cell staining protocol that requires minimal hands-on time.

Live HeLa Cells
Fixed HeLa Cells

HeLa cells stained with Cell Explorer™ Fixable Dead Cell Staining Kit in a 96-well Costar black wall/clear bottom plate.


Key Features of Fixable Dead Cell Staining Kits

  • A full spectrum of colors available for multiplexing
  • Compatible with either flow cytometry or fluorescence imaging
  • Minimal hands-on time required
  • Robust performance for tracking cells

 

Table 1. Live or Dead™ Fixable Dead Cell Staining kits for live/dead cell analysis during flow cytometry

Kit
Laser (nm)
Ex/Em (nm)
Multiplex
Fixable
Unit Size
Cat No.
Live or Dead™ Fixable Dead Cell Staining Kit *Blue Fluorescence*UV (350 nm)353/442YesYes200 Tests22600
Live or Dead™ Fixable Dead Cell Staining Kit *Blue Fluorescence with 405 nm Excitation*Violet (405 nm)410/450YesYes200 Tests22500
Live or Dead™ Fixable Dead Cell Staining Kit *Green Fluorescence with 405 nm Excitation*Violet (405 nm)408/512YesYes200 Tests22501
Live or Dead™ Fixable Dead Cell Staining Kit *Green Fluorescence*Blue (488 nm)498/521YesYes200 Tests22601
Live or Dead™ Fixable Dead Cell Staining Kit *Orange Fluorescence with 405 nm Excitation*Violet (405 nm)398/550YesYes200 Tests22502
Live or Dead™ Fixable Dead Cell Staining Kit *Orange Fluorescence*Green/Yellow (532/561 nm)547/573YesYes200 Tests22602
Live or Dead™ Fixable Dead Cell Staining Kit *Red Fluorescence*Yellow (561 nm)583/603YesYes200 Tests22603
Live or Dead™ Fixable Dead Cell Staining Kit *Red Fluorescence Optimized for Flow Cytometry*Blue/Green (488/532 nm)523/617YesYes200 Tests22599
Live or Dead™ Fixable Dead Cell Staining Kit *Red Fluorescence Optimized for Flow Cytometry*Red (635 nm)649/660YesYes200 Tests22604
Live or Dead™ Fixable Dead Cell Staining Kit *Red Fluorescence Optimized for Flow Cytometry*Red (635 nm)749/775YesYes200 Tests22605