The % cell viability is not provided when using this app.
The PI app uses brightfield images to capture live cell counts and red fluorescence images for
counting dead cells.
Propidium iodide (PI) is a red-fluorescent nuclear and chromosome stain that only enters dead cells.
Upon entry, the PI stain binds to DNA by intercalating between the bases with little or no sequence
preference. In aqueous solution, the dye has excitation/emission maxima of 493/636 nm. When
bound, the excitation/emission maxima shift to 535/617 nm respectively.
Calculation:
% Viability =(Brightfield count / (Brightfield count + PI count)) *100.
The AO/PI App uses green fluorescent images to capture live nucleated cell counts and red
fluorescent images for counting dead cells. The application is suited for counting cell culture samples
or primary samples.
Acridine orange (AO) and propidium iodine (PI) are both cell permeable nuclear dyes that in
combination are used to count live and dead cells within a cell solution. The AO enters both live and
dead cells and results in green fluorescing cells. PI enters only dead, nucleated cells with
compromised membranes to generate red fluorescence. Cells stained with both AO and PI fluoresce
red due to fluorescence resonance energy transfer (FRET).
% Viability =(AO count / (AO count + PI count)) *100
The GFP App uses green fluorescent images to count GFP transfected cells and brightfield images to
count non-transfected cells
As GFP intensity can be highly variable it is recommended that the fluorescent threshold is set for
each sample after the count using the intensity slider in the results screen.
The green fluorescent protein (GFP) isolated from the Aequorea victoria jellyfish is a useful
reporter molecule for monitoring gene expression. GFP emits green light (lambda max = 509 nm)
when excited with UV or blue light resulting in cells that fluoresce green.
The GFP app is used to determine transfection efficiency of transient transfections.
% Efficiency =(GFP+ count / (GFP+ count+ Brightfield count)) *100
The Yeast App uses green fluorescent images to capture live yeast cell counts and red fluorescent
images for counting dead yeast cells.
Yeast cell live/dead staining Is performed using fluorescein diacetate (FDA) and propidium iodide (PI).
FDA freely moves in and out of the plasma membrane of diverse organisms from bacteria to
mammalian cells. The non-fluorescent FDA taken up by cells is converted the into the green
fluorescent metabolite fluorescein. The green fluorescence is used as an indicator of live cells. PI
enters only nucleated cells with compromised membranes to generate red fluorescence indicating
dead cells.
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