Get a Yes/No Fast without the Flow Core:

Gain your Independence with the CellSimple™ Cell Analyzer

CellSimple

The CellSimple™ Cell Analyzer is a benchtop instrument that combines fluorescence, cell counting, and volume measurements with single-use cassettes to perform powerful "on-demand" cell and bead-based assays.

The system is:

  • Fast – Accurate results in less than a minute right at your bench!
  • Simple – No fluidics or flow cell to maintain. No extensive training to operate.
  • Powerful – Bead and cell-based experiments at the push of a button.

The CellSimple™ Cell Analyzer is currently only available in the US.

Pathway Activation Assays Using Phospho-specific Antibodies

Download the Application Note for more information.

Jurkat cells were treated according to the table below, and prepared using the CST protocol for intracellular flow-cytometry. The cells were then double-stained with Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (197G2) Rabbit mAb (PE Conjugate) #14095, and Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb (Alexa Fluor® 488 Conjugate) #4071, using standard CST protocols for immunofluorescent analysis. The samples were assayed using the Open Flow Application on the CellSimple™ Cell Analyzer. Fluorescence emission from phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (197G2) Rabbit mAb (PE Conjugate) was measured using PMT1 (561 nm/ LP) and Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb (Alexa Fluor® 488 Conjugate) was measured using PMT2 (525/45 nm) and the results are summarized in the table.

Pathway Activation Assays Using Phospho-specific Antibodies
Panel Treatment Pathway Status Result
A LY294002, wortmannin, U0126 PI-3K/Akt = Off
MAPK = Off
Neither Akt nor p44/42 MAPK (Erk1/2) is phosphorylated, so the fluorescent signal in both channels is low.
B U0126 PI-3K/Akt = On
MAPK = Off
Akt is phosphorylated, so the fluorescent signal in PMT2 (x-axis) is high. p44/42 MAPK (Erk1/2) is unphosphorylated, so the fluorescent signal in PMT1 (y-axis) is low.
C (LY294002, wortmannin) > PMA PI-3K/Akt = Off
MAPK = On
Akt is unphosphorylated, so the fluorescent signal in PMT2 (x-axis) is low. At the same time, p44/42 MAPK (Erk1/2) is phosphorylated, so the fluorescent signal in PMT1 (y-axis) is high.
D PMA PI-3K/Akt = On
MAPK = On
Both Akt and p44/42 MAPK (Erk1/2) are phosphorylated, so the fluorescent signal in both channels is high.

These data indicate that the CellSimple™ Cell Analyzer can be paired with antibodies against phosphorylated proteins to measure independent activity changes in multiple signaling pathways at the cellular level.

Assess Signaling Pathway Activity Using Bead-conjugated Antibodies

Download the Application Note for more information.

Assess Signaling Pathway Activity Using Bead-conjugated Antibodies

Instrument screen shot showing four separate populations of beads (representing separate targets) in the presence (above the red line) or absence (below the red line) of Calyculin A (+Cal A) treatment in the panel on the left. Data was exported from the instrument and plotted in the panel on the right.

This assay is founded upon the sandwich immunoassay principle. Cell lysates are incubated with a capture antibody bead cocktail, which contains target-specific antibodies individually conjugated to beads of different sizes. After incubation with the lysate, the capture beads are washed and incubated with a biotinylated detection antibody cocktail. Finally, phycoerythrin-conjugated-Strepdavidin is used to visualize the bound biotinylated-detection antibody.

The Open Flow Application on the CellSimple™ Cell Analyzer was used to resolve four populations of beads based on size in both the untreated and Calyculin A treated HeLa cell lysates. The plot on the left shows an overlay of the data from the untreated cell lysates (below the red dashed line) and the treated cell lysates (above the red dashed line). These data are plotted in the graph on the right and indicate that phosphorylation of each of the target proteins is stimulated in response to Calyculin A treatment.

These data demonstrate that the CellSimple™ Cell Analyzer can support multiplexed, bead-based analysis of signaling pathways.

Cell Health Assays Using Cellular Dyes

Download the Application Note for more information.

Cell Health Assays Using Cellular Dyes

Jurkat cells were either untreated (left panel) or treated with staurosporine (right panel). The cell count, MFI and and percent of total population for each panel can be visualized at the corner of each gate.

Jurkat cells were either left untreated (left) or treated with staurosporine (10 μM, 18 hr; right) and then stained with Propidium Iodide (0.5 μM) and Calcein-AM (0.05 μM). Treatment with staurosporine induces cell death, shifting the cellular population from the lower right hand quadrant (live cells) to the upper left hand quadrant (dead or dying cells) of the plot. The Cell Health Application on the CellSimple™ Cell Analyzer automatically runs data analysis to quantify the shift, revealing a drop in living cells from 88.2% in the untreated culture (left, green circle) to 1.14% in the staurosporine-treated culture (right, green circle) and an increase in dead cells from 6.29% in the untreated culture (left, red circle) to 84.2% in the staurosporine-treated culture (right, red circle).

These data demonstrate that the Cell Health application on the CellSimple™ Cell Analyzer used in conjunction with Calcein-AM and PI in one assay allows a researcher to measure the health of a cell sample quickly and accurately.