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Technology

Applications

Data for ELISA assays

A highly sensitive and robust ELISA platform

Our customer compared the sensitivity and robustness of a model ELISA test from a conventional 384-well microplate and a corresponding DropArray™ plate. In the test, various levels of a secreted human cytokine were measured in presence of human serum. Figure 1 displays the results of the model ELISA test. The results demonstrates that the detection obtained by Curiox's DropArray™ D384 plate is comparable in sensitivity and robustness to a conventional microplate. This shows that the DropArray™ is a highly sensitive and robust platform for performing an ELISA test.

Fig. 1A Logistic (4-param) Regression fit of Human Serum ELISA on 384 DropArray™ plate.

Fig. 1A 4-parameter logistic regression of a dose escalation ELISA assay detecting a secreted human cytokine.

Fig. 1B Plot of Percent Difference from 0%-HS by Buffer in Human Serum ELISA

Fig. 1B Comparably low CV throughout different human serum content observed using DropArray™ platform.

Data for Cell-based Assays

Robust high-content screening performance

The DropArray™ cell platform is an ideal and robust platform for statistically stringent high content assays. Figure 4 below shows an example of a Docetaxel dose escalation mitotic arrest assay performed in HeLa cells utilizing the DropArray™ D384 plate system. As observed, independent plate replicates show highly similar and reproducible EC50 values, as well as high Z'-factor values.

Fig. 4A Logistic (4-param) Regression fit of Mitotic Arrest induced by Docetaxel

Fig. 4A A 4-parameter logistic regression of a 10-point dose escalation experiment of Docetaxel in HeLa cells.

Fig. 4B Z-Prime by plate replicates

Fig. 4B Z-factor calculation for different replicates for Mitotic Arrest experiment

Fig. 4C Composite Mitotic Index Docetaxel HeLa

Fig. 4C Composite 10x microscopy image from GE IN Cell Analyzer 2000 for HeLa cells arrested in mitosis following treatment with 50 ng/ml Docetaxel for 18 hours and stained for Ph-Histone-H3,s10 (Alexa 488-green) and DNA (Propidium Iodide-red)"

Gentle cell washing: ideal for difficult to handle suspension and
semi-adherent cells

Figure 2 shows minimum detachment of semi-adherent HEK-293T cells, after performing a conventional DNA stain protocol (DAPI or Propidium Iodide) protocol on a DropArray™ D384 plate by the DropArray™ Accelerator HT200.

Fig. 2A 10x immuno-fluorescent image ofHEK-293T cells stained with DAPI DNA dye.

Fig. 2B 10x immuno-fluoresecent image of HEK-293T cells stained with Propidum Iodide.

Easy on-plate transfection for complex molecular biology screening:

Figure 3 shows a composite immuno-fluorescent microscopy image of a typical bait/capture protocol performed on the DropArray™ cell 384 "well" plate. In this example, a bait protein is transiently transfected into HEK-293T cells, and following 48 hours expression a capture protein is co-incubated with the cells. The receptor/ligand binding interaction is imaged utilizing an antibody against the capture protein.

Fig 3. 5x immuno-fluorescent microscope image of a bait/capture protein assay in HEK-293T cells performed on DropArray™ D384 plate, and washed with DropArray™ Accelerator HT200 system and imaged on a GE IN Cell Analyzer 2000 High Content Imager.

(A) DNA stain (Hoechst)

(B) FITC (Alexa-488)

Fig. 3A DNA (Hoechst)

Fig. 3B Bait/Capture (Alexa-488)

Fig. 3C Composite