Journal: Scientific Reports

Article Title: Rapid identification and drug susceptibility screening of ESAT-6 secreting Mycobacteria by a NanoELIwell assay

doi: 10.1038/srep00635

Figure Lengend Snippet: (1) The nanoELIwell master was fabricated using silicon wafers and standard rapid prototyping techniques. (2) PDMS soft photolithography was employed to generate the elastomeric nanoELIwells for bacterial cultures. (3) The PDMS was then peeled off from the master and sized if needed before each experiment. The nanoELIwells were then placed on top of an acrylic plate with holes, which allow for the delivery of oxygen/CO 2 during culture. (4) Bacteria were deposited onto oxygen plasma-treated or fibronectin-soaked nanoELIwells and covered by the antibody-coated glass slide. (5) The device was then held together with a second acrylic plate (without holes) and cultured for 24–48 hours at 37°C (as shown in Figure A) before being dismantled for analysis. (6) The glass slide was carefully removed without disturbing the bacteria in the nanoELIwells. (7) The ELISA assay was conducted with a fluorescently-labeled secondary antibody and imaged using a microarray scanner (Figure B). (8) The nanoELIwells PDMS stamp were completely dried under a vacuum, coated with a thin layer of gold, and imaged using SEM techniques (Figure C).

Article Snippet: SuperEpoxy2, Protein Printing Buffer, Washing Buffer, Rinsing Buffer, Reaction Buffer, BlockIT, and the Microarray High-Speed Centrifuge used for microarray analysis were purchased from ArrayIt (Sunnyvale, CA, USA).

Techniques: Bacteria, Clinical Proteomics, Cell Culture, Enzyme-linked Immunosorbent Assay, Labeling, Microarray

Journal: Scientific Reports

Article Title: Rapid identification and drug susceptibility screening of ESAT-6 secreting Mycobacteria by a NanoELIwell assay

doi: 10.1038/srep00635

Figure Lengend Snippet: EAST-6 secreting M. kansasii presented intensive signals on NanoELIwells coated with ESAT-6 antibody (A), but no signal from non-ESAT-6 secreting M. simiae (E) from a 48 hour nanoELIwell culture, insets are zoomed-in images. The inset in (E) displays 5 μm resolution pixels of the microarray scanner limit, indicating only background noise and no detectable signal. SEM images (B–D) of nanoELIwells contained M. kansasii in 100x100 μm ELIwells and SEM images (F–H) contains M. simiae in 50x50 μm ELIwells.

Article Snippet: SuperEpoxy2, Protein Printing Buffer, Washing Buffer, Rinsing Buffer, Reaction Buffer, BlockIT, and the Microarray High-Speed Centrifuge used for microarray analysis were purchased from ArrayIt (Sunnyvale, CA, USA).

Techniques: Microarray