ACR

Autonomous, capillary-flow, microscope-slide-based Cell Retainer (ACR)

The Cell Retainer (CR) approach serves as a conceptual basis for a variety of individual cell based assays.

The CR is a densely packed 2-D arrangement of hexagonal Pico-liter Wells (PW) ~ 8µm depth, 20µm pitched, made of glass, in a honeycomb-like pattern. Each PW, designed to contain a single un-tethered cell, functions as a high-quality concave micro-lens.

The PW's internal surface is smooth, preventing light scattering; its bottom is nearly flat, minimally diverting the wave front of transmitted light. The walls edges are extremely sharp (less than 0.1 µm wide), causing the precipitating cells to settle inside the wells rather than in between. The 6 sides of a PW are semi-lunar, allowing for the flow of solutions, while confining the freely moving cell within it.

The CR concept was embodied in several operative types of CR-bearing devices, including an Autonomous, capillary-flow, microscope-slide-based CR (ACR). This product was comercialized by Molecular-Cytomics Inc. (http://www.molecular-cytomics.com) under the name Optical LiveCell Array (LCA).

The ACR enables repetitive, high-content signal and image analysis of the same non-adherent, non-tethered individual cells or groups of cells, being subjected to various bio-manipulations and staining procedures, while maintaining cell viability and identity for hours. Embedded under a 0.7mm deep, 2mm cylindrical aperture drilled in an engraved plastic plate, the CR offers about 7800 free PWs. The slide dimensions are similar to those of a standard microscope slide (76mm x 26mm), thus it is suitable for both upright and inverted microscopy, utilizing standard microscope stages.
The unique capabilities of the ACR are best exemplified by performing experiments correlating static and/or kinetic functional measurements with post-fixation parameters in the same non-adherent un-tethered cells. These kinds of investigations would be difficult, if not impossible, to conduct by other state of the art techniques, due to their inability to preserve cell identity without tethering.

 

Part 1: Macro view of liquid flow within the LCA chamber

[KGVID width="640" height="360"]http://www.jsc.ph.biu.ac.il/wp-content/uploads/Filling-Cell-Chamber-and-Capillary-Flow-in-LCA.mp4[/KGVID]

 

Part 2: Macro view of liquid exchange within the LCA chamber

[KGVID width="640" height="360"]http://www.jsc.ph.biu.ac.il/wp-content/uploads/FLOW-AND-SOLUTION-EXCHANGE-WITHIN-THE-LCA-VIA-A-BINOCULAR-MICROSCOPE.mp4[/KGVID]

 

Part 3: Cell loading and solution exchange within the LCA chamber

[KGVID width="640" height="360"]http://www.jsc.ph.biu.ac.il/wp-content/uploads/VISUALIZATION-X20-OF-CELL-LOADING-AND-TREATMENT-WITHIN-THE-LCA-VIA-INVERTED-EPI-FLUORESCENT-MICROSCOPE.mp4[/KGVID]