2022-00607-ACAN - SINGLE-CELL WESTERN SYSTEM

BACKGROUND:
Notice is hereby given by the University of Ottawa of the intent to enter a contract with Protein Simple (Bio-Techne) to procure:

• A Milo single-cell western system,
• Equipped with a two-color microarray scanner, and
• A computer including the Scout image analysis software to treat single-cell western data.

• Based on in-chip electrophoretic migration principle
• Capturing/isolating single-cells between 7 to 25 m of diameter
• Performing in-chip cell lysis
• Enabling simultaneous analysis of 1000 single cells per run
• Power requirement; 100-240 VAC input, 50/60 Hz, 2A/1A Maximum rated input current
• HV circuit rating; 300 V, 2.5 A max

• Resolution of 3 - 40μm/pixel
• Equipped with 532nm and 635nm lasers enabling the detection of Cy3, Alexa Fluor 546, Alexa Fluor 555, Cy5, Alexa Fluor 647 & Alexa Fluor 660 fluorophores.
• Detection type: Real-time confocal with 2 photomultipliers (PMT). PMT gain adjustable from 0 to 100% (linear 1 to 100%).
• Optical filter: 1 standard fluorescent filter by detection channel with real-time autofocus
  • Manual focus: offset adjustment 300 μm range, 1 μm increment
• Slide size: Compatible with all standard microscope slides: 25-26 x 75-76 mm2 / 1” x 3”; thickness: 0.9 - 1.2mm
• Scanning area: Adjustable up to 22 x 74 mm2
• Image format: TIFF (16-bit and 20-bit in dynamic extension mode)
• Electrical: ~100-240 VAC, 1.2 A, 50-60 Hz

• Hardware (Apple or PC) with 16 GB RAM, line voltage: 100–240V AC, frequency: 50Hz to 60Hz, single phase.
• 64-bit versions of Windows 7 and 10 or Mac OS-X 10.11 (El Capitan), 10.12 (Sierra), 10.13 (High Sierra).
• Equipped with the single-cell analysis Scout software v2.2.1beta (or higher).

• The single-cell western system must be based on the technology described by Hughes et al. (Nature Methods, 2014) using polyacrylamide gel chips with integrated cell-capturing micro wells, and able to achieve size-based protein separation on ~1,000 single-cells per biological sample. The system must include a high-resolution chip scanner (or equivalent technology) offering a multiplexing capacity of up to four markers per run (e.g. 532nm and 635nm lasers used to simultaneously detect 2 sets of markers with different molecular weights). The single-cell western system must enable measurement of protein targets irrespective of their location in or on a cell. It must also enable discrimination between isoforms, unspecific antigen binding (inaccurate molecular weight), and size shifts due to posttranslational and/or proteolytic processing. The cost of specialized consumables per run (excluding antibodies) must not exceed ~$400 CAD. The system must allow the use of standard commercial primary antibodies.
• The single-cell western system must run on a user-friendly image analysis software enabling automated area under the curve calculations for immunodetection signal to quantify target abundance in each of the thousand single cells analyzed per run. The software must integrate the single-cell western data to portrait cell subpopulations across the heterogeneity of a sample. The software must be available with multiple licenses to enable stand-alone desktop analysis by the users.