Equipment_Specifications_.pdf

Equipment Specifications

This piece of equipment will be utilized for projects within the Chemical Safety for

Sustainability (CSS) workplan for NCCT and potentially cross divisional work within EPA for collaborators that need cells sorted based on fluorescent activity level. As such, it will need to have the following specifications to meet the demands of the laboratory and users.

-Has a droplet frequency of 37-43 kHz.. This is needed for proper function of the equipment.

-Uses true jet-in-air for high-performance sorting.

-Has no hardware limitations for sort rate, which is limited only by droplet frequency and application. This is needed so that the machine can be flexible for experimental design needs.

-Can sort cells at up to 99% purity. This is needed so that there will not need to be multiple sort steps and so that investigators can trust the sorted cells are viable and ready for use in experiments.

-Has three different sort collection modes: single, purity, and enrich. This is needed for flexibility in user experimental design.

-Automatically calculates drop delay. This is needed so that there does not need to be any user intervention upon startup and calibration of the unit.

-Automatically aligns stream to laser upon startup without user intervention. This is needed since this machine will be a multiple user piece of equipment, the automation will allow multiple users to easily utilize the machine.

-Automatically monitors and maintains droplet break-off during sort. This is needed so that the final sorted product is of the highest quality.

-Automatically align nozzle tip and stream to optics during nozzle tip swaps. This is needed so that there is not a need for user intervention.

-Uses five picomotor controls, which are completely driven by software, to make micrometer adjustments to the nozzle stage. This is needed so that the unit will automatically adjust for the cleanest cell sort.

-Utilizes three cameras to monitor fluidics, stream alignment, and droplets, which helps the software automate the system and make fine adjustments in real time. This is needed for the equipment to have the cleanest cell sort possible without user intervention.

-Has 488 nm 100 mW and 488 with either 561 nm 100 mW or 640 nm 100 mW Class 1 lasers.

This is needed because multiple fluorescent protein colors are utilized across CSS experiments.

-Utilizes a fixed 100 μm nozzle tip. This is needed for the correct droplet formation during sorting

-Has an onboard sheath fluid dilution system for longer run times between sheath fluid exchanges. This is needed to ensure the same quality from run to run and user to user since there is not a need to completely flush out the system and refill with solution every run.

-Includes a 4 L container of Sheath Fluid, which provides 50 hours of run time, and a 4 L container of deionized water, which provides approximately 9 hours of run time. This is needed to allow for continual runs without the hassle of changing fluids within runs or everyday.

-Allows uninterrupted fluidic exchanges during the sort run without the need to repressurize the system. This is needed for shorter run times if the fluids do need replaced during a run.

-Has easy-to-exchange filter blocks, which can be customized for unique fluorophore panels.

This is needed to ensure that this machine is able to be utilized for flexible experimental designs in the future vs. a fixed system.

-Automatically washes the sample line with each sample exchange using the unique loading stage. This is needed to ensure there is no cross contamination between cell sorting runs.

-Uses true vortexing agitation in the sample station to ensure the samples stay in suspension.

This is needed to ensure that the cell sorting run is efficient and that all the cells from the sample are sorted.

-Includes an onboard Peltier solid state temperature control system; temperature can be maintained from 4 to 37°C to heat or cool the sample station and sample collection area. This is needed for sample stability during cell sorting.

-Has a strategically placed bubble detector, which allows detection of end of sample without introducing bubbles to the fluidic stream. This is needed to ensure quality of the run. If a bubble gets into the system and is not detected, the droplets cannot be verified for the correct fluorescent protein and the purity of the run will be affected.

-Maintains user safety from lasers and sort deflection plates with safety interlocks at the nozzle door and sort chamber sliding door. This is needed to ensure safety of laboratory personnel.

-Sorts and analyzes data, recalls data files and creates protocols for experiments. Also prints user, daily quality control (QC), or QC trending reports, which can all be saved as distinct .csv files. This is needed so that the user can verify and have a quality record of the cell sorting run.

-Has dimensions of no more than 70 x 65 x 65 cm (2.3 x 2.1 x 2.1 ft) (W x D x H); weight is 90 kg (198 lb).. Due to size constraints in the laboratory, the fluorescent activated cell sorter must be small enough to reside on our current laboratory bench depth, which is 29.5 inches.

-Uses three simple connections: Ethernet and USB to computer, with standard 110 V and 220 V electrical outlet. This is needed for ease of data transfer and to accommodate current electrical setup in the laboratory