Flow Cytometry | Cell Sorters

In this section you will find information related with all the Flow Cytometry Sorters and respective services currently available at the Parnassus Flow Cytometry Core, these are:

  1. Self sorting on the FACSAria
  2. Self sorting on the SH800
  3. Self sorting on the WOLF
  4. Operator Assisted Cell Sorting

This LINK contains the configurations of all instruments in the Core. 

The Summary Table will provide a matrix of all the key features of each instrument. Click on the nickname link of a specific instrument to get a more detailed layout.  Tabs are also at the bottom listing the machine nicknames. It is essential for planning your experiments and seeing if an instrument can detect your specific markers. Please contact us if you have any questions or are in need of a specific filter set or setup for your experiments. 

Availability: 24 hours, 7 days a week. (Monday-Friday, 8am-7pm with "Sorter Setup.")

Setup and Operation

During normal hours of operation Monday-Friday, 8am-7pm, the sorters will be setup for you by Flow Core staff based on your needs documented on the scheduler. “Sorter Setup” is defined as: proper nozzle inserted, stable stream breakoff set, machine QC'd, Drop Delay calibrated, and sort streams aligned. After setup has been performed, if you have complete Training requirements you can run samples yourself. If you are not a SuperUser and need assistance setting up the instrument during non-normal work hours (Monday-Friday from 7pm-8am, weekends, or holidays), you can request 1 hour of Assistance so we can get the instrument setup for you. If you sort frequently during non-normal work hours, it is highly recommended to go through the Aria Advanced Training to become a SuperUser. If you sort infrequently, it is recommended to request Assistance from a Core staff for your cell sorting needs.

Training:  All Self Sorting users need to fulfill Basic Training requirements. Click HERE.

Assistance:  If you run samples infrequently, it is recommended to request Assistance to have a Flow Core staff member run samples for you. Click HERE.

Rates:  Click HERE.

Scheduling:  Click HERE.

Data Analysis:  Raw data is in a FCS file format.  For analysis options, click HERE.

 

Self Sorting on the FACSAria

The BD FACSAria2 SORP, FACSAria3u, and FACSAria Fusion are flow cytometer cell sorters equipped with 4 or more lasers, generating data of up to 20 parameters for each event passing through the interrogation point, and allows the collection of up to 4 different populations.

Although the various Aria models might look slightly different on the outside, their setup and operation is the same. These sorters can detect 2 relative physical properties as well as up to 18 different colors simultaneously. The FACSAria Fusion has an additional small particle detector for analysis and sorting of particles down to 200nm.  The Arias accommodate interchangeable nozzles ranging in sizes from 70 to 130 microns and are able to analyze and sort cells at various pressures. Analysis occurs at the cuvette flow cell, increasing sensitivity and resolution. Acceleration of the stream through the nozzle achieves droplet rates similar to that of jet-in-air sorters. Using a 70 micron nozzle, a sample can be analyzed at a rate of 25,000 events per second (90 million events per hour) and sorted with a purity of greater than 98%. Samples can be run at user-adjustable flow rates and sorts have been performed under various conditions resulting in viable cells that proliferated for several days post-sort. The sample chamber and collection tube holder can be temperature controlled and sorted samples can be collected in various tube types with up to 4 different populations sorted at one time.  In addition, all Arias in the Core have an Automated Cell Deposition Unit (ACDU) which allows for single cell sorting into plates (384 well plate max) or onto slides.  For single cell sorting, there is also an option of Index Sorting, allowing one to record the fluorescence levels of the one sorted cell, and tie that data with any downstream application. All Arias in the Core are equipped with an Aerosol Management Option (AMO) which contains aerosols that are generated during a clog.  BSL-2 samples can be sorted on these instruments.  The Arias are operated using FACSDiva Software and are available for all labs to use.

 


Self Sorting on the SH800

The Sony SH800 is a walk-up flow cytometer cell sorter equipped with 4 lasers, generating data of up to 8 parameters for each event passing through the interrogation point, and allows the collection of up to 2 different populations. 

This sorter can detect 2 relative physical properties as well as up to 6 different colors  simultaneously. The SH800 can accommodate interchangeable nozzle chips ranging in sizes from 70 to 130 microns and are able to analyze and sort cells at various pressures. Analysis occurs at the microfluidic sorting chip, increasing sensitivity and resolution.  Acceleration of the stream through the nozzle chip achieves droplet rates similar to that of jet-in-air sorters. Using a 100 micron nozzle chip, a sample can be analyzed at a rate of 10,000 events per second (36 million events per hour) and sorted with a purity of greater than 98%. Samples can be run at user-adjustable flow rates and sorts have been performed under various conditions resulting in viable cells that proliferated for several days post-sort. The sample chamber and collection tube holder can be temperature controlled and sorted samples can be collected in various tube types with up to 2 different populations sorted at one time.  In addition, the SH800 has a Sort Deposition System which allows for single cell sorting into plates (96 well plate max) or onto slides.  For single cell sorting, there is also an option of Index Sorting, allowing one to record the fluorescence levels of the one sorted cell, and tie that data with any downstream application.  The SH800 in the Core is equipped with an Aerosol Management Option (AMO) which contains aerosols that are generated during a clog.  BSL-2 samples can be sorted on this instrument.  The sorter is operated using Sony SH800 Software and is available for all labs to use.

This service is highly recommended for those who sort using 6 colors or less and those labs using fluorescent proteins. If certain laser lines are required, we’ll help you make a recommendation on which instrument would be best for your situation.

 


Self Sorting on the WOLF

wolf_and_n1DEMO ENDING AUGUST 2020.

The WOLF Cell Sorter uses patented, microfluidic-based sorting and with robust laser detection and sensitive PMT detection compatible with mammalian cells, microbes, plant calls and more. A gentle and precise on-chip piezoelectric actuator directs cells into collection channels, and allows analysis and sorting in a disposable format. This eliminates sample-to sample contamination and biohazard exposure or cleanup.

At <2 psi, the WOLF is gentler than any conventional cell sorter, enabling healthier cells post sort, especially for engineered lines, primary cells and stems cells. Intuitive software, fixed optics, no fluidics cart and less than one-minute clean-up time. Disposable, aerosol-free microfluidic cartridge allows for sterile sorting that protects the sample from the environment and the scientist from the sample. At under 2 cubic feet, NanoCellect’s benchmark for access and performance allows every lab the flexibility to do analysis and sorting into tubes or 96- and 384-well plates.

WOLF Demo Days:  First Tuesday of every month from 10am - 5pm starting May 7th, 2019  - come demo the WOLF Cell Sorter and N1 Single Cell Dispenser with NanoCellect's Field Application Scientist. The WOLF offers gentle sorting with disposable microfluidics to preserve viability and guarantee sterility. Please RSVP here if you would like to schedule an appointment.  

Visit NanoCellect website for more information.

 


Operator Assisted Cell Sorting 

On other Cell Sorters

Service available for any Cell Sorter by requesting Assistance Service.

On the MoFlo / BD Influx

Availability: Monday-Friday, 10am-5pm. Core Staff Operator must be present.   

Overview:  The BD Influx flow cytometer cell sorter is equipped with 2 lasers, generating data of up to 8  parameters for each event passing through the interrogation point, and allows the collection of up to 4 different populations.