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Cell Sorting

Please use the Scheduling Smartsheet Request Form to schedule a sorting reservation

The Dana Farber Flow Core offers cell sorting performed by experienced cytometry technicians. Please follow the instructions below to optimize your sorts. See the Rates for Sorting and Instruments pages for more information.

Download the Before Your Sort Guide

Reference the "Before You Sort" PDF for all things sorting at the Flow Cytometry Core at Dana-Farber Cancer Institute.

Sample Preparation

Samples MUST BE FILTERED no more than 30 minutes before the start of your reservation. Please use 35 mm filter cap FACS tubes if available.

Please prepare your cells at the appropriate concentration:

  • 70 µm nozzle - 10-15 million/mL or less
  • 100 µm nozzle - 10 million/mL or less
  • Plate sorting or adherent cells - 5 million/mL or less

Cells can be prepared in any media or PBS. Please bring collection tubes with media or buffer in the bottom.

Controls are required for the technicians to properly set up your sort. Please bring a negative/unstained control, and single color controls for each fluorochrome in your experiment.

Nozzles

The nozzle generates droplets containing cells that the instrument then sorts. We offer two different nozzle sizes:

70 µm Nozzle

  • Default nozzle​
  • 70 PSI
  • Useful for most cells types
  • 1 mL of recovery volume ≈ 1 million cells

 

100 µm Nozzle

  • Requires 10 minutes at the beginning and end of your reservation to switch nozzles
  • 20 PSI
  • Best for large, fragile, or adherent cells
  • 100 µm Nozzle is best for sorting into plates
  • 3 mL of recovery volume ≈ 1 million cells

Please select the nozzle you are using on the iLabs reservation sheet so we can set up before you arrive if time allows.

Sort Collection

Sorted cells can be collected into:​

  • 0.5 - 2 mL Eppendorf Tubes
  • 5 mL FACS Tubes
  • 15 mL Conical Tubes
  • 6, 12, 48, 96, or 384 Well Plates (Culture or PCR)

Please make sure to have media in your collection tubes, and choose collection tubes based on the number of cells you expect to get back per sample and the nozzle you are using:

70 µm Nozzle

  • < 500,000 cells - Eppendorf tube with 500 uL of media
  • 500,000 - 3,000,000 cells - 5 mL FACS tube with 1 mL media
  • > 3,000,000 cells - 15 mL tube with 3 mL media

100 µm Nozzle

  • < 250,000 cells - Eppendorf tube with 500 uL of media
  • 250,000 - 1,000,000 cells - 5 mL FACS tube with 1 mL media
  • > 1,000,000 cells - 15 mL tube with 3 mL media

Note: ​The number of cells that are collected into the tube are usually 60 - 80% of the number that the instrument reports as sorted. Please take this into consideration when planning your experiments.

Lentivirus and Human Primary Cells

Lentiviruses can deliver a significant amount of viral RNA into the DNA of the host cell and have the unique ability among retroviruses of being able to infect non-dividing cells, so they are one of the most efficient methods of a gene delivery vector. HIV, SIV, and FIV are all examples of lentiviruses.

Sorting Lentivirus and Human Primary Cells

Requirements:

  • Bring a red Biohazard disposal bag to the lab at the beginning of you sort.
  • Dispose of the  biohazard bag at the end of your sort.
  • Samples must also be contained in closed cap tubes (No Filter Caps). Lentivirus and HPC samples will only be sorted if you meet these requirements. 

Read the full Lentivirus Sorting protocol here.

Read the full Human Primary Sorting protocol here.

Sorter Training

Sorter training for the Sony MA900 is available for all users. Please contact the core for more information.