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Lab Pre-Flight

eshimabAbout 2 min

Yesterday: Prep Sheath

Create the sheath fluid the day before

  • If Using BioSure or MilliQ for Sheath fluid, this is handy
  • If dissolving NaCl into MilliQ, it is required.
  • See section on Sheath Prep.

Run Day

House Air

Always check the house air valve first!

  • The InFlux air pressure is provided by an air compressor in C-MORE's attic.
  • If the building power flickers, the air compressor goes offline.
  • Only UH Facilities have the key to the compressor room.
  • We are checking the compressor first because, if it is off, the instrument can't be run until UH Facilities restarts the compressor.

asset "House Air Valve"

The House Air valve is under the lowest shelf on the bottom right corner of the bench opposite the instrument.

  1. Open the valve on the wall opposite the InFlux Chassis to access the building's air compressor (i.e. House Air).
    1. You should hear a hiss as the tubing pressurizes.
    2. If no air flows, then the compressor may be off. This often happens if the building loses power. If the compressor is off:
      1. Call UH Facilities at 1-808-956-7134 and tell them that they need to restart the building compressor at C-MORE.
      2. Tell them that this is impeding work and needs to be done asap.

Fill Sheath Tank

asset "Sheath Tank Prep Picture"

Picture of the graduated cylinder and/or salt and/or MilliQ.

  1. Use Graduated Cylinder to Mix Sheath Fluid
    1. For Non-Sorting, pure MilliQ Water is fine.
    2. For Sorting, we need a saline solution:
      1. For BioSure: Mixture is 1/8 BioSure Concentrate and 7/8 MilliQ Water in a Graduated Cylinder
      2. For NaCl: Dissolve 32g NaCl in 4 liters MillQ Water. Let Sheath Fluid sit overnight to fully dissolve NaCl
    3. Fill a 60ml Syringe with Sheath Fluid for Purging the instrument and filling sort tubes.
      1. Label Syringe.
      2. Attach a 0.2um Acrodisc Syringe Filter to the syringe

Syringes

asset "Filled Syringes"

Three syringes filled with sheath fluid, MilliQ Water, and 70% ethanol.

  1. Sheath Syringe
    1. Fill a 60ml Syringe with Sheath solution using the LuerLok Adapter.
    • Make sure the Sheath Tank is not sealed.
    1. Attach a 0.2um Acrodisc filter.
  2. MilliQ Syringe
    1. Fill a 60ml Syringe with MilliQ Water.
    2. Attach a 0.2um Acrodisc filter.
  3. 70% Ethanol (sometimes useful)
    1. Fill a 60ml Syringe with 70% Ethanol.
    2. Cap the Syringe.
    3. Ethanol will eat away at the Acrodisc filter. Install the filter only when necessary.

Alignment/Calibration Samples

asset "Alignment Sample Tubes"

Filtered MilliQ, HB-` "Beer", 1.0um bead dilution, and (if sorting) 3.1um bead dilution.

  1. Filtered MilliQ
    • From the MilliQ Syringe with 0.2um Filter
  2. Coarse Alignment "Beer"
    • Fill a sample tube with about 800ul of "HB-1" Beer.
    • This is a highly toxic lab chemical. Do not consume.
  3. Laser Alignment 1.0um Calibration beads
    • Create a dilution of 1.0um Ultra Rainbow Fluorescent Beads (Spherotech) by dropping 2-3 drops of Bead Concentrate into ~2ml 0.2um Filtered MilliQ Water
      • Note: The 1.0um Bead concentration is 1E7
  4. Drop Delay 3.1um Calibration Beads
    • Create a dilution of 1.0um Ultra Rainbow Fluorescent Beads (Spherotech) by dropping 5-6 drops of Bead Concentrate into ~2ml 0.2um Filtered MilliQ Water.
      • Note: The 3.1um Bead concentration is 1E6 (lower than the 1.0 um)
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