Sony SH800 User Manual

January 11,2018
1
Sony SH800 Cell Sorter
SSFF
NEW
ACCOUNTS MUST BE ADDED on BEFORE THE CHIP IS SCANNED
Currently the 70, 100 and 130 µm chips are in use
Notes:
• Make sure to draw all your gates and name everything before you record your data. Once you have recorded data
you cannot draw new gates. You may copy and paste regions to new tubes.
• For sort targeting with real cells
o Set the number of events to sort higher than needed
o Test placement of sorted cell by initiating SORT
o Pause and look at deposition
o It will only do one sort per tube, so if you need to do multiple tests you do not want to reach desired # of events or
it will insist on a new tube being created
1. Start up
NOTE: The upper lid on the cytometer has pneumatic lifters, so attempting to open this with the air line
vented will create an error. The air to the cytometer should be restored before attempting to load a chip, or to
look at filters. The air line
Check the LCD display for pressure reading before opening the chip loader flap door.
1. Decontaminate and clean all surfaces of the collection chamber. And clean sort collection device if desired.
2. No Cavicide should be used on the plates. A 70% EtOH wipe daily should be sufficient. The lower
portion of the sort chamber can be wiped with a small amount of Cavicide.
i. For the 70µm chip
o Microbubbles are always a greater problem for the 70um than other nozzles and may be contribute
to the instability. In the AM, the air line vent should be closed and the vent valve on the tank turned to
reseat the seal.
o This nozzle is VERY sensitive and side streams will not form if the plates are not clean and dry
o IF sheath pressure issue occurs, check the lid of the sheath tank.

Sony SH800 Specifications

General

Type	Cell Sorter
Software	Sony SH800 Software
Category	Laboratory Equipment
Sample Input	Tubes or plates
Laser Options	Up to 4 lasers

Summary

Start Up Procedures

Decontaminate and Clean Collection Chamber

Clean collection chamber and sort collection device. Use 70% EtOH on plates.

Sheath Tank and Filter for 130µm Chip

Ensure filled sheath tank, de-bubble filter for stable stream.

Turn On Pressure

Activate pressure before proceeding with subsequent steps.

Power Up SH800 Instrument

Turn on the SH800 instrument and observe fluid levels on display.

Prepare Sheath Tank

Disconnect airline, release pressure, and fill sheath tank carefully.

Empty and Bleach Waste Tank

Empty waste tank at sink, add bleach to carboy before replacing.

Standby Mode Requirement

Use standby mode for emptying waste or filling sheath to avoid recalibration.

Fill Di Water Tank and De-bubble Filter

Fill Di water tank, then de-bubble the filter using software.

Log In to Windows

Log into the Windows operating system using the FACS User account.

Start SH800 Software

Launch the SH800 software after the instrument enters standby mode.

Log In to SH800 Software

Log into the SH800 software with user credentials; use 'facs user' if needed.

Activate Aerosol Management System

Enable aerosol management system for BSL-2 samples.

Complete Biohazard Form

Fill out the biohazard form detailing sample hazard and cleanup.

Automated Alignment and Sort Setup

Load and Exchange Chip

Scan and load a new chip; reuse old chip if <24h old.

Initial Calibration Steps

Select blue laser for calibration, then run autocalibration with beads.

Sorting Setup and Configuration

Sample Filtering Requirement

All samples, including compensation controls, must be filtered.

Experiment and Parameter Settings

Select experiment, name, and configure fluorescent parameters.

Laser Activation

Turn on 488nm laser; use violet laser sparingly to preserve chip life.

Compensation Matrix Setup

Use compensation wizard or manual method with single-color controls.

Acquisition and Sorting Workflow

Detector and Threshold Settings

Adjust FSC/BSC gain and set detector thresholds for accurate cell acquisition.

Acquire Sample Events

Acquire sufficient events for setting gates and gain adjustments.

Create and Adjust Gates

Create plots, double-click gates, and adjust settings for desired populations.

Sorting into Collection Tubes

Select sort mode, set sort gates, place tubes, and start sorting.

Plate Sorting Procedure

Prepare for 96-Well Plate Sorting

Install splash guard, select 96-well plate method, and pause for gate setup.

Plate Adjustment and Verification

Adjust plate drop positions, verify cell distribution, and re-adjust if needed.

Set Up and Run Plate Sort

Clear wells, set sort criteria, click 'Sort and Record Start'.

Analyze Index Sort Data

Click Worksheet Tools and 'Analyze Index Data' to review sort results.

Instrument Shutdown and Cleanup

Standby Mode Activation

Put the machine in standby mode via settings before maintenance tasks.

Data Export

Export FCS files and save PDF of experiment layout and gates.

Routine Instrument Cleanup

Perform bleach clean and DI water rinse cycles after use.

Log Out and Calibration Retention

Log out, checking 'Keep sort calibration' to avoid recalibration.

Advanced Cleanup and Safety

BSL-2 Sample Cleanup

Use aerosol management system, keep sort door closed, and manage suction.

Instrument Manual Shutdown

Perform hardware/software shutdown, bleach clean, DI rinse, and vent sheath tank.

Dye Selection Guidance

Guidance on picking dyes for optimal excitation with available lasers.

Dye Compatibility and Excitation

Dye Compatibility Table

List of dyes compatible with SH800 sorter's lasers and their excitation.

Laser Limitations and Recommendations

Note on no RED laser and sub-optimal excitation for certain dyes with yellow-green laser.