Practical Guide to Sony SH800 Sorter

Tip: If only analysis is performed (without sorting), click on Analyzer mode to shorten the calibration time required.

Calibration will automatically execute the four steps listed on the left side of the screen. After completing everything, check if the result shows' completed successfully '.

If successful, click OK to enter the "Create Experiment" window.

Chapter 7: Experiment Creation and Parameter Setting

After calibration is passed, a new experiment can be created.

7.1 Basic Information Input

Fill in information such as experiment name, operator, cell type, etc. in Experiment Information.

Enter the marker name and select the corresponding fluorescent dye.

7.2 Collection Parameters and Laser Selection

Select the parameter types to be collected: Area, Height, and Width, which are used for FSC, BSC (lateral scattering), and each fluorescence channel (FL1~FL6), respectively.

If not all lasers are selected at startup, you can add the required laser wavelength selection here.

After completing the setup, click on Create Experiment, and the main window will display the Experiment Explorer.

7.3 Template based Creation

Users can also create new experiments based on existing experiment templates, or directly copy recently used experiments as templates.

Chapter 8: Compensation Settings and Threshold Adjustment

Before starting to collect samples, it is usually necessary to complete fluorescence compensation and adjust detection sensitivity.

8.1 Automatic Compensation Wizard

Go to the Compensation tab and click on the Compensation Wizard to start the automatic compensation process.

Follow the guide prompts to load each single staining tube and negative tube in sequence.

After compensation is completed, the software will automatically generate a compensation matrix and apply it to the current experiment.

8.2 Detector Threshold Adjustment

Click on Detector&Threshold in the middle left corner of the screen.

Adjust the sensitivity values of each detector until a clear event cluster appears on the scatter plot.

Set appropriate thresholds (usually using FSC or fluorescence thresholds) to eliminate debris and noise.

Important reminder: Each sample tube must be filtered with a blue filter cartridge before loading. Microfluidic chips are extremely sensitive to blockage, and any particle with a diameter larger than the nozzle size will cause complete blockage.

Chapter 9: Precautions for Experimental Collection and Sorting

Confirm that the sample tube has been filtered and placed on the correct tube holder.

Before sorting, it is recommended to use non sorting mode (Analyzer mode) to observe the sample status and adjust the gate strategy.

Choose the appropriate sorting mode (purity first, yield first, or single-cell mode) during sorting.

If using a porous plate for single-cell sorting, the index sorting function can be enabled for easy traceability in the future.

Chapter 10: Shutdown and Cleaning - Irreconcilable Steps

Errors or skipping shutdown cleaning are the most common causes of permanent damage to sample lines. The following order must be strictly followed.

10.1 Start shutdown program

Select the Cytometer tab and click on [Software and Device] in the Shutdown menu.

Never click Shutdown directly in the pop-up shutdown window! Directly clicking will immediately turn off the instrument power and skip the cleaning step, which will cause the samples in the sample line to dry up and become blocked, which cannot be repaired. Users need to purchase a new sample line at their own expense.

You must click Start to start the cleaning process.

10.2 Cleaning steps

Prepare a 15 ml Falcon tube and fill it with 10 ml FACS Clean solution (bleach containing hypochlorite). Load using the corresponding sample tube holder. Attention: Do not overfill with bleach.

Software guidance completes bleach cleaning.

After cleaning, remove the bleach tube and click Next.

Prepare another 15 ml Falcon tube and fill it with 12-13 ml of ultrapure water (MilliQ). The level of DI water must be higher than the previous level of bleach to ensure that all bleach on the sample probe is thoroughly rinsed off.

Perform DI water rinse.

10.3 Turn off the instrument

After cleaning, the software asks if the host and software will automatically shut down. If using a standard sample line (not self owned), click Shutdown.

If using your own sample line and/or chip, you should cancel the shutdown and log out from the current account to return to the login interface. Then contact the facility staff to assist in retrieving the sample line and chip.

Shut down the computer.

Turn off the compression pump.

Release all pressure inside the sheath fluid tank: Lift the circular pressure relief valve at the top until no more gas is released. Long term storage under pressure may damage the instrument.

Emptying the waste liquid tank: Disconnect the pink waste liquid pipe, carefully unscrew the lid (to prevent liquid from flowing into the air filter), and pour the waste liquid into the sink. Wipe off any splashing waste liquid and disinfect with 70% ethanol. Reconnect the waste liquid tank.

Chapter 11: Common Problems and Troubleshooting