BD FACSCelesta Flow Cytometer User Manual (English)

Questions and Answers:

J
Jessica BarkerAug 19, 2025
What to do if I see droplets on the SIP of my BD FACSCelesta Flow Cytometer Measuring Instruments?
- M
  Michael WallaceAug 19, 2025
  If you observe droplets on the SIP of your BD Measuring Instruments, several factors could be responsible: * A worn O-ring in the retainer: Replace the O-ring. * The outer sleeve not seated in the retainer: Loosen the retainer, push the outer sleeve up until seated, then tighten the retainer. * The outer sleeve not on the sample injection tube: Loosen the retainer, slide the outer sleeve over the sample injection tube until seated, and tighten the retainer. * A pinched waste line: Check the waste line. * A full waste tank: Empty the waste tank. * The droplet containment vacuum not functioning: Call your BD service representative. * The mode is not set to HTS: Change the mode to HTS by pressing down the MODE button for more than 3 seconds.
R
reynoldssusanAug 23, 2025
How to fix high event rate on BD FACSCelesta Flow Cytometer Measuring Instruments?
- J
  Jonathan LeeAug 25, 2025
  To address a high event rate on your BD Measuring Instruments: * Remove air bubbles from the sheath filter or flow cell. * Increase the threshold level. * Set the PMT voltage lower for the threshold parameter. * Dilute the sample if it is too concentrated. * Set the sample flow rate to MED or LOW.
C
Colleen CaseyAug 26, 2025
What causes erratic event rate in BD FACSCelesta Flow Cytometer and how to resolve it?
- R
  Rebecca HessAug 26, 2025
  An erratic event rate in BD Measuring Instruments can stem from several causes. Here's how to address them: * If the sample tube is cracked, replace it. * If there's air or debris in the flow cell, prime the fluidics system. * If the Bal seal is worn, replace it. * If the sample injection tube is clogged, remove the sample tube to allow backflushing; if the issue persists, clean the sample injection tube. * If the sample is contaminated, prepare the specimen again, ensuring the tube is clean. * If the sheath filter is dirty, replace it.
M
Michael BrownAug 28, 2025
How to resolve distorted scatter parameters on BD Measuring Instruments?
- M
  Maria RamirezAug 28, 2025
  To resolve distorted scatter parameters on BD Measuring Instruments: * Optimize the scatter parameters. * Purge air from the sheath filter. * Flush the system if the flow cell is dirty. * Ensure the sheath container lid is tight and all connectors are secure to prevent air leaks. * Replace hypertonic buffers or fixative.
K
Kevin MilesSep 1, 2025
What causes excessive debris in display on BD FACSCelesta Flow Cytometer and how to fix it?
- J
  Julia GeorgeSep 1, 2025
  If you observe an excessive amount of debris in the display of your BD Measuring Instruments, here's what could be happening and how to address it: * Threshold level is too low: Increase the threshold level. * Sheath filter is dirty: Replace the filter. * Flow cell is dirty: Flush the system. * Dead cells or debris in the sample: Examine the sample under a microscope. * Sample is contaminated: Re-stain the sample, ensuring that the tube is clean. * Stock sheath fluid is contaminated: Rinse the sheath container with DI water, then fill the container with sheath fluid from another (or new lot) bulk container.
H
Henry HallSep 3, 2025
Why is my sample tube not fitting on SIP of BD FACSCelesta Flow Cytometer?
- S
  Scott PriceSep 4, 2025
  If your sample tube isn't fitting on the SIP of your BD Measuring Instruments, it could be due to: * Using sample tubes other than Falcon® tubes: Use Falcon 12 x 75-mm polystyrene sample tubes. * A worn Bal seal: Replace the Bal seal. * A cracked sample tube: Transfer contents to a new tube.
D
Deanna ChapmanSep 5, 2025
What to do if there are no events in acquisition display and RUN button is green on my BD Measuring Instruments?
- J
  Jennifer ThomasSep 5, 2025
  If there are no events in the acquisition display and the RUN button is green on your BD Measuring Instruments, check the following: * The threshold is not set to the correct parameter: Set the threshold to the correct parameter for your application. * The threshold level is too high: Lower the threshold level. * The PMT voltage is set too low: Set the PMT voltage higher for the threshold parameter. * There may be a gating issue: See the BD FACSDiva Software Reference Manual for information on setting gates. * There may be air in the sheath filter: Purge the filter.
D
Douglas GarciaSep 7, 2025
How to fix low event rate on BD Measuring Instruments?
- A
  Alyssa KelleySep 7, 2025
  To address a low event rate on your BD Measuring Instruments: * Lower the threshold level. * Prime the fluidics system to remove any air bubbles or debris in the flow cell. * Set the PMT voltage higher for the threshold parameter.
J
Jeremy MunozSep 9, 2025
Why is there no fluorescence signal on my BD FACSCelesta Flow Cytometer?
- T
  Tracy BoltonSep 9, 2025
  If you are not getting a fluorescence signal on your BD Measuring Instruments, it could be due to: * Incorrect fluorochrome assignment: Make sure that the cytometer configuration in the software matches the optical filters in the cytometer and the configuration is as expected. * Laser malfunction: Call your BD service representative.

BD FACSCelesta Flow Cytometer Specifications

General

Type	Flow Cytometer
Data Acquisition Rate	Up to 35, 000 events per second
Software	BD FACSDiva Software
Laser Configuration	blue (488 nm), red (640 nm), violet (405 nm)
Sample Volume	Variable sample volume

Summary

About this guide

What this guide covers

Describes the procedures necessary to operate and maintain the BD FACSCelesta flow cytometer and software features.

Conventions

Lists the safety symbols used in this guide to alert users to potential hazards.

About the BD FACSCelesta documentation

Describes the documentation available with the BD FACSCelesta flow cytometer, including publication formats and help system.

Instrument technical support

Provides information on how to contact BD Biosciences customer support for technical assistance.

Introduction

System overview

Provides an overview of the BD FACSCelesta system, including the cytometer, software, and optional components.

Cytometer overview

Describes the BD FACSCelesta flow cytometer as an air-cooled multi-laser benchtop instrument.

Control panel

Details the components and functions of the cytometer's control panel, including system indicators and buttons.

Fluidics system

Explains how the fluidics system carries the sample to the interrogation region and its system indicators.

Sheath and waste containers

Describes the sheath and waste containers, their capacities, and alarm conditions.

Optics

Describes the optical components of the BD FACSCelesta flow cytometer, including detector arrays and laser options.

Workstation

Describes the components of the BD FACSCelesta workstation, including the PC, software, and monitors.

Cytometer setup

Starting the cytometer and computer

Provides step-by-step instructions for powering on and starting the BD FACSCelesta cytometer and associated computer.

Preparing the sheath container

Details the procedure for preparing the sheath container, including checking fluid levels and connections.

Removing air bubbles

Explains how to remove trapped air bubbles from the sheath filter and sheath line to ensure accurate data.

Preparing the waste container

Describes how to prepare the waste container, including preventing overflow and handling biohazardous materials.

Priming the fluidics

Details the procedure for priming the fluidics system to remove air bubbles and debris from the flow cell.

About the optical filters and mirrors

Provides a description of the optical filters and mirrors used in the BD FACSCelesta detector arrays.

Custom configurations and baselines

Explains how to create custom configurations and define baselines for performance checks.

Maintenance

Maintenance overview

Provides an overview of routine maintenance and cleaning procedures for the BD FACSCelesta flow cytometer.

Cleaning the fluidics

Details the daily fluidics cleaning procedure to prevent clogging and remove residual dyes.

Shutting down the cytometer

Provides instructions on how to properly shut down the BD FACSCelesta cytometer, including performing daily cleaning.

Flushing the system

Describes how to perform an overall fluidics cleaning to remove debris and contaminants from system tubing.

Replacing the waste container cap

Details the monthly procedure for replacing the waste container cap to maintain system integrity.

Changing the sheath filter

Explains how to change the sheath filter, recommended every six months for optimal performance.

Changing the Bal seal

Provides instructions on how to replace the Bal seal, which ensures proper sample tube pressurization.

Changing the sample tube O-ring

Details the procedure for replacing the sample tube O-ring to ensure proper droplet containment vacuum function.

Cleaning or replacing the sheath gasket

Describes how to clean or replace the gasket on the sheath tank lid when needed.

Optimizing cytometer settings

Cytometer settings workflow

Outlines the workflow for optimizing cytometer settings using Cytometer Setup and Tracking and Application Settings.

Verifying the configuration and user preferences

Explains how to verify the cytometer configuration and user preferences before creating an experiment.

Running a performance check

Details how to run a performance check as part of quality control to monitor instrument performance.

Setting up an experiment

Guides users through creating a new experiment, specifying parameters, and adding compensation tubes.

Creating application settings

Describes how to create application settings for consistent and reproducible use of cytometer settings.

Recording compensation controls

Explains how to create and record compensation controls using the Compensation Setup feature.

Calculating compensation

Provides instructions on how to calculate compensation after recording data for single-stained controls.

Recording and analyzing data

Data recording and analysis workflow

Outlines the basic acquisition and analysis tasks using BD FACSDiva software for recording and analyzing data.

Preparing the workspace

Details how to prepare the workspace and apply application settings to an experiment before recording data.

Recording data

Provides an example of how to preview and record data for multiple samples on the cytometer.

Analyzing data

Explains how to analyze recorded tubes by creating plots, gates, and statistics views on a new worksheet.

Reusing an analysis

Describes how to use global worksheets to apply the same analysis to a series of recorded tubes.

Technical overview

About fluidics

Describes the BD FACSCelesta flow cytometer fluidics system, including hydrodynamic focusing.

About optics

Describes the optics system, including lasers, optical filters, and detectors, and their functions.

About electronics

Describes the electronics in the BD FACSCelesta flow cytometer, including pulse processing and digital data handling.

Troubleshooting

Cytometer troubleshooting

Lists possible problems and recommended solutions for common BD FACSCelesta flow cytometer issues.

Electronics troubleshooting

Provides possible causes and recommended solutions for BD FACSCelesta electronics-related issues.

Detector array configurations

Fluorescence spectra

Shows sample emission spectra from common fluorochromes and laser excitation lines for multicolor panel design.

About configuration maps

Describes the filter and mirror arrangements in the detector arrays for different configurations.

About the configuration

Details the configuration options available for the BD FACSCelesta flow cytometer, supporting up to three lasers.

Base configuration polygon maps

Describes how filters and mirrors are arranged for standard polygon configurations.

Supplies and consumables

Ordering information

Provides contact information for ordering spare parts and consumables from BD Biosciences.

Beads

Lists the available QC and CS&T beads, including particle type, laser, supplier, and catalog number.

Reagents

Lists available reagents such as sheath fluid, cleaning solutions, and antibodies with their catalog numbers.

Equipment

Lists essential equipment items like Bal seal, O-ring, and sheath filter assembly with supplier and catalog numbers.

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