schematic of multiple fluorescence spectra showing emission overlap

Controls, compensation and calibration are all critical to obtaining accurate results in flow cytometric analysis. In this section you will find educational resources including application notes, videos, articles and other useful tools to help you set up your flow cytometer and analyze your samples.

Controls, compensation and calibration features


graphic of 3-color detection compensation in flow cytometry

Basics of flow cytometry, Part II: Compensation

This presentation provides an overview of basic fluorochromes used in flow cytometry including the principle of compensation and how it is performed as well as the types of controls recommended and their use.

Scientific poster

two-panel graph showing PMT response

Evaluation of multiple techniques for PMT optimization (PDF)

This study compares a variety of techniques and calculations for setting your flow cytometer photomultiplier tube and obtaining the minimum voltage required.

Controls, compensation and calibration learning resources

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Application note (2018) High-throughput automation with the Attune NxT Autosampler: consistent results across all wells and across plates Attune/Attune NxT Flow Cytometer, Autosampler
Application note (2015)Attune NxT Flow Cytometer for 6-color immunophenotyping analysis of stained human whole blood using a no-lyse, no-wash protocol, with no compensationAttune NxT, flow cytometer, immunophenotyping, multicolor flow cytometry
Application note (2015) 13-parameter immunophenotyping of human lysed whole blood with the Attune NxT Flow Cytometer allows for the identification of B cells, NK cells, multiple T cell subsets and myeloid cells Alexa Fluor, antibodies, Attune NxT, compensation, flow cytometer, fluorescent dyes, immunophenotyping, multicolor flow cytometry, nanocrystals, Qdot
BioProbes article (2018)A comprehensive resource for state-of-the-art flow cytometry methods. Guidelines for the use of flow cytometry and cell sorting in immunological studiesantibodies and proteins, cell counting, cell cycle, cell proliferation, flow cytometer calibration, flow cytometry, flow cytometry compensation, flow cytometry sample preparation, immunology, immunophenotyping, viability
BioProbes article (2015)Journal Club—Multiparameter detection of early apoptosis markers without compensationapoptosis, Attune NxT, cell structure-mitochondria, flow cytometry compensation, fluorescence, immunophenotyping
BioProbes article (2015)No-compensation immunophenotyping using a four-laser flow cytometerAttune NxT, flow cytometer, fluorescence, immunophenotyping, multicolor flow cytometry
BioProbes article (2014)Obtain the highest-quality data your system can deliver—Fluorescent microspheres for calibrating microscopes and flow cytometerscalibration, cell counting, flow cytometer, fluorescence microscopy/fluorescence imaging, microscope calibration, particles
BioProbes article (2009) Accurate compensation is as easy as AbC calibration, compensation, fluorescent dyes, immunophenotyping, multicolor flow cytometry, particles
Molecular Probes HandbookMicrospheres—Section 6.5flow cytometer calibration, fluorescence microscopy/fluorescence imaging, microscope calibration, particles
Molecular Probes HandbookFlow cytometry reference standards—Section 23.2flow cytometer
ProtocolsOneComp and UltraComp Compensation Beads protocols for flow cytometry (Invitrogen eBioscience reagents)compensation, flow cytometry
Scientific poster (2018) Maintaining even temperature distribution over chilled samples in 96-well plates and micro-centrifuge tubes academic research solutions, microplates, quality control, temperature control
Scientific poster (2018) A comparison of GFP BrightComp eBeads Compensation Bead Kit performance against multiple GFP variants Attune/Attune NxT, beads, flow cytometry compensation, fluorescent protein detection,
Scientific poster (2018) Evaluation of multiple techniques for PMT optimization Attune/Attune NxT Flow Cytometer, flow cytometer calibration, photomultiplier settings
Scientific Poster (2016) A microsphere-based universal compensation approachAttune NxT, Flow cytometer calibration, flow cytometry sample preparation
Scientific poster (2010) Use of surface-modified microspheres for setting compensation in multicolor flow cytometry antibodies, cells, flow cytometry compensation, fluorescent dyes, multicolor flow cytometry, particles, violet laser-excited reagents
Tutorial videoMolecular Probes Tutorial Series—Analyzing Flow Cytometry Data flow cytometer calibration, flow cytometry
Tutorial videoMolecular Probes Tutorial Series—Introduction to Flow Cytometry flow cytometer calibration, multicolor flow cytometry
WebinarBasics of flow cytometry, Part II: Compensation
This presentation provides an overview of basic fluorochromes used in flow cytometry. Topic includes: the principle of compensation, how to perform compensation, the types of controls recommended and their use, basic strategies for designing a flow experiment, and data presentation.
flow cytometry data, flow cytometry set up, multicolor flow cytometry
WebinarBasics of multi color flow cytometry panel design
With the proliferation of new fluorescent dyes, as well as instruments that can detect 18 or more parameters multicolor flow cytometry has become more popular and more accessible than ever. This webinar presented by Dr. Holden T. Maecker at Stanford University will discuss the caveats of good panel design, including:
  • Rules for designing panels
  • Examples and practical application of these rules
  • Controls and standardization
  • Relevance of panel design to new mass cytometry platforms
antibodies, compensation, multicolor flow cytometry