Quality assurance for polychromatic flow cytometry using a suite of calibration beads
Abstract
The quality assurance program presented here provides a means to maximize and maintain the performance of individual flow cytometers in a facility. To optimize performance, we recommend performing all three steps (optimization, calibration and standardization) in this program when a new flow cytometer is installed or whenever the flow cytometer's optical path is altered (e.g., lasers, filters or detectors are replaced). The complete process requires 3-4 h. On a more frequent basis, only a subset of these procedures need to be performed as a part of daily maintenance routines. The data generated can be tracked to monitor the instrument and determine whether service is needed. In addition, the data can provide a metric for whether repairs and upgrades have improved or harmed performance, and for future instrument-to-instrument comparisons. In sum, the procedures presented here represent an updated framework for optimizing, calibrating and standardizing a flow cytometer for daily use.
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Voltage titration is the most widely accepted approach to optimize detector voltages and determines the minimally acceptable voltage yielding optimal resolution for each detector.
“The most widely accepted approach for this is a voltage titration [3], which will determine the minimally acceptable voltage yielding optimal resolution for each detector.”
From Guidelines for the use of flow cytometry and cell sorting in immunological studies (third edition) (2021) · cites this paper as [3]
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Self-citationQuality assurance for polychromatic flow cytometry
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Guidelines for the use of flow cytometry and cell sorting in immunological studies (third edition)
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Human bone marrow assessment by single-cell RNA sequencing, mass cytometry, and flow cytometry
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- Authors
- Perfetto, S. P., Ambrozak, D., Nguyen, R., Chattopadhyay, P. K., & Roederer, M.
- Affiliations
- National Institutes of Health (US)
- Corresponding author
- Perfetto, Stephen P
- Venue
- Nature Protocols
- Published
- 2012-11-08
- Volume
- 7
- Issue
- 12
- Pages
- 2067-2079
- ISSN
- 1750-2799 (ISSN-L)
- 1754-2189
- OpenAlex
- W2034043217
- Cited by
- 148
- References
- 10
- Citations by year
- 2026: 6 · 2025: 10 · 2024: 10 · 2023: 15 · 2022: 14 · 2021: 15 · 2020: 12 · 2019: 16 · 2018: 8 · 2017: 13 · 2016: 10 · 2015: 8 · 2014: 6 · 2013: 4 · 2012: 1
- FWCI
- 4.08
- Type
- journal article
- Language
- en
- Open access
- closed
- Licence
- www.springer.com/tdm (tdm)
- Locations
- Nature Protocols (publishedVersion)
- PubMed (publishedVersion)
- Funders
- National Institutes of Health (Z99 AI999999)
- Topics
- Health Sciences › Medicine › Physiology › Clinical Laboratory Practices and Quality Control
- Health Sciences › Health Professions › Medical Laboratory Technology › Quality and Safety in Healthcare
- Social Sciences › Decision Sciences › Statistics, Probability and Uncertainty › Scientific Measurement and Uncertainty Evaluation
- Keywords
- polychromatic flow cytometry, instrument optimization, instrument maintenance, performance monitoring
- MeSH terms
- Calibration, Flow Cytometry, Fluorescent Dyes, Quality Control
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