Single‐Calibration Cell Size Measurement With Flow Cytometry

ABSTRACT Measuring the size of individual cells in high‐throughput experiments is often important in biomedical research and applications. Nevertheless, popular tools for high‐throughput single‐cell biology, such as flow cytometers, only offer proxies of a cell's size, typically reported in arb...

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Bibliographic Details
Published in:Cytometry. Part A Vol. 107; no. 4; pp. 263 - 270
Main Authors: Davies, Philip, Cavallaro, Massimo, Hebenstreit, Daniel
Format: Journal Article
Language:English
Published: Hoboken, USA John Wiley & Sons, Inc 01.04.2025
Wiley Subscription Services, Inc
Subjects:
Animals
Calibration
Cell Size
computational biology
Flow
Flow cytometry
Flow Cytometry - instrumentation
Flow Cytometry - methods
Flow Cytometry - standards
Humans
Lasers
Linear Models
Linear transformations
Medical research
Parameters
Scattering
Single-Cell Analysis - methods
Size determination
computational biology
flow cytometry
calibration
cell size
ISSN:1552-4922, 1552-4930, 1552-4930
Online Access:Get full text
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Summary:ABSTRACT Measuring the size of individual cells in high‐throughput experiments is often important in biomedical research and applications. Nevertheless, popular tools for high‐throughput single‐cell biology, such as flow cytometers, only offer proxies of a cell's size, typically reported in arbitrary scales and often subject to changes in the instrument's settings as selected by multiple users. In this paper, we demonstrate that it is possible to calibrate flowcytometry laser scatter signals with accurate measures of cell diameter from separate devices and that the calibration can be conserved upon changes in the laser settings. We demonstrate our approach based on flow cytometric sorting of cells of a mammalian cell line according to a selection of scatter parameters, followed by cell size determination with a Coulter counter. A straightforward procedure is presented that relates the flow cytometric scatter parameters to the absolute size measurements using linear models, along with a linear transformation that converts between different instrument settings on the flow cytometer. Our method makes it possible to record on a flow cytometer a cell's size in absolute units and correlate it with other features that are recorded in parallel in the fluorescence detection channels.
Bibliography:Philip Davies and Massimo Cavallaro contributed equally to this work.
This work was supported by EPSRC grant EP/T002794/1 and BBSRC grant BB/M017982/1.
Funding
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ISSN:1552-4922
1552-4930
1552-4930
DOI:10.1002/cyto.a.24924