Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

Flow Cytometry01:23

Flow Cytometry

The development of flow cytometry techniques began in 1934 with initial attempts by Andrew Moldavan, a bacteriologist who counted the cells in a flowing capillary system. Moldavan pumped cells through a capillary tube focused under a microscope for visualization. The invention of photometry allowed the measurement of differentially-stained cells, and Louis Kamentsky developed the first multiparameter flow cytometer in 1965 to identify and count the cancer cells in cervical tissue specimens.
In...

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Identification of CD24 as a marker of Patched1 deleted medulloblastoma-initiating neural progenitor cells.

PloS one·2019
Same author

Functional domain analysis of SOX18 transcription factor using a single-chain variable fragment-based approach.

mAbs·2018
Same author

Targeting membrane proteins for antibody discovery using phage display.

Scientific reports·2016
Same author

Purification of neural precursor cells reveals the presence of distinct, stimulus-specific subpopulations of quiescent precursors in the adult mouse hippocampus.

The Journal of neuroscience : the official journal of the Society for Neuroscience·2015
Same author

Development of a novel cell sorting method that samples population diversity in flow cytometry.

Cytometry. Part A : the journal of the International Society for Analytical Cytology·2015
Same author

A novel flow cytometric method to assess inflammasome formation.

Journal of immunology (Baltimore, Md. : 1950)·2014

Related Experiment Video

Updated: Jun 4, 2026

Kinetic Measurement and Real Time Visualization of Somatic Reprogramming
08:56

Kinetic Measurement and Real Time Visualization of Somatic Reprogramming

Published on: July 30, 2016

A method of quantifying cell sorting yield in "real time".

Geoffrey W Osborne1

  • 1Flow Cytometry Laboratory, Queensland Brain Institute, University of Queensland, Brisbane, QLD 4072, Australia. g.osborne@uq.edu.au

Cytometry. Part a : the Journal of the International Society for Analytical Cytology
|February 4, 2011
PubMed
Summary

Calibration beads used in cell sorting flow cytometers overestimate cell yield. This study shows that cells have lower sorting yields than indicated by beads, revealing trends related to nozzle tip diameter and improving real-time yield assessment.

More Related Videos

In Situ Microscopy for Real-time Determination of Single-cell Morphology in Bioprocesses
07:26

In Situ Microscopy for Real-time Determination of Single-cell Morphology in Bioprocesses

Published on: December 5, 2019

Immunophenotyping and Cell Sorting of Human MKs from Human Primary Sources or Differentiated In Vitro from Hematopoietic Progenitors
14:30

Immunophenotyping and Cell Sorting of Human MKs from Human Primary Sources or Differentiated In Vitro from Hematopoietic Progenitors

Published on: August 7, 2021

Related Experiment Videos

Last Updated: Jun 4, 2026

Kinetic Measurement and Real Time Visualization of Somatic Reprogramming
08:56

Kinetic Measurement and Real Time Visualization of Somatic Reprogramming

Published on: July 30, 2016

In Situ Microscopy for Real-time Determination of Single-cell Morphology in Bioprocesses
07:26

In Situ Microscopy for Real-time Determination of Single-cell Morphology in Bioprocesses

Published on: December 5, 2019

Immunophenotyping and Cell Sorting of Human MKs from Human Primary Sources or Differentiated In Vitro from Hematopoietic Progenitors
14:30

Immunophenotyping and Cell Sorting of Human MKs from Human Primary Sources or Differentiated In Vitro from Hematopoietic Progenitors

Published on: August 7, 2021

Area of Science:

  • Biotechnology
  • Cell Biology
  • Analytical Chemistry

Background:

  • Cell sorting flow cytometers rely on calibration beads to determine optimal drop delay times for accurate cell sorting.
  • The assumption that beads accurately predict cell sorting yield is central to current automated technologies.

Purpose of the Study:

  • To investigate the accuracy of calibration beads in predicting actual cell sorting yields.
  • To develop a method for real-time, accurate quantification of cell sorting yield.

Main Methods:

  • HEK293T cells were incubated with Accudrop™ calibration beads, and bead incorporation was confirmed via imaging cytometry.
  • Cells were sorted using commercially available cell sorters with automated drop delay calibration.
  • Sorting experiments were conducted to compare bead-indicated yield with actual cell recovery.

Main Results:

  • Cell sorting yields were consistently lower than those indicated by calibration beads.
  • A clear correlation was observed between nozzle tip diameter and sorting yield.
  • A novel method for on-the-fly yield quantification was demonstrated, independent of drop charge counts.

Conclusions:

  • Calibration beads may overestimate cell sorting efficiency, impacting experimental recovery expectations.
  • Nozzle tip diameter is a significant factor influencing cell sorting yield.
  • The developed method offers a more accurate approach to assessing cell sorting yield in real-time.