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 Experiment Videos

Fluorescent erythrocyte ghosts as standards for quantitative flow cytometry

S K Doberstein1, G Wiegand, L M Machesky

  • 1Department of Cell Biology and Anatomy, Johns Hopkins University, School of Medicine, Baltimore, Maryland, USA.

Cytometry
|May 1, 1995
PubMed
Summary
This summary is machine-generated.

Related Concept Videos

You might also read

Related Articles

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

Sort by
Same author

A RAC-GEF network critical for early intestinal tumourigenesis.

Nature communications·2021
Same author

Development of a cost-effective automated platform to produce human liver spheroids for basic and applied research.

Biofabrication·2020
Same author

Acquired von Willebrand syndrome in congenital heart disease surgery: results from an observational case-series.

Journal of thrombosis and haemostasis : JTH·2018
Same author

Pitfalls in Echocardiography: Coarctation of the Aorta Presenting as Dilated Cardiomyopathy (DCM).

Ultraschall in der Medizin (Stuttgart, Germany : 1980)·2015
Same author

Actin cytoskeletal control during epithelial to mesenchymal transition: focus on the pancreas and intestinal tract.

British journal of cancer·2015
Same author

Severe chest pain caused by Mycoplasma myocarditis in an adolescent patient.

Klinische Padiatrie·2013

We developed a fast, affordable method using erythrocyte ghosts to create flow cytometry standards with known fluorochrome content. These standards offer reliable fluorescence calibration for accurate cell analysis.

Area of Science:

  • Biotechnology
  • Analytical Chemistry
  • Cell Biology

Background:

  • Accurate calibration of flow cytometry fluorescence signals is crucial for quantitative cell analysis.
  • Existing standards, such as microbeads, can be expensive and may not cover all desired fluorochromes or concentrations.

Purpose of the Study:

  • To develop a quick, inexpensive, and versatile method for preparing flow cytometry standards with known fluorochrome content.
  • To characterize the performance of these novel standards for fluorescence calibration and quantitation.

Main Methods:

  • Erythrocyte ghosts were prepared via hypotonic lysis and filled with solutions of fluorescently labeled dextran.
  • The fluorescence intensity, linearity, particle volume, and stability of the ghost standards were evaluated.

Related Experiment Videos

  • Comparison of ghost standards with commercially available microbead standards.

    • Main Results:

    • The developed ghost standards exhibit a narrow fluorescence range and linear response up to 2 x 10(6) fluorochrome molecules/cell.
    • Ghost standard particles have a consistent volume of approximately 70 femtoliters/cell.
    • Fluorescence characteristics of ghost standards closely match those of comparable microbead standards.
    • Stable fluorescence was observed for at least 3 weeks at 4°C.
    • The method allows for the incorporation of a wide range of fluorochromes and concentrations.

    Conclusions:

    • This erythrocyte ghost-based method provides a cost-effective and rapid approach for generating reliable flow cytometry standards.
    • The versatility in fluorochrome incorporation and stability makes these standards valuable for various quantitative fluorescence applications.
    • This technique offers a viable alternative to existing commercial standards for flow cytometry calibration.