Indirect immunofluorescence microscopy in cultured cells

S P Wheatley1, Y L Wang

  • 1Cell Biology Group, Worcester Foundation for Biomedical Research, Shrewsbury, Massachusetts 01545, USA.

Insights

Learn about indirect immunofluorescence, a powerful technique for visualizing cellular targets. This guide offers protocols and considerations for researchers new to fluorescence immunolocalization methods.

Area of Science:

  • Cell Biology
  • Immunology
  • Microscopy

Background:

  • Fluorescence immunolocalization has advanced significantly since the 1960s.
  • Innovations in probe chemistry, microscopy, and image detection have enhanced the technique.
  • Indirect immunofluorescence is a widely used method in biological research.

Purpose of the Study:

  • To provide an overview of the current status of indirect immunofluorescence.
  • To serve as an entry point for researchers new to the technique.
  • To cover general considerations from cell culture to image detection.

Main Methods:

  • Indirect immunofluorescence techniques.
  • Cell culture protocols.
  • Image detection strategies.
  • Probe chemistry considerations.

Main Results:

  • The chapter details general considerations for applying indirect immunofluorescence.
  • It includes specific protocols for initiating use of the technique.
  • The overview covers the entire workflow from sample preparation to image acquisition.

Conclusions:

  • Indirect immunofluorescence is an evolving and valuable technique.
  • This chapter provides a practical guide for new users.
  • The presented protocols facilitate the adoption of fluorescence immunolocalization.