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Introduction to Flow Cytometry
Rating: 4.3 out of 5(90 ratings)
265 students
Created byLavinia Ionescu
Last updated 9/2022
English

What you'll learn

  • Understand the fundamental principles of flow cytometry
  • Identify the components of a flow cytometer and describe their role
  • Interpret instrument quality control data
  • Understand how to determine what fluorochromes can be detected by a flow cytometer based on fluorochrome spectra and instrument configuration
  • Summarize characteristics of fluorochromes that are highly relevant in practice (single/tandem, brightness, fluorescence spillover and spreading potential)
  • Gain a working understanding of fluorescence compensation
  • Plan a flow cytometry experiment using adequate controls
  • Learn common first steps for troubleshooting

Course content

5 sections21 lectures2h 15m total length
  • Introduction2:48

    An overview of the course.

  • What Is Cytometry and What Does It Do?9:58

    A definition of flow cytometry, brief historical summary and fundamentals of light scattering. Students will be able to describe the significance of forward and side scatter (FS/FSC and SS/SSC).

  • Fluorescence Overview4:14

    Definitions of fluorescence and related terms.

  • Excitation and Emission Spectra6:01

    Explanation of fluorochrome excitation and emission spectra. These concepts are highly relevant for fluorochrome (dye) choices, in conjunction with the capabilities of the flow cytometer(s) being used.

  • Spectral characteristics of phycoerythrin (PE)

Requirements

  • No flow cytometry experience needed.

Description

⭐Average of ALL reviews received, equally weighted: 4.2 stars.⭐

This course is designed to help beginner or occasional users of flow cytometry become more familiar with the numerous facets of the field, starting with a theoretical perspective. It may also be useful to intermediate or more frequent operators as a review of the concepts that remain firmly in place across platforms: from the basics of light scattering and fluorescence to the universal main components of flow cytometers, the practical principles of multicolor compensation, and the roles of experimental controls typically used in flow cytometry.

Please feel welcome to check out the curriculum/descriptions of the individual videos for a few additional details that may offer a more accurate idea of the scope of the course.

VERY IMPORTANT; PLEASE NOTE:

  • The course is NOT focused on any specific software or instruments and is NOT intended to substitute hands-on training and/or certification courses. Instead, it aims to present the elements that constitute a shared ground among platforms and provide a springboard to advanced flow cytometry concepts.

  • The course does NOT go into highly specific details regarding sample preparation, acquisition and analysis. This is due to the variety of possible applications, instruments, reagents and dedicated software, as well as the wide online availability of such protocols (often tailored to particular commercial products) for the most commonly used assays - e.g., cell phenotyping using surface vs. intracellular markers, counting, apoptosis, cell cycle, proliferation, "exotic applications" etc. The instructor is not affiliated with any commercial entity and does not endorse any particular commercially available product (instrument, reagent, software or other) in this course.

The instructor has 15 years of experience in flow cytometry, at first in academia as a biomedical researcher, then in the industry as a flow cytometry application scientist.

The lectures are built with the awareness that flow cytometry is used for numerous applications, almost all of which rely upon detection of fluorescent molecules. Moreover, in today's "flow" landscape, one-laser instruments coexist with multi-laser analyzers capable of detecting dozens of colors, and several powerful options are available in terms of analysis software.

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Captions: The course has captions written from scratch (10 of 21 videos), as well as automatic captions - kindly provided by the platform - that have been reviewed and edited (where necessary) by the instructor.

Who this course is for:

  • Beginner or occasional users of flow cytometry.