Experimental Design

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Immunology:

This site, run by the British Society for Immunology, offers useful guidance for those who are not familiar with immunology.

Tips:

This page features a large collection of tips and tricks for a wide variety of flow cytometry applications, including cell sorting, cell cycle analysis, antibody titration, statistical analysis, and flow cytometry controls.

Cell counting:

The Chromocyte website provides a video from Stem Cell technologies on how to count cells using a haemocytometer, an integral preparatory step for many flow cytometry experiments.

Sample preparation:

Chromocyte offers two helpful resources on sample preparation. The first is an excyte list of simple mistakes to avoid in preparing for flow cytometry, and the second is a webinar from Miltenyi Biotech on sample preparation for flow cytometry.

Instrument and fluorophores:

This website module includes resources aimed for helping users understand their instrument, reagent brightness, expression of common markers, as well as the basic rules to effective panel design. 

Marker expression:

Under the “expression of common markers” item, this helpful page from BioLegend lists the expression levels of a wide variety of common surface receptors. Whilst only human antigens are listed, this is a very helpful first step in planning flow cytometry panels.

Brilliant Violet panel design:

This webinar features advice on planning panels incorporating the Brilliant Violet dyes, in the context of large polychromatic flow cytometry experiments.

Fluorophore combinations:

The chromocyte website offers a module with links to resources for understanding how to pick the best fluorophore combinations. This section doesn’t contain much primary knowledge, but contains a collection of links to other useful sites.

 

Flow cytometry controls:

This webinar describes the use of flow cytometry controls in the context of advanced data analysis. This webinar is free for ISAC members, but costs $37.50 USD for non-members.

Designing, planning, and performing polychromatic flow cytometry experiments:

CYTOU provides an excellent webinar from Pratip Chattopadhyay, describing the best practices in hardware optimization and reagent utilization, as well as the latest data analysis considerations. This webinar is free for ISAC members, but costs $37.50 USD for non-members.

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