The Basics of Flow Cytometry |
Thank you for joining us on the Bio-protocol Ambassador Roundtable webinar on The Basics of Flow Cytometry with Mr. Derek Davies.
Explore the timestamps below to find exactly what you are looking for in the video!
⏰ Timestamps
0:00 Introduction or Overview
0:36 Definition of Flow Cytometry
2:25 Types of Flow Cytometers
2:55 Overview: Fluorescence Microscopy
3:38 Overview: Flow Cytometry
4:55 What does Flow Cytometry data look like?
6:13 Commercially available analysers
7:02 Components of a cytometer
8:30 Fluorescence and Fluorochromes
9:08 Fluorescence: Intrinsic and Extrinsic
11:27 Fluorescence: Physical Principles
13:05 Laser wavelengths
14:23 Fluorescence spectrum
16:40 Multiplexing fluorochromes
18:00 Types of optical filters (Long, short, band pass)
20:00 Fluorescence: Summary
20:36 Fluorochrome: Classes 1 and 2 (when to use which type?)
26:25 Fluorochrome: Brightness
27:48 How does a flow cytometer work? ~Components in detail
28:10 Sheath fluid
29:18 The flow cell: Hydrodynamic focusing
31:06 Fluorescence detection: Scattering of light, filters, detectors
37:58 How do we detect 'real' events? Concept of Threshold
40:48 How to represent the acquired data?
43:13 Fluorescence Compensation
47:14 Applications of flow cytometry (e.g. cell phenotyping, cell cycle, DNA analysis, proliferation assay, apoptosis, cytokine staining)
58:28 Summary: things to consider while designing your flow cytometry experiment
Q&A Session
1:00:00 Phosphorylated protein study, Gating strategies
1:01:40 Preparation, Storage and transportation of flow cytometry samples
1:03:16 Identifying a 'dirty' flow cytometer and procedure for cleaning of flow cytometer before and after the experiment
1:04:35 Use of experimental controls for flow cytometry experiment
1:05:55 Difference between and need of Compensation and FMOs
1:07:30 Difference between Spectral flow cytometer and conventional flow cytometer
1:09:35 How to navigate flow cytometry experiments as a beginner
1:10:44 Utilities and consumables for a flow cytometer
1:12:05 Scope of flow cytometry in vaccine studies
1:12:54 On handling limited biological samples in flow cytometry experiments and the minimum number of events needed to be considered
1:14:12 Closing remarks
🥼About:
Bio-protocol is an online peer-reviewed protocol journal curating and hosting high quality, free access, step-by-step protocols across the life sciences. Our motto is to improve research reproducibility and through our ambassador program for young researchers, we discuss openly about protocols, ideas and cutting-edge life science technologies.
~This webinar recording is on Flow Cytometry
Speaker: Derek Davies, Cytometry Consultant
Mr Davies has been a leader in flow cytometry for over 30 years now. He directed the Flow Cytometry Facility at what was the Imperial Cancer Research Fund in central London. Later, he became the head of the facility, then the Training lead of the Science technology Platform at the Francis Crick Institute, London. Over the years, he has organized and taught several flow cytometry training courses in the UK and beyond. He has been a chair of the flow cytometry section of the Royal Microscopical Society, chair of flowcytometry UK and has served on the council for the International Society for the Advancement of Cytometry. He now serves as a Cytometry Consultant; and organizes and shares information regarding upcoming training courses and development in Flow cytometry.
Connect with him @DDCytometry | linkedin.com/in/derek-davies-cytometry
Organizers:
Meet the Bio-protocol Ambassadors at https://bio-protocol.org/en/ambassado...
Dr. Antara Sengupta, CSIR- Institute of Genomics and Integrative Biology
Temitayo Bamgbose, Louisiana State University
Dr. Jack Hopkins, University College London
Priscilla Del Valle, UT Southwestern Medical Center
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