Uploaded July 2021 | Updated September 2026, 2 weeks ago
Join us for a chat with Bio-Rad application scientists about the current issues most relevant to your research. Jump to specific topics by following the links below.
In this episode, we discuss: Cell Sorting Tips for Success
- Essentials for sample preparation
- Caring for your cells during the sorting process
- Controls, viability dyes, and other factors to consider while sorting
- Choosing flow cytometry experimental controls
We had so many great audience questions. Watch the whole video or quickly jump to a specific question:
00:10 Introduction
3:38 What is the best method for cell dissociation, cell scraping or trypsin?
5:03 What is the best way to overcome or prevent clogging? I get clogs with some of my samples.
9:00 My sorts aren't as pure as I need them, I'm only getting 60% of my target cells, what can I do to improve?
11:10 Any tips for preventing contamination of sorted cells?
13:10 Should you add viability dye again to the sample you are re-running to test purity?
15:10 General challenge with sorting that I experience is the yield beyond the purity that has been mentioned. How can one increase sorted cell yield beyond increasing the initial starting material?
19:40 Any suggestions for what quality of water to use when making 1x sheath buffer from 10x buffer?
20:50 Every time I sort into plates my cells die. Why is this?
22:20 What temperature should I sort my cells at, and is it OK to sort into 10% FBS?
24:40 Sorting speed is a challenge, how can one strike the balance because some cell sorting takes very long?
25:08 I keep getting contamination in my sorts, how can I better prevent this?
31:10 How fast can I sort? Does speed really matter?
34:25 What percent Contrad do you use for cleaning?
34:55 What's the best way to clean a nozzle if I have a clog?
36:15 How robust are fresh blood cells during sorting? For multiple cell sorting is it better to do this as ones with multiple staining or step-wise?
38:40 I'm trying to sort primary mouse hepatocytes based off GFP but I'm not getting enough and keep clogging the instrument, can you recommend anything?
42:00 Is there anything I need to take into consideration when sorting bacteria, that is different from mammalian cells?
44:05 What is a good, live/dead for bacterial sorts?
45:55 I am getting a loss of my target receptor over time while my sample is sitting in the sorter. What can I do to stop this? i.e. my positive population is decreasing.
48:45 I think my sort is going well, but when I do my post sort purity run, I am getting really low numbers for yield compared to sample input. Do you have any suggestions for why this might happen?
50:22 Is it okay to run samples reconstituted in Media containing FBS (MPP media) as compared to only 1X DPBS?
51:55 My post sort viability is low (40%) and I really need it higher for my downstream application. I think my cells might not like being sorted, what can I do?
55:45 It's late, I want to go home. How important is it to follow the shutdown & startup procedures?
57:40 When performing single-cell sorting, on highest purity mode, filtered iPSc cells, without viability dye due to toxicity, and one still observes several cells in a couple of wells in a 96 well plate, how could this be improved or avoided?
59:35 Conclusion
Attend a live coffee chat episode by visiting bio-rad.com/coffeechat
Subscribe to the Bio-Rad channel: youtube.com/user/BioRadLifeScience?sub_confirmation=1
We Are Bio-Rad.
Our mission: To provide useful, high-quality products and services that advance scientific discovery and improve healthcare. At Bio-Rad, we are united behind this effort. These two objectives are the driving force behind every decision we make, from developing innovative ideas to building global solutions that help solve our customers' greatest challenges.
Connect with Bio-Rad Online:
Website: bio-rad.com
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©2021 Bio-Rad is a trademark of Bio-Rad Laboratories, Inc. in certain jurisdictions. All trademarks used herein are the property of their respective owner.
Music used by permission granted by royalty-free license from Pond5.com
Join us for a chat with Bio-Rad application scientists about the current issues most relevant to your research. Jump to specific topics by following the links below.
In this episode, we discuss: Cell Sorting Tips for Success
- Essentials for sample preparation
- Caring for your cells during the sorting process
- Controls, viability dyes, and other factors to consider while sorting
- Choosing flow cytometry experimental controls
We had so many great audience questions. Watch the whole video or quickly jump to a specific question:
00:10 Introduction
3:38 What is the best method for cell dissociation, cell scraping or trypsin?
5:03 What is the best way to overcome or prevent clogging? I get clogs with some of my samples.
9:00 My sorts aren't as pure as I need them, I'm only getting 60% of my target cells, what can I do to improve?
11:10 Any tips for preventing contamination of sorted cells?
13:10 Should you add viability dye again to the sample you are re-running to test purity?
15:10 General challenge with sorting that I experience is the yield beyond the purity that has been mentioned. How can one increase sorted cell yield beyond increasing the initial starting material?
19:40 Any suggestions for what quality of water to use when making 1x sheath buffer from 10x buffer?
20:50 Every time I sort into plates my cells die. Why is this?
22:20 What temperature should I sort my cells at, and is it OK to sort into 10% FBS?
24:40 Sorting speed is a challenge, how can one strike the balance because some cell sorting takes very long?
25:08 I keep getting contamination in my sorts, how can I better prevent this?
31:10 How fast can I sort? Does speed really matter?
34:25 What percent Contrad do you use for cleaning?
34:55 What's the best way to clean a nozzle if I have a clog?
36:15 How robust are fresh blood cells during sorting? For multiple cell sorting is it better to do this as ones with multiple staining or step-wise?
38:40 I'm trying to sort primary mouse hepatocytes based off GFP but I'm not getting enough and keep clogging the instrument, can you recommend anything?
42:00 Is there anything I need to take into consideration when sorting bacteria, that is different from mammalian cells?
44:05 What is a good, live/dead for bacterial sorts?
45:55 I am getting a loss of my target receptor over time while my sample is sitting in the sorter. What can I do to stop this? i.e. my positive population is decreasing.
48:45 I think my sort is going well, but when I do my post sort purity run, I am getting really low numbers for yield compared to sample input. Do you have any suggestions for why this might happen?
50:22 Is it okay to run samples reconstituted in Media containing FBS (MPP media) as compared to only 1X DPBS?
51:55 My post sort viability is low (40%) and I really need it higher for my downstream application. I think my cells might not like being sorted, what can I do?
55:45 It's late, I want to go home. How important is it to follow the shutdown & startup procedures?
57:40 When performing single-cell sorting, on highest purity mode, filtered iPSc cells, without viability dye due to toxicity, and one still observes several cells in a couple of wells in a 96 well plate, how could this be improved or avoided?
59:35 Conclusion
Attend a live coffee chat episode by visiting bio-rad.com/coffeechat
Subscribe to the Bio-Rad channel: youtube.com/user/BioRadLifeScience?sub_confirmation=1
We Are Bio-Rad.
Our mission: To provide useful, high-quality products and services that advance scientific discovery and improve healthcare. At Bio-Rad, we are united behind this effort. These two objectives are the driving force behind every decision we make, from developing innovative ideas to building global solutions that help solve our customers' greatest challenges.
Connect with Bio-Rad Online:
Website: bio-rad.com
LinkedIn: linkedin.com/company/1613226
Facebook: facebook.com/biorad
Twitter: twitter.com/BioRadLifeSci
Instagram: @BioRadLabs
Snapchat: @BioRadLabs
©2021 Bio-Rad is a trademark of Bio-Rad Laboratories, Inc. in certain jurisdictions. All trademarks used herein are the property of their respective owner.
Music used by permission granted by royalty-free license from Pond5.com
