00:35:21 Sandesh Rimal: Good evening everyone 😊 00:35:44 Shova Shrestha: Namaskar & Good evening Everyone. 00:35:51 Bivaan Bajracharya: Good evening 00:36:31 Desh: Namate and Good evening!! 00:37:37 Gorkha Raj Giri: Namaste 00:41:11 Tara Shrestha: Namaste and good evening everyone 00:43:01 Purshotam Kumar Sah Kanu: Namaste and Good Evening everyone 00:49:06 Soma Kanta Baral: Namaste and good evening everyone 00:54:15 William Telford: https://isac-net.org/general/custom.asp?page=CYTO-University 01:00:55 William Telford: CYTO 2021 conference information at https://www.cytoconference.org 01:06:35 Dr Krishna Sharma: Hello from Bhutan to everyone in the class. 01:09:30 William Telford: That's wonderful, thank you for joining us! 01:21:05 William Telford: Feel free to chat in questions! 01:21:36 Pragati Pradhan: Sure sir. 01:26:24 Krishna Manandhar: Great Zosia! We are hearing perfectly and clear. Nice going ….. 01:28:02 Himal Chhetri: Dear Professor, I wanna know whether it can detect nonviable cells or only viable cells. 01:55:20 William Telford: You can detect both non-viable and viable cells, and use cytometry techniques to measure the number and proportion of both types. We can measure apoptosis (directed cell death) using many reagents including DNA binding dyes and annexin V. 01:56:09 William Telford: You can also distinguish non-viable and viable cells by forward and side light scatter, although you should also include a viability reagent. 01:56:39 William Telford: We can talk about apoptosis assays after Zosia's talk and on Thursday. 02:03:44 William Telford: Apoptosis assays can be used to measure the effects of a drug. Apoptosis is a frequent readout for drug effects. 02:04:21 William Telford: Annexin V and a cell permeability probe (such as PI, a DNA dye) are the most common assay for apoptosis. Easy and relatively cheap compared to others. 02:04:54 Himal Chhetri: Oh, if so what's the possibility of nonspecific binding of dead cells to antibodies? And that might gives false positive results. 02:04:56 William Telford: But you can also just use PI (propidium iodide) alone as a preliminary screen to see if you drug is inducing cell death. 02:05:17 Purshotam Kumar Sah Kanu: Wow, that's great 02:09:59 Himal Chhetri: Yeah. 7 - AAD viability dye might be equally useful. Thanks Prof. William Telford. 02:15:08 Aashish Dhakal: Dear professor, I have a query. My question is :- 02:15:14 Aashish Dhakal: are there primary targets for the isotype control on the cells during flow cytometry and how can we know this? 02:16:20 William Telford: Yes, 7-AAD is also a good viability dye - it appears in the 488 nm laser long red detector on most cytometers. It combines well with FITC annexin V. 02:17:32 William Telford: Isotype control antibodies are used to assess non-specific binding of labeled antibodies. So we use them in place of our specific antibody to see the contribution of non-specific antibody binding to total fluorescence. 02:18:35 William Telford: However, they have a habit of binding in unpredictable ways. Sometimes they can attach to unknown markers. 02:19:06 Aashish Dhakal: Thank you Professor William 🙏 02:19:18 William Telford: For this reason, they are not so commonly used anymore, except in situations where the risk of non-specific binding is very high (like intracellular cytokine flow).. 02:20:00 William Telford: Full minus one (FMO) controls are now more common - labeled with everything except one antibody, to see the contribution of all other labels to background of that label/detector. 02:21:23 Tinmaya Rai: hello, what was bdscameans 02:22:14 Charu Arjyal (Tri Chandra Campus, Kathmandu): th. 02:23:04 Charu Arjyal (Tri Chandra Campus, Kathmandu): Thank you so much for the nice presentation, Zosia and the organizers for the opportunity! 02:24:50 shova shrestha: Thank you Zosia for wonderful presentation. 02:25:05 Jarina Joshi: In PM Detector, higher voltage is better or should be optimized 02:29:50 Dipendra Kumar Mandal: will you please tell us, which dye is used for red fluorescence? 02:31:17 Jarina Joshi: Thank you. 02:33:01 Dipendra Kumar Mandal: thank you 02:33:03 Sugam Pokharel: Thanks T. William and Zosia 02:33:12 Zosia Maciorowski: off red laser APC, Alexa647, Alexa 700, APC tandems, live dead dyes 02:33:27 Arun kumar Thakur: which method is best for research( single panel ,optics or electronics)? 02:35:46 Upasana Shrestha: Thank you. 02:35:54 Amulya Baral: Thank You