34th Congress of the International Society for Advancement of Cytometry

Sunday, June 23, 2019


Creating a Repository of Best-Practice Guidelines for Successful Cell Sorting Outcomes

Gelo dela Cruz
Peter Lopez
Anna Fossum

This workshop will focus on the different variables involved in cell sorting that SRL professionals deem important to document and collect to determine successful cell sorting outcomes. Also, a consensus from attendees will be obtained on how to determine successful sorting outcomes. The gathered information will serve as basis for a database/repository that will be available as a reference for live cell sorting experiments.

I.        Scope and Limitation of the Workshop
II.       Results of Pre-congress Survey
III.     Determining Successful Cell Sorting Discussion (based on survey)
IV.     Variables involved in Cell Sorting Discussion (based on survey)
V.      Summary

Pre-Conference Survey for Creating a Repository of Best-Practice Guidelines for Successful Cell Sorting Outcomes can be found at https://goo.gl/forms/xmPpCnIJN2KGgNTp1

Strategies for Successful Conflict Resolution: You Can't Hide in Your Office and Hope the Problem Goes Away

Jessica Back
Ann Marie Deslauriers-Cox

This workshop will be based on customer service with an emphasis on creating effective working relationships with disgruntled or challenging customers. Strategies that may be used to help resolve workplace conflicts will be presented. Using real-world examples submitted by the SRL community, participants will discuss ways to handle difficult situations.

The format will include slides with real-world scenarios, interactive participation, and live polling. A pre-conference survey will be sent out asking CYTO attendees to describe conflicts they have dealt with and indicate how these were handled, whether they felt the resolution was appropriate, and what they would have done differently. Any common conflict themes identified in the survey will be incorporated into the workshop discussion by the facilitators.

15-20 minute guided discussion with audience participation and feedback on various techniques for dealing with conflict.
1. Polling on conflict resolution techniques
2. Group discussion of most popular methods

15-20 minute presentation of real-world scenarios with audience participation on how to resolve the presented scenario followed by information on how it was actually handled.
1. Presentation of scenario
2. Audience input
3. What occurred? What was successful?
4. Other techniques which could be useful in the future.

15-20 minute small-group breakout to discuss current issues attendees may be dealing with and input from peers on methods for resolving these issues.

Pre-conference Survey Questions
  1. Describe a time when you had a conflict with a colleague.
  2. How was the conflict resolved?
  3. If given the chance, what would you have done differently to resolve this conflict? 
Pre-Conference Survey link: https://www.surveymonkey.com/r/8SQ7T5S

The Status of Flow Cytometry in Africa

Facilitators: Lydia Tesfa, Alfonso Blanco, Bill Telford, Iyadh Douagi 
For the survey associated with this workshop, click here

To date, numerous experts in cytometry field, in particular those involved in various projects in Africa, have put emphasis on providing more resources to African scientists and clinicians involved in cytometry related studies. To assess the level of expertise and all other related issues in the cytometry field, survey questions have been sent out targeting the African Scientific Community. Therefore, the main objective of the workshop is to present survey results and open up a round table discussion on emphasis on key problem areas identified by the survey.

Key questions: 
Discuss key problem areas as identified by the survey; 
How do we secure sustainable resources moving forward? Can we approach Melinda & Bill Gates Foundation and other potential donors; 
How do we secure full support from vendors? How do we make them more engaged? 
Can we choose a site in Africa, to have a workshop every 2 or 3 years?
Can we find ways to persuade instrument companies to give instrument based workshops in Africa?
How do we strengthen ties between African scientific community with ISAC society?

- Welcome & Introduction & Objectives

- Session 1
    1. Speaker 1: Present summarized survey results (Lydia + Alfonso)
    2. Speaker 2 : J. Paul Robinson
    3. Speaker 3: Bill Telford
    4: Speaker 4: Iyadh Douagi
- Session 2
    1. Break out into sub groups to brainstorm ideas
    2. Presentation of summarized results

- Session 3
    1. Conclusions and summarizing outcomes of the workshop (as a group)

Applying Scatter & Fluorescence Standardization to Flow Cytometric Data of Small Particles

Facilitators: Joshua Welsh, Vera Tang

Standardization methods are not well utilized and instrument calibration is often not performed or reported for small particle analysis. It is critical for the development of the field that all flow cytometry measurements are reported in standardized units to allow for comparisons to be made between platforms with different sensitivities. In this workshop we will go over some proposed methods for scatter and fluorescence calibration using scatter modeling and fluorescence standards to better report EV data.


1. Introduction to the benefits of fluorescence standardisation using MESF beads and scattering standardisation using NIST traceable beads with scatter modelling software, highlighting the MIFlowCyt standards, flow repository and the EV minimum considerations for performing flow cytometry (15 mins). 

2. Interactive demonstrations of performing linear regression and converting axes to MESF units using test dataset from FlowRepository (15 mins). There will be intervals throughout the protocol for discussions on advantages and limitations of the standardisation method and future improvements.

3. Interactive demonstration using scatter modelling software with test dataset from FlowRepository (15 mins). There will be intervals throughout the protocol for discussions on advantages and limitations of the standardisation method and future improvements.

4. Concluding remarks and open discussion for the methods demonstrated and future methods or materials that need to be developed to advance the field. (15 mins)

Resources can be found at www.joshuawelsh.co.uk/cyto2019

How to Ensure Robustness and Reproducibility when Using Mass Cytometry for Clinical Studies?

Jose Estevam
Diana L. Bonilla Escobar
Holden Maecker
Adeeb Rahman 
Jonathan Irish
James Lederer 
Brice Gaudillière
Nima Aghaeepour
Elena Hsieh Professor
Greg Behbehani 
Mike Leipold
Radhika Rayanki
Peter Krutzik
Chris Groves

Mass cytometry is a cutting-edge technology that merges the high-throughput single cell analysis with the specificity and resolution of mass spectrometry. It allows over forty parameters to be measured simultaneously on single cells, allowing deeper profiling of samples. While mass cytometry has highly contributed to basic scientific discovery, some of the major challenges come when trying to implement this technology in clinical studies. The addition of mass cytometry to the pool of technologies in clinical studies definitely requires more standardization and quality control. Discussion is needed on the validation of CyTOF assays for clinical use including application validation, reagents and protocols, instrument operation, quality control and data analysis.  We hope to bring together all the experts in the field to learn from their experience on how to overcome these challenges.

1. Reagent and Protocol Optimization
2. Panel Validation
3. Assay Validation
4. Instrument Validation and Quality Control
5. Data Analysis and Statistical Evaluation
6. Quality Control
7. Regulation

Monday, June 24, 2019

User Training in Cell Sorting in Core Facilities – Eliminating the Staff to Instrument Bottleneck

Christina Ewald
Derek Davies
Claudia Dumrese

Staff and instrument availability limits the access to cell sorters for untrained researchers, which in turn limits the scope of sorting experiments, which are realistically possible per instrument. Training end users as well as dedicated staff operators in cell sorting is a solution favoured by only a handful of facilities up to now. This workshop will assess risks, costs and benefits, outline the required training effort, staff resources, infrastructure including instrumentation and aims to establish a roadmap to successfully introduce user sorter training programmes at interested institutions.

Detailed Agenda (60 min workshop):

Welcome and introduction of workshop schedule. 5 min
Presentation of participant pre-workshop survey results.
GAP analysis current sorting service and training practices.
Assessment of the need for a change in these in the community.

Formal presentation providing an overview of the challenge at hand as well as practical experience report from the sorter training programmes at the University of Zurich and the Francis Crick Institute in London. 10 min
Speakers: Derek Davies and Christina Ewald

Group work of participants on individual aspects guided and documented by facilitators. (3-4 groups) 25 min

The following aspects will be addressed in groups:
Group 1: Is a self-service model financially viable?
Group 2: What should a training look like? A one-suits-all vs. 3-will-suit-most training strategy consensus. Is a European, American, worldwide training network feasible?
Group 3: How is success of training judged?
Group 4: Positive and negative experiences, hopes & concerns, limitations, hurdles and how to address them (staff, financial, institutional, user base)
How can industry help the Cores?

Facilitator feedback, summary, communication of consensus. 15 min

Wrap-up, information of availability of documentation, access to Best Practice material and SOPs, where to find support in the future. 5 min

The outcomes of the survey and workshop discussions will be made available to ISAC members after the Congress.

Finding Solutions to Difficult Developmental Biology and Stem Cell Flow Cytometry Issues

Facilitator: Gelo dela Cruz

This workshop will focus on issues arising from the analysis and sorting in developmental biology and stem cell differentiation experiments. In these experiments, researchers are faced with difficulty in isolating rare samples from solid tissues and determining proper controls for analyzing in vitro development, among others. The workshop will rely on input from attendees with diverse sorting and analysis experience.

I.          Introduction to Workshop
          a.    Scope and Limitations
II.         Discussion
          a.    Isolation and analysis of rare cells from developing organs (pancreas, etc)
          b.    Separation and analysis of differentiating stem cells from feeders in culture
          c.    Controls for in vitro differentiation samples
          d.    Other issues from survey
III.        Summary

Pre-Conference survey for the workshop Finding Solutions to Difficult Developmental Biology and Stem Cell Flow Cytometry Issues can be found at https://goo.gl/forms/6gzIWKAku6JJkltw2

SRL Careers: What Can ISAC Do to Help Us?

Yasmin Hacque
Rachael Walker
Aja Rieger
Julfa Begum
Kylie Price

Within many SRLs there is a career track that leads toward becoming a core manager/core director.  This track however, does not exist in all institutes or companies.  Therefore, junior SRL members are often uncertain of how to move up the career ladder.  A key aim of this workshop is to discuss how to provide support for junior SRL staff or newly appointed SRL managers.  Conversely, this workshop aims to cover what the next career steps for an established core manager could be.

Workshop Agenda:
Time (mins):

5 Introduction

10 Survey Results: Survey designed to address the need for careers advice or mentorship for SRL staff members from across the field.

10  Present and discuss current options for getting on the career ladder.  An open forum on sli.do will be integrated into the discussion.

15 Group Discussions:
Each group will be given an anonymous case study to discuss.  The aim is for participants to draw from their own experiences.  A facilitator will be present in each group to take notes.

* Group 1: Case 1: New to an SRL:
SRL members who have recently embarked on a career in cytometry and are looking for guidance to further their career. 

* Group 2: Case 2: Newly appointed SRL manager:
These new managers are likely to be looking for different support from ISAC, in areas such as budgeting, hiring staff and line management.

* Group 3: Case 3: Established SRL manager/director:
These experienced SRL managers/directors maybe looking for a change in career direction and perhaps would like support and advice from other senior SRL members who have considered or successfully made a career change.

5 Group 1 Summary

5 Group 2 Summary

5 Group 3 Summary

5 Conclusion: Summary culminating in a proposed plan of action (for initiating any ideas/programs).

Total: 60 mins

Pre-Conference Survey: https://goo.gl/forms/hTC6Gxqfmquy66i02
For pre-conference pdf, click here.

When the Shoe Doesn't Fit: Defining and Overcoming Barriers to Achieving Best Practices for Solo-Staff SRLs

Roxana del Rio-Guerra
Laura Lewis-Tuffin

In 2016, a group of ISAC members published a comprehensive document establishing standards and guidance for seven best practices in Flow Cytometry Core Facility/Shared Resource Laboratories (SRLs) (Barksy et al. Cytometry Part A 89A:1017). These standards were intended to be achievable by any SRL. Within the SRL model, a considerable number operate with a single staff member who provides all services (solo-staff SRLs). In these SRLs, the low staff number can create special challenges to reaching best practice status in all seven areas. The major questions we want to address in this workshop are: 1. Which of the seven best practices are easy and which are difficult for solo-staff SRLs to implement? 2. What are common barriers to implementation of best practices in solo-staff SRLs? 3. What strategies are solo-staff SRLs using to breach some of the barriers?

1. Introduction, brief summary of the seven best practice areas and results of pre-meeting survey regarding the best practice areas most difficult for solo-staff SRLs to approach.
2. Small group analysis of best practice areas, with a focus on identification of barriers to implementation, and brainstorming workarounds to those barriers.
3. Summary presentations of small group discussion points for each best practice area.

Prior to the workshop, it is helpful for attendees to review either (a) or (b).
(a) Barsky L.W. et al. “International Society for Advancement of Cytometry (ISAC) Flow Cytometry Shared Resource Laboratory (SRL) Best Practices” 2016 Cytometry Part A 89A: 1017-1030.
(b) If member of ISAC, please go to cytou.org > Shared Resource Lab > SRL Best Practices Series

Attendees should also be on the lookout for a pre-meeting survey on this topic.

Flow Cytometry Data for Clinical Trials, Manual or Automated Analysis?

Michael Nathan Hedrick
Yongliang Steve Sun
Cherie Green
Ryan Brinkman

This workshop is intended for translational and clinical researchers from biotech/pharmaceutical industry, regulatory agencies, clinical laboratories, contract research organizations, and any flow cytometrists who want to generate high quality and reproducible data.
Flow cytometry data analysis is the last but not the least step to ensure high quality data. Manual (human driven) analysis is the current gold standard. However, manual analysis poses challenges such as capacity and rigor, and the subjectivity can introduce the largest variability in the assay. Automated analysis methods have been maturing over the last few years, with advantages of throughput, efficiency, consistency and reproducibility, and provide a solution to analyzing large sets of flow data from clinical studies. This workshop will focus on pros and cons of manual vs. automated analysis, and how to make a choice based on each individual project. The following questions (but not limited) will be discussed:
- How to decide on using manual or automated analysis (supervised and unsupervised gating)
- How to validate automated analysis
- New challenges and opportunities resulting from the use of automated analysis
- How to QC data generated from automated analysis
The most important aspect of this workshop is to open a dialog, and to generate ideas for how to use automated approaches to analyze flow cytometry data from clinical trials. The outcome will be an opinion/a white paper.

- Introduction and attendee survey, 5 min
- Presentation of case studies, 10 min
- Interactive discussions and recommendations regarding the above questions, 40 min
- Summary and concluding remarks, 5 min

Workshop 11
CYTO Lab Hacks: A Platform for the Exchange of Innovations in Cytometry

Jakub Nedbal 
Claudia Bispo
Christopher Hall
Bunny Cotleur

Our aim is to develop an accessible, free and open online platform centered around a dynamic cytometry innovation community to allow sustained collaboration and sharing of innovations and ideas. During the workshop, we will present several non-commercial cytometry innovations deposited and documented through various online repositories. The workshop audience will interactively test and provide feedback on the innovations, the repositories and documentation styles to guide the development of an online platform and guidelines for sharing innovations.

This workshop will be highly interactive, focusing on a moderated discussion with the aim to form a consensus on the following three areas:

* What makes an innovation generate interest and enthusiasm in the cytometry community?
* Which online platforms fared best as repositories of cytometry innovations?
* What are the minimum requirements for documentation to make the innovation useful and transferable to other labs?

We will actively engage with the cytometry community prior to the workshop. After the workshop, we will conduct an assessment meeting and publish draft guidelines and roadmap to develop CYTO Lab Hacks online repository for free and open to cytometry innovations.

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