January 2022: Michael Kissner/CSCI Flow Cytometry Core

Background:

What is the main focus of your lab?

We are a core facility that provides comprehensive and cutting-edge flow cytometry services to CSCI investigators as well as the Columbia University research community at large. Our services have two focuses: first, we provide access to high-end and well-maintained flow cytometry instrumentation, including cell sorters, cell analyzers, an imaging cytometer and other supporting instrumentation (Luminex technology and qPCR). Our model focuses on self-service, independent operation of all instruments, including the cell sorters, for maximum accessibility and flexibility to accommodate all users’ experiments. Our second focus is to provide support services to ensure users generate the highest quality data held to high standards of rigor and reproducibility. We have developed a comprehensive instrument training program for independent usage, and we offer theoretical training through classroom-based instructions. Additionally, we support users’ experiments through experimental design, data analysis, and troubleshooting consultations.

We are also very excited about our newest service, beginning in early 2022, integrating a 10X Genomics platform for a seamless cells-to-cDNA workflow.

How long have you had your lab? When did you join Columbia University?

I have been at Columbia since January 2017. I joined Columbia and CSCI with Dr. Passegué and helped established the flow cytometry core from the ground up.

How big is your lab currently?

We have three team members: myself and two cytometrists: Rose Gordon-Schneider and Jangho Lee.

Where is your lab located?

We are located in the CSCI main space on the 11th floor of the William Black Building.

Current affairs:

What are the most exciting projects/directions in the lab at this moment?

While we don’t directly perform research, we have extensive exposure to many labs’ exciting projects through the collaborative process of supporting users’ flow cytometry needs. Perhaps unique to the CSCI Flow Cytometry core facility, we strive to establish scientific relationships with all of our users. We aim to be familiar with each user’s research needs to be sure that we are providing the most appropriate services. Many of these relationships take on a collaborative nature, which allows us to be a part of exciting, cutting-edge, and impactful research. These interactions have allowed us to develop our skills applying flow cytometry, especially cell sorting, of a wide array of sample types including neurons, tumor, pancreas, gut, iPSC-derived tissues of various types, and nuclei among many others. In particular, we have developed expertise in fine-tuning sorting protocols and instrument setup for high viability, purity, and recovery, especially of difficult and delicate sample types. We have applied these skills to samples with downstream scRNAseq applications through the 10X platform and our users consistently generate excellent sequencing data from cells sorted on our instruments.

One very exciting upcoming project is the integration of the 10X Genomics Chromium scRNAseq workflow to the core facility. This service will be implemented in partnership with the Columbia Genome Center and will allow users to seamlessly go from cells to cDNA within the same physical space. A key aspect of this service is that it will be user run. We have developed a comprehensive training process, including both post-sort cell processing and the 10X workflow, that will prepare users with the expertise to independently and competently complete the entire 10X Genomics protocol for 3’ or 5’ scRNAseq chemistry. Users will then be able to sort and create cDNA from their sample without being tied to staff availability. This will allow researchers to sequence unpredictable samples without restrictions.

The entire workflow will be supported, including instrumentation for the intermediate step of cell counting and viability assessment, and a dedicated space and associated equipment (thermocycler, centrifuge) will be provided for the 10X Genomics workflow. The Genome Center will help supply reagents for the 10X workflow and provide downstream sequencing services. This is a very unique partnership that can be a model for the ever more important relationship between cell sorting and sequencing.

One type of technology that the core is currently lacking is full-spectrum flow cytometry. This type of cutting-edge detection technology uses full-spectrum detection of every fluorophore individually among up to 184 photodetectors and unmixing algorithms to measure up to 45+ fluorochromes in the same sample. There are several key benefits of full-spectrum flow cytometry. First, because the measurement of each fluorochrome is not restricted to a single detector, fluorochrome choice is much broader than it is in conventional flow cytometry. Second, dyes that are too close to be resolved by conventional flow cytometry can be easily resolved by full-spectrum detection. As such, many more colors can be used simultaneously, allowing establishment of true high-parameter panels. Finally, an autofluorescence spectrum can be generated and unmixed along with the fluorochromes, effectively lowering autofluorescence signal, which can be a major benefit for certain problematic samples.

Full-spectrum flow cytometry is becoming an essential technique and is required for more and more of our users. As such, we are planning on writing an S10 shared instrument grant for the cutting-edge Sony ID7000 spectral analyzer, which is the preeminent state-of-the-art spectral analyzer, equipped with up to 7 lasers and 184 photodetectors. We are very eager to be able to provide access to full-spectrum cytometry to our users.

What are the biggest accomplishments that your lab recently had?

The past year has been one of very exciting changes for the core facility.

We added the NovoCyte Quanteon flow cytometer analyzer in early 2021, and the machine has been a big hit among our users. The Quanteon is a 4-laser, 27-parameter machine with automated plate and tube sampling. One of the reasons we picked this instrument is because of its stellar and easy-to-use software, which has really streamlined user training. It has been so successful that we are planning on adding a second unit to meet users’ needs.

Additionally, we recently acquired a new sorter, the Sony MA900, which is a highly-automated, extremely user-friendly instrument that has really taken off. It’s been running daily since installation, and users are very excited about its capabilities and ease-of-use. In particular, training for independent operation is significantly more streamlined that is for the FACSAria, and we have found that one or two 2-hour training sessions, depending on the user’s experimental needs, is sufficient for full competence on all aspects of normal operation. The MA900 has been so successful that we are acquiring a second unit to meet our significant cell sorting needs.

Training:

What's your best approach to mentoring trainees in the lab?

Although staff in a core facility play a different role than trainees in a research lab, there are some similarities, depending on the mentoring style. For me, the most important objective for a core staff member is career growth. I think it’s misleading to think of core facility staff as technicians or simply instrument operators. I want the staff in our lab to feel empowered to take a leading role in all aspects of the user-facing side of operations and to lead collaborative efforts with users they have formed relationships with. Although turnover in a core facility can be problematic due to the amount of time it takes to become skilled in flow cytometry - at least 1-2 years - I want our staff to grow and ultimately pursue the career path in flow cytometry that most interests them. Our last team member, Daniel Troast, is now in a fantastic role at a leading instrument manufacturer, and his training here in CSCI got him there. He had no experience in flow cytometry before joining and is now supporting highly complex instrumentation. That type of legacy is an important metric of the success of a facility’s ability to mentor and train staff. I think it would be fantastic if every staff member who has worked with us ended up managing their own core facility!

Who were your most influential mentors/role models in science and what did you learn from them?

Emmanuelle has been an incredible mentor for me in my endeavor to start and sustain the core facility. Her leadership and vision for the core facility and CSCI in general has truly shaped what I have been able to accomplish at Columbia.

A big part of this has to do with the organization culture in CSCI. Last summer, I was invited by the Office for Research, along with Barbara Corneo, to participate in the Leadership and Management in Core Facility course offered by Northwestern University's Kellogg School of Business. The course was taught by business school professors and was a very unique opportunity to explore topics that we don’t really think about much - organization culture, leadership, teams, and marketing and budgeting strategies. One of the most compelling sections of the course was focused on organization culture. One of the ways to define culture is a sense of aligned goals, principles, and practices shared amongst members of an organization. After thinking about this, I realized that large academic institutions in general, comprised of so many entities, struggle with maintaining a common culture across the whole organization. I then realized that one of the things I really love about CSCI is how cohesive the community is. A big part of this is due to the fact that Emmanuelle’s leadership and vision for CSCI has nurtured a culture where members feel included, supported, and part of its community. I think this is a very special aspect of CSCI!

In the flow cytometry field as well, community is very important. We rely on each other to share ideas and experiences and to troubleshoot. I am active in the local flow cytometry communities and find a great deal of mentorship among more experienced colleagues who manage other core facilities in the area.