The Gladstone Flow Cytometry Core enables scientists to analyze or isolate cells based on their phenotypic or functional properties, and can be used in conjunction with other core technologies to form a robust and effective pipeline for cellular analysis. Analysis of up to 18 and sorting of up to 15 fluorescent parameters is available from a wide variety of samples and cell types. Our staff is experienced in experimental and antibody panel design, practical operation of multiple flow cytometric platforms, and basic and advanced data analysis.

Contact

Jane Srivastava
Core Director

 

Services Provided

  • Cell sorting under sterile conditions with BSL-2 or BSL-3 containment for blood-borne pathogens.
  • Cell sorting (up to four ways and 15 fluorescent parameters) and/or analysis (up to 18 fluorescent parameters) for extracellular and intracellular markers as well as fluorescent proteins.
  • Single cell sorting into 96-well plates for cloning or sequencing.
  • High-throughput analysis from 96- and 384-well plates (up to 18 fluorescent parameters).
  • Functional marker sorting and analysis (e.g., phospho-flow, cell cycle, and mitochondrial assays).
  • Image cytometry using the Amnis ImageStream.

Core Members

Michael Daniel
Student Intern

Nandhini Raman
Senior Research Technologist

Jane Srivastava
Research Scientist

FAQs

How Do I Book a Session on a Flow Cytometer or Cell Sorter?

Contact core staff if you have not used the core before. You will need to make an account on iLab before beginning any work.

Which Flow Cytometers Are Capable of Running Plates?

The Fortessa X-20 and Attune NxT are capable of analyzing 96 and 384 well plates.

What Does Cytometer Training Involve?

You will receive a 1.5-hour instruction on how to operate the cytometer and use the software. You will also receive instruction on basic troubleshooting and access to online resources.

What Type of Samples Can Be Run on the Cytometer?

Any single-cell suspension where cells are between 500nm and 20um can be run. Samples can be provided in 5ml Falcon tubes, 15ml Falcon tubes, Eppendorf tubes, multi-well plates, or slides.

Cells can be at a concentration of up to 5 million per mL in PBS or media, but serum should be limited to 2 percent serum, as more than this can affect the charge applied to the stream. You can also add HEPES (25mM), EDTA (1mM) or DNAse (10U/ml) to avoid clumping. If you are concerned that your sample is too concentrated, bring some media to dilute your sample in. Filter your cells through a cell strainer or filter-top tube.

Can the Cytometers Be Used Independently, after Hours, or on Weekends?

After being trained, you must demonstrate proficiency in cytometer operation, maintenance, and basic troubleshooting before being allowed to use the equipment independently or outside of operating hours.

Does the Flow Core Archive Data?

The flow core saves a copy of the FCS files onto an external hard drive.

Capabilities

 

Single cell sorting enables scientists to effectively isolate an individual cell for downstream processes such as cloning and RNAseq. Index sorting of single cells provides the location on an analysis plot of each cell from each well.

Cell cycle analysis in flow cytometry entails the binding of certain dyes such as Propidium Iodide or Hoechst to DNAr base pairs after permeabilization. This enables sorting of cells based on whether they are in G0/G1, S or G2/M phase. Analysis programs help to determine the accurate percentages of cells in each of these phases.

 
 

Multi-parameter immunophenotyping has many applications including disease detection, development and tracking. Optimal fluorescent antibody panel design can help distinguish rare and/or novel cell populations.

Fees and Scheduling

For more information on our fees, contact Jane Srivastava or Nandhini Raman.

Publications

A molecular mechanism for probabilistic bet hedging and its role in viral latency. Sonali Chaturvedi, Jonathan Klein, Noam Vardi, Cynthia Bolovan-Fritts, Marie Wolf, Kelvin Du, Luwanika Mlera, Meredith Calvert, Nathaniel J. Moorman, Felicia Goodrum, Bo Huang, Leor S. Weinberger.

HIV efficiently infects T cells from the endometrium and remodels them to promote systemic viral spread. Tongcui Ma, Xiaoyu Luo, Ashley F George, Gourab Mukherjee, Nandini Sen, Trimble L Spitzer, Linda C Giudice, Warner C Greene, Nadia R Roan.

SIRT1 is downregulated by autophagy in senescence and ageing. Caiyue Xu, Lu Wang, Parinaz Fozouni, Gry Evjen, Vemika Chandra, Jing Jiang, Congcong Lu, Michael Nicastri, Corey Bretz, Jeffrey D. Winkler, Ravi Amaravadi, Benjamin A. Garcia, Peter D. Adams, Melanie Ott, Wei Tong, Terje Johansen, Zhixun Dou, Shelley L. Berger.

Remote Training of SRL Users and Staff in a Global Pandemic. Kathleen Daniels, Alexis Conway, Rui Gardner, Lola Martinez, Kylie M. Price, Sarah Schneider, Rachael Sheridan, Jane Srivastava, Sherry Thornton.

Shared Resource Laboratory Operations: Changes Made During Initial Global COVID‐19 Lockdown of 2020. Jessica B. Back, Cora H. Chadick, Juan J. Garcia Vallejo, Eva Orlowski‐Oliver, Radhika Patel, Caroline E. Roe, Jane Srivastava, Rachael V. Walker.

Transcription Factor Overexpression Drives Reliable Differentiation of Retinal Pigment Epithelium from Human Induced Pluripotent Stem Cells. Tessa E. Dewell, Ketrin Gjoni, Angela Z. Liu, Ashley R.G. Libby, Anthony T. Moore, Po-Lin So, Bruce R. Conklin.

Shared Mechanisms Govern HIV Transcriptional Suppression in Circulating CD103+ and Gut CD4+ T Cells. Steven A. Yukl, Shahzada Khan, Tsui-Hua Chen, Martin Trapecar, Frank Wu, Guorui Xie, Sushama Telwatte, Daniel Fulop, Alexander R. Pico, Gregory M. Laird, Kristen D. Ritter, Norman G. Jones, Chuanyi M. Lu, Robert F. Siliciano, Nadia R. Roan, Jeffrey M. Milush, Ma Somsouk, Steven G. Deeks, Peter W. Hunt, Shomyseh Sanjabi.

FOXO1 promotes HIV Latency by suppressing ER stress in T cells. Albert Vallejo-Gracia, Irene P. Chen, Rosalba Perrone, Emilie Besnard, Daniela Boehm, Emilie Battivelli, Tugsan Tezil, Karsten Krey, Kyle A. Raymond, Philip A. Hull, Marius Walter, Ireneusz Habrylo, Andrew Cruz, Steven Deeks, Satish Pillai, Eric Verdin, Melanie Ott.

Silencing of E-cadherin in induced human pluripotent stem cells promotes extraembryonic fates accompanying multilineage differentiation. Ashley RG Libby, Ivana Vasic, David A Joy, Martina Z Krakora, Fredrico N Mendoza-Camacho, Bruce R Conklin, Todd C McDevitt.

In Vivo Chimeric Alzheimer’s Disease Modeling of Apolipoprotein E4 Toxicity in Human Neurons. Ramsey Najm, Kelly A.Zalocusky, Misha Zilberter, Seo Yeon Yoon, Yanxia Hao, Nicole Koutsodendris, MaxineNelson, Antara Rao, Alice Taubes, Emily A. Jones, Yadong Huang.

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