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
Email

 

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

Nandhini Raman
Senior Research Technologist

Jane Srivastava
Core Director

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.

More Publications

Support your research needs with our cutting-edge equipment and expertise.

Fortessa X-20

The Fortessa X-20 cell analyzer delivers high-performance, multicolor analysis and is configured with five lasers to detect up to 20 parameters (18 fluorescent and 2 scatter) simultaneously. An integrated High Throughput Sampler (HTS) provides rapid, fully automated sample acquisition from 96- and 384-well microtiter plates.

Practical and theoretical training is available for the Fortessa X-20, as well as experimental and data analysis support.

LASER CHANNEL BANDPASS FILTER LONGPASS MIRROR
UV (355 nm) BUV737 740/35 690LP
BUV496 515/30 450LP
BUV396 379/28  
Violet (405 nm) BV786 780/60 750LP
BV711 710/50 690LP
BV650 670/30 635LP
BV605 610/20 600LP
BV510/AMCYAN 470/15 450LP
BV421/BFP/DAPI 431/28 410LP
Blue (488 nm) Per-CP-Cy5.5 695/40 690LP
FITC, BB515 530/30 505LP
SSC 488/10  
FSC    
Yellow/Green (561 nm) PE-Cy7 780/60 750LP
PE-Cy5 670/30 650LP
PE-CF594 610/20 600LP
PE 586/15 570LP
Red (637 nm) APC-CY7 780/60 750LP
AF-700 710/50 690LP
APC 670/30 650LP

Manufacturer Site

BD FACSAria Fusion Cell Sorter

Two BD FACSAria Fusions cell sorters are configured to sort up to 20 parameters (18 fluorescence and 2 scatter). A choice of nozzles allows users sort a wide range of particle sizes. Nozzles are available in four sizes: 70, 85, 100, and 130 microns. Bulk, single cell and index sorting are available.

Practical and theoretical training is available for the Fusion, as well as experimental and data analysis support.

Configuration Collection Vessels

TUBES PLATES SLIDES
5 ml Falcon 6 well Standard
15 ml Falcon 12 well Frosted End
Eppendorf 24 well  
48 well  
96 well  
384 well  
Terasaki  
LASER CHANNEL BANDPASS FILTER LONGPASS MIRROR
UV (355 nm) BUV737 730/45 690LP
BUV496 515/30 450LP
BUV396 379/28  
Violet (402 nm) BV786 780/60 750LP
BV711 670/30 690LP
BV650 670/30 635LP
BV605 610/20 600LP
BV480 470/14 450LP
BV421 431/28 410LP
Blue (488 nm) Per-CP-Cy5.5 710/50 690LP
FITC 530/30 502LP
SSC 488/10  
FSC    
Yellow/Green (561 nm) PE-Cy7 780/60 750LP
PE-Cy5 670/30 650LP
PE-CF594 610/20 600LP
PE 586/15 570LP
Red (637 nm) APC-CY7 780/60 750LP
AF-700 710/50 690LP
APC 670/30 650LP

Manufacturer Site

BD FACS Aria II Cell Sorter

The BD FACSAria II cell sorter is configured to sort up to 17 parameters (15 fluorescence and 2 scatter). A choice of nozzles lets users sort a wide range of particle sizes. Nozzles are available in four sizes: 70, 85, 100, and 130 microns. Bulk, single cell and index sorting are available.

Practical and theoretical training is available for the Aria II, as well as experimental and data analysis support.

Configuration Collection Vessels

TUBES PLATES SLIDES
5 ml Falcon 6 well Standard
15 ml Falcon 12 well Frosted End
Eppendorf 24 well  
48 well  
96 well  
384 well  
Terasaki  
LASER CHANNEL BANDPASS FILTER LONGPASS MIRROR
UV (355 nm) BUV737 740/45 690LP
BUV395 379/28  
Violet (405 nm) BV650 655/20 630LP
BV605 615/30 555LP
AmCyan 515/20 505LP
BV421 450/50  
Blue (488 nm) Per-CP-Cy5.5 695/40 600LP
FITC, BB515 525/50 500LP
SSC 488/10  
FSC    
Yellow/Green (561 nm) PE-Cy7 780/60 750LP
PE-Cy5.5 695/40 635LP
PE-CF594 615/30 600LP
PE 586/12  
Red (637 nm) APC-CY7 780/60 750LP
AF-700 730/45 690LP
APC 670/14  

Manufacturer Site

Attune NxT Flow Cytometer

The Attune NxT Flow Cytometer is a compact, benchtop cell analyzer that has been configured with four lasers to run and analyze multicolor panels of up to 16 colors (14 fluorescence and 2 scatter). An integrated autosampler provides rapid, fully automated sample acquisition from 96- and 384-well microtiter plates.

Practical and theoretical training is available for the Attune, as well as experimental and data analysis support.

LSER CHANNEL BANDPASS FILTER LONGPASS FILTER
Violet (405 nm) VL1 450/40 417LP
VL2 525/50 495LP
VL3 610/20 555LP
VL4 660/20 635LP
VL5 710/50 680LP
VL6 780/60 740LP
Blue (488 nm) BL1 530/30 495LP
BL2 695/40 555LP
SSC    
Yellow/Green (561 nm) YL1 585/16 577LP
YL2 620/15 600LP
YL3 780/60 650LP
Red (637 nm) RL1 670/14 654LP
RL2 720/30 690LP
RL3 780/60 4740LP

Manufacturer Site

 

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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 500 nm and 20 μm can be run. Samples can be provided in 5-ml Falcon tubes, 15-ml 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 (25 mM), EDTA (1 mM) or DNAse (10 U/ml) to avoid clumping. If you are concerned that your sample is too concentrated, bring in some media to dilute your sample. 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.