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 CHANGE IN VISITATION GUIDELINES: Please read before visiting Rutgers Cancer Institute.
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Immune Monitoring Sample Preparation

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GUIDELINES FOR SAMPLE SUBMISSION

For clinical trial samples, please contact Ankit Saxena, PhD, Managing Director (as4075@cinj.rutgers.edu) for consultation on how samples should be collected and submitted for processing and analysis. For other samples, please see below or contact us for specific guidelines to meet your protocol and analysis needs.


SUBMISSION OF SAMPLES FOR FLOW CYTOMETRY

Single-cell samples (cultured or processed tissue) should be submitted on ice in RPMI or DMEM media supplemented with 2% FBS.  One sample for flow cytometry is up to 1x107 cells in 1 ml of media.

Tissue samples that the Shared Resource will dissociate for flow cytometry should be submitted on ice in RPMI or DMEM media supplemented with 2% FBS.  Please bring in 6-well or 12-well plates, with the sample annotations and individual tissue weights provided separately.  A single sample is considered up to 1gram of tissue.

For basic research experiments on mouse samples, please provide an additional spleen sample in media for single color control preparation.

 

SUBMISSION OF SAMPLES FOR SERUM CYTOKINE ANALYSIS

Serum samples should be submitted either frozen on dry ice or thawed on wet ice.  For each sample, please provide 100 microliters of serum for analysis.  The 96-well plate format allows for 80 samples wells and 16 control / standard curve wells.  It is recommended that the samples are run in duplicate, for a total of 40 samples per plate.

If you are submitting culture media / supernatant for analysis, please provide an additional 5ml of the sterile culture media for calculation of background protein levels.

 

SUBMISSION OF SAMPLES FOR SINGLE-CELL RNA SEQUENCING

Samples for single-cell RNA sequencing should be submitted as single-cell suspensions that have been pre-filtered through a 70-micrometer filter and tested for at least 90% viability.  Cells should be suspended to a concentration of 1000-1200 cells per microliter in RPMI, or DMEM media supplemented with 2% FBS and brought to the facility on wet ice.  If additional processing is necessary to exclude dead cells or enrich for a target population, please let the facility know at the time of booking your experiment.

If whole tumor tissue (mouse or human) is being submitted for processing/dissociation following by single-cell RNA sequencing, please bring the samples on ice in RPMI or DMEM media supplemented with 2% FBS.  A single sample is considered up to 1gram of tissue.

For other tissues or preparations, please contact the Shared Resource for consultation.

 

SUBMISSION OF SAMPLES FOR NUCLEIC ACID EXTRACTION

Single-cell suspensions for nucleic acid extraction should be pelleted, flash-frozen in liquid nitrogen, then transported to the Shared Resource on dry ice. 

Fresh tissue samples should be placed in RNAlater solution according to standard protocols, and then stored at -20 until ready for analysis.  These samples should be transported to the Shared Resource on dry ice. 

Fresh tissue samples can also be flash-frozen in liquid nitrogen, and then stored at -80 degrees until ready for analysis.  These samples should be transported to the Shared Resource on dry ice.

FFPE-preserved tissue can be submitted as a tissue block.

Please contact the Shared Resource for guidelines about the submission of blood samples for DNA/RNA/cfDNA extraction.

 

SUBMISSION OF SAMPLES FOR NUCLEIC ACID ANALYSIS

DNA/RNA samples for Bioanalyzer and Qubit analysis should be transported to the Shared Resource on ice.