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Northwestern University Feinberg School of Medicine
Skin Biology & Diseases Resource-Based Center
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Translational & Experimental Skin Testing & Immune Tracing Core

The Translational & Experimental Skin Testing & Immune Tracing (TEST IT) Core is directed by Caroline Le Poole, PhD, and co-directed by Kurt Q. Lu, MD. TEST IT Core focuses on the important relationship between keratinocytes and the skin immune system and provides instrumentation and immunological assay services and the unique opportunity for human testing in a controlled environment.

Services

Ordering is done through NU Core and cost information is available via our core price sheet (PDF).

  • Provide service, equipment, protocols and training to help investigators differentiate healthy skin from skin with abnormal immune responses at the cellular and molecular level, to enable biomarker identification as well as to increasing our knowledge of skin immune responses
  • Homogenization and cell derivation from fresh tissues while preserving markers on the cell surface
  • Characterization of cell isolates at the protein levels by cytokine arrays, ELIPOTS and multi-parameter cell scanning or sorting
  • DNA or RNA analysis of captured tissue or single cells, including paired TCR subunit sequences  
  • Purification of immune cell populations to be combined with extracellular matrix composites containing keratinocytes, melanocytes and fibroblasts (genetically modified or not) in a 3D setting
  • Support for state-of-the-art multispectral analysis and imaging of diseased skin and skin reconstructs

Equipment

 Leica BondRx Automated Stainer

Functions

  • Fully automated IHC and ISH Research Staining.
  • In a single FFPE tissue section and frozen samples.
  • Monoplex and Multiplex Staining.
  • Opal kits measure up to 6 tissue biomarkers simultaneously plus a nuclear stain.
  • Retain spatial cellular context that is lost when using other methods.
  • Opal reagents utilize tyramide signal amplification (TSA) to increase antigen marker intensity by 10-100-fold compared to conventional indirect immunofluorescence methods.
  • Opal fluorescent detection substrates bind covalently near the epitope, allowing subsequent removal of antibodies to clear the tissue for detection of the next target. The signal remains intact after antibody removal.
  • A manual 4-color Opal staining protocol typically takes one day to complete and an automated 7-color can be run overnight, with all targets being imaged simultaneously.

Features

  • Accelerate test programs through speed, efficiency, and consistent automation, reducing manual work.
  • Add signal amplification to almost any immunoassay. Improve sensitivity by 10 to 100-fold. 2 to 4 logs of Amplification dynamic range versus 1 log for chromogenic. And signal is more linear.
  • Excellent resolution with low background.
  • Reduce antibody consumption.
  • Unmodified primary antibodies
  • Use the best primary antibodies, regardless of species.
  • Multispectral imaging eliminates fluorophore crosstalk and interference from tissue autofluorescence, allowing precise measurement of each fluorescence signal within a tissue sample.
  • High-plexing for multiple biomarker detection strategies. The stain quality and reproducibility are better than the competition.

Services Provided

  • Consultation with your research projects.
  • Optimizations on the protocols.
  • Monoplex-multiplex staining up to 6 tissue biomarkers simultaneously plus a nuclear stain.
  • One batch of automated staining simultaneously up to 30 pieces of tissue slides.

Notes

  • Staff-assisted service only

 Vectra Multispectral Imager

Functions

  • Vectra imaging systems use multispectral unmixing to provide quantitative results for each marker in a multicolor IHC slide, making them ideal for imaging and analysis.
  • Visualize and quantify the complex interactions between immune and tumor cells across whole tissue sections with whole-slide and multispectral imaging.
  • When combined with inForm analysis software, this novel solution enables automated cellular phenotyping and tissue segmentation for virtually unlimited investigation of the biology expressed in your samples.
  • The complete workflow – from immunostaining, through imaging and analysis – is known as Phenoptics.

Features

The Vectra 3 automated quantitative pathology imaging system, together with the inForm software:

  • Provides multispectral and automated multi-slide imaging. Visualizes, analyzes, quantifies and phenotypes immune cells in situ in solid tissues
  • Detects and measures multiple weakly expressed and overlapping biomarkers within a single H&E, IHC or IF tissue section and in TMAs
  • Automated loading of up to 6 slides
  • Phenochart whole slide viewer for slide navigation provides context for identification of regions of interest within a digital pathology workflow
  • Automatic identification of specific tissue types using integrated inForm analysis software
  • Quantitative multi-spectral imaging, seamless image tiling, and minimal photobleaching Provides multispectral and automated multi-slide imaging. Visualizes, analyzes, quantifies and phenotypes immune cells in situ in solid tissues

Services Provided

  • Multiplex (Vectra) imaging acquisition
  • Quantitative measurement of multiplex imaging
  • Instrument and Software training available for clients

Notes

  • Staff-assisted service
  • Self-service once the user is trained by staff

 inForm: Multiples image analysis software

Functions

inForm is a patented automated image analysis software package for accurately visualizing and quantifying biomarkers in tissue sections. Two standalone versions of inForm image analysis software are available inForm Cell Analysis and inForm Tissue Finder.

  • inForm Cell Analysis enables per-cell analysis of immunohistochemistry (IHC) and immunofluorescence samples, allowing the separation and measurement of weak and spectrally overlapping markers in single and multiplex assays.
  • inForm Tissue Finder automates the detection and segmentation of specific tissue types using patented user-trainable algorithms developed to recognize morphological patterns.
  • Data may be exported from inForm for additional analysis such as distance mapping between selected cellular phenotypes.
  • In addition to fluorescent composite images, inForm provides simulated brightfield monoplex IHC from the same data for improved visual interpretation.

Features

  • Visualizes, analyzes, quantifies and phenotypes immune and other cells in situ in solid tissue.
  • Pathology views renders immunofluorescence images as H&E, DAB and hematoxylin.
  • Separates weakly expressing and overlapping markers.
  • Cellular analysis of H&E, IHC and immunofluorescence in FFPE tissue sections.
  • Automatic identification of specific cell and tissue compartment types using trainable feature-recognition algorithms.

Services Provided

  • Image analysis: Quantify Biomarkers in Tissue Sections.
  • Provide Inform Software training for clients.

Notes

  • Staff-assisted service
  • Self-service once the user is trained by staff

 IsoLight: Single Cell proteomics

Functions

The IsoLight is a hub for comprehensive functional profiling of each cell type across a large assay menu of single-cell chip and software products. Run an entire highly-multiplexed proteomics workflow. achieving fully analyzed data of single cell’s highly multiplexed secretome or intracular proteome on the same day.

  • Single cell proteomics uniquely finds correlates that lead to biomarker breakthroughs. Uncover the functional subsets of cells and unlock powerful correlative data.
  • Single-Cell System uniquely reveals the most potent functional cell subsets that secrete multiple cytokines simultaneously at high intensity

Features

Automate test Single Cellular Proteomics with high quality results, while saving your time and resources:

  • Single-cell functional phenotyping for innate & myeloid cell types.
  • Reveal the functional sources of inflammation from rare polyfunctional subsets of cells in an automated manner.
  • Measure cellular protein to protein interactions and adaptive resistance pathways.
  • Harnessing the power of functional proteomics.

Services Provided

Single-cell Secreted Protein Assay and Analysis: 

  • Polyfunctional Strength
  • Innate & Myeloid
  • Polyfunctional Inflammation
  • Tumor Signaling
  • Functional phenotyping of immune cells
  • Proliferation/ Cytotoxicity assay.

Up to 30 cytokines, 8 isocode chips with a throughput up to 8K cells per run can be loaded.

Notes

  • Staff-assisted service
  • Self-service once the user is trained by staff

 IsoSpeak software: Single Cell Proteomic analysis

Function

  • Automatically translate biomarkers and utilize intuitive visualizations to understand correlations

Feature

  • Data visualization: Automated on-site analysis and advanced, functional, single cell mapping with IsoSpeak software

Services Provided

  • IsoLight results analysis
  • Provide isoSpeak Software training for clients.

Notes

  • Staff-assisted service
  • Self-service once the user is trained by staff

 MSD: Meso Quickplex SQ 120

Functions 

  • Meso Scale Discovery - MSD is a method similar to ELISA except MSD uses electrochemiluminescence (ECL) as a detection technique as opposed to a colorimetric reaction employed by ELISA. 
  • An ECL system platform has many advantages over a traditional ELISA system.

Features

Key advantages over a traditional ELISA system:

  • Higher Sensitivity.
  • Better Dynamic Range.
  • Less Matrix Effects.
  • Ability to Multiplex panels.
  • Ability to Scale Up.
  • Better Efficiencies.
  • Less sample volume requirement.
  • Rapid throughput

Services Provided

  • Cytokine test and analysis
  • Monoplex-multiplex panels.
  • One well sample tested simultaneously up to10 biomarkers.

Notes

  • Staff-assisted service
  • Self-service once the user is trained by staff

 CyTEK: High-dimensional spectral immune-profiling

Functions 

  • Cytek Aurora is a prodigy incorporating a unique combination of patent-pending innovative technologies that takes flow cytometry to the next level of performance and flexibility.
  •  Newly build spectral analyzer with a unique capability of detection over the full emission spectra.
  • With up to three lasers, three scattering channels, and 64 fluorescence channels, the Aurora suits every laboratory’s needs, from simple to high complexity applications.
  • A paradigm shifting optical design provides unprecedented flexibility, enabling the use of a wide array of new fluorochrome combinations without reconfiguring your system for each application.
  • The state-of-the-art optics and low-noise electronics provide excellent sensitivity and resolution. Flat-top beam profiles, combined with a uniquely designed fluidics system, translate to outstanding performance at high sample flow rates.

Features

The end result is a system that delivers high quality data where rare and dim populations are easily resolved regardless of assay complexity

  • Maximum Channels: 64 fluorescence channels.
  • Maximum Colors: Beyond 40 colors demonstrated including fluorochromes with emission spectra in close proximity to each other.
  • Maximum Sensitivity: Sensitivity redefined using state-of-the-art optics and low-noise electronics.
  • Maximum Flexibility: No changing optical filters for any fluorochrome. Use any commercially available fluorochrome excited by the onboard lasers.
  • Maximum Acessibility: A powerful, high value system that is accessible to a wide range of users.

Services Provided

  • Comprehensive tests with high multiplexing.
  • Multiparameter FACS staining and analysis.
  • Instrument and Software training available for clients.

Notes

  • Staff-assisted service 

 10X Genomics Chromium System: Single cell genomics

Functions 

  • By traditional bulk RNA-seq methods, cell-to-cell differences are missed and cellular heterogeneity can be completely masked
  • Single cell RNA-Seq opens a whole new field in biology by allowing the study of cell-to-cell transcriptome heterogeneity.
  • Examines thousands of cells per sample through cell-by-cell 3’ end counting of mRNA transcripts.
  • Enables identification of rare cell types, analysis of cellular heterogeneity, and understanding of individual cell states and cellular responses to environmental signals and conditions.

Features

  • Analyze transcriptomes on a cell-by-cell with Chromium Single Cell 3’ Solution
  • Samples: Single cell suspension; total RNA; tissue (for nuclei isolation).
  • To ensure high-quality results Dead Cell Removal has optimized
  • High-Throughput 10x Genomics Chromium Single Cell RNA-Seq: process 100’s to 10,000’s of single cells in a single 7-minute Chromium Instrument run.
  • To ensure high-quality results Dead Cell Removal has optimized

Services Provided

  • Single cell RNA sequencing and data visualization analysis
  • Consultation with single cell preparation.
  • Generate libraries for (a) 5’ gene expression analysis, (b) T cell receptor repertoire profiling, and (c) B cell immunoglobulin repertoire profiling

Notes 

  • Staff-assisted service
  • Self-service once the user is trained by staff

 GentleMACS Octo Dissociator: Cell Separation

Functions 

With tissue-specific enzyme kits:

  • Validated, optimized dissociation of human or mouse tissue.
  • High yields - Preserved membrane integrity and surface epitopes.
  • Fast. Optimized protocols. Gentle dissociation. Efficient. high standardization. Reliable

Features

  • Semi-automated and standardized tissue dissociation or homogenization of up to eight samples.
  • Single-use the unique gentleMACS Tubes allow sample preparation in a closed and sterile system. 
  • In addition to the ready-to-use gentleMACS Programs, the gentleMACS Octo Dissociator allows the user to develop individualized programs for almost any biological material.

Services Provided

  • Dissociation of tissues into single-cell suspensions, including:
    • mouse or human tumor; mouse or rat neonatal heart; neural tissue; mouse spleen, lung, lamina propria, muscle, epidermis, or liver; mouse or human skin; and more.
    • total RNA or mRNA isolation from fresh or frozen samples; protein extraction; determination of bacterial or viral load; mitochondria isolation

Notes

  • Staff-assisted service
  • Self-service once the user is trained by staff

 Controlled Rate Freezer: Ultra Low Freezer

Functions 

  •  The controlled rate freezers are sample preparation devices used for the preparation of samples for storage at cryogenic temperatures (-130°C or below).
  • The Controlled-Rate Freezer can cryopreserve your cells and maintain high cellular viability via controlled-rate freezing.

Features

  • The freezing system delivers liquid nitrogen into a closed chamber into which the cell suspension is placed.
  • Careful monitoring of the rate of freezing helps to prevent rapid cellular dehydration and ice-crystal formation

Service Provided

  • Sample storage with temperature precisely controlled to get the cells down to the cryopreservation temperature without damage: Temperature Range‎ ‎50°C to -180°C

Notes 

  • Staff-assisted service
  • Self-service once the user is trained by staff

Core Leadership

Caroline Le Poole, PhD
Core Director
Professor of Dermatology and Microbiology-Immunology

Kurt Q. Lu, MD
Associate Core Director
Eugene and Gloria Bauer Professor of Dermatology

Stephanie Rangel, PhD
Director of Clinical Trial Unit, Dermatology
Research Assistant Professor of Dermatology

Contact Us

We welcome your interest in our core. Please contact Madeleine Belback, research study coordinator, for more information.

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