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Cell Profiling

National facility

The Cell Profiling facility enables researchers to access the resources and expertise from the Human Protein Atlas and the primary aim is to do full service immunofluorescence projects at moderate to high-throughput level. Based on the resource of more than 25,000 antibodies and a panel of >20 different human cell lines, this facility has a unique position to investigate subcellular spatial proteomics for human and rodent biology. The facility is also currently implementing highly multiplex imaging (>15 markers) of tissue sections using the CODEX platform: https://www.scilifelab.se/facilities/cell-profiling/codex

What if you had access to:

  1. 25,000 validated antibodies towards human targets
  2. A good confocal and screening microscope
  3. A broad panel cell lines of different origin
  4. Dedicated people with great experience of generating immunofluorescence data

As you would only pay for the amount of antibody you use, the price per antibody is about 10 % of the vendor price, which allows you to stain for many more protein targets at a reasonable price.

What would you do?

Contact us at cellpro.facility@scilifelab.se to discuss how can your project benefit from this unique resource!

SERVICES

We perform everything from project design, sample preparation and imaging to give you as good results as possible. We share infrastructure and resources with the Human Protein Atlas and this include a proteome wide library of antibodies that can be used in tailor made projects to answer specific biological questions. You are also welcome to use our microscopes for imaging after an introduction with one of the staff scientists.

Proposed projects are assessed and prioritized according to the following model

Initial screen of project by facility:

1. Technical feasibility (yes/no)
2. Capacity requested for the project (classification: small/medium/large)

Project prioritization by committee based on:

1. Scientific potential 0 - 5
2. Supporting preliminary data 0 - 5
3. Supports facility development (competence and techniques) 0 - 5
4. Significance of facility specific technique for project 0 – 2

APPLICATIONS

  • High-throughput profiling of protein targets in single cells using multiplexed immunofluorescence
  • Comparison of protein expression and localization in different cell states
  • Phenotypic screens
  • Immune cell distribution in tissue samples
  • Antibody validation and testing for further screens

Here are a few examples of previous projects:

  1. Explore the effect of a small molecule/substance on a particular organelle, family of proteins or proteins within a defined pathway.
  2. Screen for proteins co-localizing with a defined marker in different cellular conditions like autophagy, serum starvation or hypoxia.
  3. Confirm data from mass spectrometry experiments, such as translocation of proteins
  4. Evaluation of immune cell infiltration in muscle biopsies from diabetic patients before and after physical exercise
  5. Validate knock-out cell lines and antibodies by evaluation of fluorescence signal reduction and quantitative image analysis

 

EQUIPMENT

  • Antibodies. 25,000 ICC validated antibodies from the Human Protein Atlas project.
  • Confocal microscope. 3 LEICA SP5 up-right confocal microscopes, 1 inverted SP8 confocal microscope, 1 DMI8 inverted screening fluorescence microscope
  • Liquid handling. Two TECAN FREEDOM EVO 150 pipetting robots. One TECAN Fluent 480.
  • CODEX platform, enabling highly multiplex imaging (>15) at single cell level in tissue sections

RECENT PROJECTS

  • A screen of >100 nucleolar proteins in a model for ALS (amylotropic lateral sclerosis) to evaluate general morphology and function of nucleoli in normal and diseased ALS state. The hundred nucleolar protein targets were also used to confirm the effect of a small molecule identified in a previous screen, against the accumulation of toxic peptides in the nucleoli. Cell Chemical Biology (doi:10.1016/j.chembiol.2018.10.020).
  • A four-stage isogenic human cell model for malignant transformation was characterized with RNA-seq to identify differentially expressed genes. Subsequent functional analysis of the target proteins in the cell model was done at a single cell level using antibodies and confocal microscopy. We could in this manner characterize the molecular mechanism behind malignant transformation in this model and validate potential novel biomarkers of clinical interest. Danielsson et al, PNAS (Apr 2013).
  • A panel of 27 monoclonal antibodies were validated in different cell lines to evaluate their functionality as organelle markers for immunofluorescence applications. The antibodies validated by the Cell profiling Facility are now part of the Atlas Antibodies product portfolio.  https://atlasantibodies.com/#!/products/panel/organelle-markers