Oxford Chipcytometry Days

OxfordHome

The Chipcytometry User Meeting, Wednesday, December 2nd 2015

University of Oxford, Peter Medawar Building for Pathogen Research

Chipcytometry is a fairly new technology enabling deep immune cell phenotyping and biobanking of precious (clinical) samples. Chris Willberg’s group participates in Zellkraftwerk’s Early Access program and the team has gained first-hand experience using Chipcytometry since early 2015 for a range of projects. In a user-meeting kind of workshop first-hand information among researchers was shared.

[CLICK ON THE TITLES TO DOWNLOAD THE TALKS]

Later-on, attendees had the chance to have a look at the Zellscanner ONE at Chris Willbergs Lab:

OxfordScanner

Mission accomplished: Longterm cellculture on ZellSafe chips works :-)

We often have been asked if it would be possible to seed cells on our chips and let them grow until confluency and perform assays followed by Chipcytometry on these cells. Well, in theory it should work since the chips are equipped with standard microfluidic connectors. So we gave it a try: We seeded 16 HDE cells on the chips, connected a microfluidic pump to the chip and allowed the medium to recirculate between chip and reservoir for 72h in an incubator.

This image shows the cells after seeding:

image

 

 

 

 

 

After 72h, the cells nicely formed an confluent cell layer:

livecell

 

 

 

 

 

So let’s use it for some fancy stuff…

What sample sources are best for ZellSafe chips?

One of the most often asked questions in regard to cellular samples and Chipcytometry is about the best way to prepare cells from sample source or which sample sources can be analyzed using Chipcytometry.

In general, for the analysis of cellular biomarkers our aim should be to have a 100% correlation of the amount of the in-vitro messured cellbound biomarkers compared to the in-vivo situation. This aim is not achievable, since all preparation methods like Ficoll density centrifugation, enzymatic digestion of tissues, DMSO-based freezing of cells or erythrozyte lysis significantly change the

– number or conformation of biomarkers on the cell (e.g., Ficoll reduces chemokine rezeptors)

– the composition of cells within the sample (e.g., T-cells survive freezing much better then neutrophils)

– number of cells available for analysis (e.g., centrifugation can result in a loss of 90% of cells in samples with very low cell numbers)

So here is our ranking for the best sample sources for cellsolutions and tissues:

A) Cellsolutions
1. Live cells. Best correlation between in-vivo and in-vitro biomarker expression. However, cells are changing in-vitro, so there is a short time-frame for reliable analysis. For analyzing live cells (also for calcium flux or migration assays) , a Zellscanner (available from September 2013) at your facility is a prerequisite

2. Formaldehyde (PFA)-fixed cells: Best way to produce store-able, comparable, and ship-able samples. We recommend this sample source when using our analysis services or for long-term storage. Some antibodies will not work any more after PFA-fixation due to change in epitope conformation, but alternative clones are often available that overcome this hurdle.

3. Frozen cells: Significant change of biomarker expression and cell composition. Not recommended.

B) Tissues

1. Cryosections. Best way to preserve biomarkers in tissues. We very much recommend cryosections as sample source of choice in Chipcytometry on tissues.

2. Paraffin sections: Paraffin embedding and demasking by heating is very much changing biomarker expression, also leads to a high unspecific background when staining with Fluorescence-labelled antibodies. Not recommended.

3. Digest tissue: We have recently published how much tissue digestion can change biomarker expression and function of cells leading to biased data. Not recommended.

ZELLKRAFTWERK Announces Early Access Program

Select Drug Developers to obtain pre-release access to groundbreaking cellular biomarker discovery platform.

Hannover / Germany, January 3, 2013 – ZELLKRAFTWERK today announced the start of its early technology access program for Chipcytometry, a groundbreaking high-content cytometry technology for cellular biomarker discovery and bioanalysis. Chipcytometry allows for sample storage and biomarker preservation for about 12 month and enables iterative re-analysis of the same sample with different marker-sets for several times. In a constantly growing number of studies, Chipcytometry has been validated for a broad range of clinical specimens and has delivered excellent results published in high rank journals.

Pharmaceutical researchers in many disease areas currently have high interest in revealing correlations between biomarker expression and signaling pathway activity directly on cells from tissues and body fluids and the respective patient outcome. Chipcytometry is an extremely powerful tool for cellular biomarker discovery in the field of drug development and uses proprietary technology to make life science research and drug and diagnostic development programs safer, more efficient and effective. Whether measuring dozens of biomarkers or simply a few, our approach provides reproducible, quantitative, sensitive immunophenotyping data from a small sample volume.

ZELLKRAFTWERKs early technology access program now makes Chipcytometry available to researchers and drug developers worldwide. Under this program ZELLKRAFTWERK will work with academic institutions and biotech companies in specific disease areas on an exclusive basis to identify new drug target and new efficacy and safety biomarkers.

Since 2008, Dr. Christian Hennig and his team has used Chipcytometry in a growing number of preclinical and clinical applications and has published excellent results in high rank peer-reviewed journals. ‘The massive multiplexing capability really sets Chipcytometry apart from conventional flow cytometry‘ states Christian Hennig, CEO and co-founder of ZELLKRAFTWERK. ‘For the first time Chipcytometry allows researchers to use an iterative ‘stain-think-stain’-approach for biomarker discovery. It’s really like you are communicating with your cells‘ he adds.

About Chipcytometry:

The Chipcytometry technology platform provides hardware and software for explorative cytometry. It differentiates itself from flowcytometry by keeping cells immobilized inside a microfluidic-channel allowing the stepwise, quantitative analysis of biomarkers on cells and tissues using a flexible and literally unlimited set of markers. Chipcytometry has been developed by a team of researchers at Hannover Medical School. The project is funded by Federal Ministry of Education and Research (Germany) under the GOBio program.

The platform is compatible to a broad range of biomarker types (baseline, activity, and functionality biomarkers) and allows for multiplexing of up to 30 biomarkers per sample in an iterative workflow. Chipcytometry has shown a 10-fold better sensitivity compared to flowcytometry. Sample- and cellular biomarker stability has been validated for 12 month when cells are stored inside their proprietary microfluidic chips. The Chipcytometry platform recently has been validated for clinical use in a number of biomarker discovery projects with institutions like MD Anderson Cancer Center, Houston, German Competence Network Multiple Sclerosis, the German Center for Lung Research and the Radboud University, Nijmegen.

Press contact:

Gary Jones, Business Development; Fon: +1 (617) 500-8221

Web: www.zellkraftwerk.com Email: jones@zellkraftwerk.com