Zellscanner One 칩사이토메트리(Chipcytometry)
ONE PLATFORM – MANY APPLICATIONS
At ZELLKRAFTWERK, we offer complete workflows for high-content cytometry on cells and tissues, not just stand-alone solutions like reagents, robots, or software.
The basic principles of the fourth industrial revolution are applied to provide products and services for the automated analysis of fluorescent biomarkers on living cells or long-term stored biobanked cells and tissues.
We also offer a wide range of products and services that cover the complete workflow, from sample preparation and biobanking to biomarker analysis and data mining.
HIGH QUALITY SAMPLES = HIGH QUALITY BIOMARKER DATA
ChipCytometry enables the analysis of a virtually unlimited number of biomarkers on a single- tissue or cell suspension sample [Analysis].High quality sample sources such as blood or fresh frozen tissues and gentle sample preparation procedures result in better biomarker data quality. We recommend that samples be ideally prepared on-site. After preservation, samples can be shipped under conditions that prevent biomarker degradation during storage.
At ZELLKRAFTWERK, we have developed reagents, methods, and hardware that provide our customers with a revolutionary new way of the controlled, long-term biobanking of cells and tissue sections with exceptional biomarker preservation through ZellSafe chips. Samples can be easily prepared and preserved on ZellSafe chips even in remote locations and stored/shipped for controlled centralized analysis without significant biomarker degradation.
LONG-TERM SAMPLE PRESERVATION ON ZELLSAFE BIOREPOSITORIES
ZellSafe chips are small biorepositories with standardized connectors that can be handled by technicians or study nurses as well as by pipetting robots. Cell suspensions and tissue sections are prepared and deposited in the biorepositories according to standard operating procedures [SOP].
The samples are immobilized on the surface of the chip. For long-term storage, fixatives are added to prevent biomarker degradation. We offer a growing range of ZellSafe chips to meet the needs of different types of cells and tissues:
WE CALL IT TrueBiobanking
A look at the biomarker preservation data of the ZellSafe biorepositories shows that the cells and tissues remain intact and there is almost no cell loss after at least 24 months of storage. The impact of the storage effects of biomarker loss during this period was a maximum 5% compared to day 0. We are the first to achieve such low values, therefore giving rise to the name TrueBiobanking.
Loss of material or biomarkers after 2 years of storage
At ZELLKRAFTWERK, we have developed an advanced form of iterative imaging cytometry called ChipCyometry that is used for analyzing cells and tissues inside ZellSafe longterm biorepositories [Sample].
• Advanced imaging: HDR imaging, 3D cell recognition, specially designed filter sets for high-efficiency, protein-protecting fluorescence
detection with unprecedented sensitivity
• Non-destructive analysis: The sample is not consumed during analysis
• Recovery of detection channels: unlimited number of biomarkers per sample
• Mixed parallel and serial marker detection: no or very low panel set-up time compared to high-plex flow panels
For samples to be analysed within a short period of time with a maximum number of biomarkers, the most rapid means is theoretically by parallel analysis. However, simultaneous analyses of a large number of biomarkers result in problems associated with antibody-antibody, antibody-dye, and dye-dye (e.g., agglomerate formation or spillover) interactions in the cocktail mix. This may lead to an extensive set-up time for new cocktail mixtures or even impossible to combine marker combinations. Yet, individual serial analysis of single markers does not require a set-up time but is much slower. As a compromise, we have introduced mixed serial and parallel measurements:
ZELLKRAFTWERK develops and validates max 5-plex assays. The processing time is usually no longer than 1 week. Each of these 5-plex assays can be combined with one another and individually serially stained.
The set-up of multicolor cocktails is described in further detail below:
As a result of the sample storage capabilities (no ad-hoc analysis necessary) and assay development, the ChipCytometry workflow differs to that of conventional flow cytometry:
1. The chronological separation of sample acquisition and the time point of analysis enable the valid, controlled sending of
samplesand centralized analysis, even with global sample acquisition.
2. Samples can be tested in a screening phase with a virtually unlimited number of markers using the serial single-color
approachthat requires a very low set-up time.
3-4. Throughput can be later increased using serial analysis of 5-plex assays and/or switch antibodies that do not require bleaching
of remaining fluorescence [available on request].
Generated data is useless without knowledge about how the data was generated, what the data points mean, and how the data was transformed after initial acquisition. Reuse, sharing, and comparison of datasets are impossible without access to accompanying metadata. Therefore, ZELLKRAFTWERK has integrated complete metadata recording for ALL processes from sample preparation through storage and analysis to data processing. Based on the principles of the semantic web and ontologies, we developed a description language (EDL – experiment description language) that records every object and process involved in ChipCytometry. This is useful in a number of situations:
1. Electronic lab book: All metadata (e.g., time points, durations, temperatures) are collected either automatically or via standardized
user interfaces, leading to the automatic generation of an electronic lab book.
2. Seamless integration of all machines that understand and respond to EDL: All machines integrated into our ChipCytometry
products understand and respond to EDL. Therefore, new, more complex workflows can be realized by the software – applying
therules of Fourth industrial revolution.
3. Plug-and-play data processing with integrated, EDL-driven interface to R statistic computing language
4. Easy forwarding of all necessary metadata to downstream data analysis pipelines (flow cytometry software such as FlowJo®,
pathology software such as Definiens Tissue Studio®
Publications Describing Aspects of the Technology
A versatile platform for comprehensive chip-based explorative cytometry
Hennig C, Adams N, Hansen G. Cytometry A. 2009.READ MORE
Publications Using Chipcytometry
TCR and Inflammatory Signals Tune Human MAIT Cells to Exert Specific Tissue Repair and Effector Functions
In this Cell paper, the authors show that activation of human MAIT cells is TCR-dependent or TCR-independent, enhanced by TL1A and that the TCR-dependent triggering induces a tissue repair program.
Long-term in vivo microscopy of CAR T cell dynamics during eradication of CNS lymphoma in mice
In this PNAS paper, the successful attack of a CNS lymphoma by CAR T cells was demonstrated in a mouse model.READ MORE
A preliminary study for the assessment of PD-L1 and PD-L2 on circulating tumor cells by microfluidic-based ChipCytometry
In this study, David Skibinski and colleagues from Merck Singapore developed a method that applies ChipCytometry for the detection and phenotyping of circulating tumor cells in blood.READ MORE
Gain-of-function STAT1 mutations are associated with intracranial aneurysms
Dadak M, Jacobs R, Skuljec J, Jirmo AC, Yildiz Ö, Donnerstag F, Baerlecken NT, Schmidt RE, Lanfermann H, Skripuletz T, Schwenkenbecher P, Kleinschnitz C, Tumani H, Stangel M, Pul R. Clin Immunol. 2017.READ MORE
Monocyte/macrophage lineage commitment and distribution are affected by the lack of regulatory T cells in scurfy mice
Skuljec J, Cabanski M, Surdziel E, Lachmann N, Brennig S, Pul R, Jirmo AC, Habener A, Visic J, Daluege K, Hennig C, Moritz T, Happle C, Hansen G. Eur J Immunol. 2016.READ MORE
a-NAC-specific autoreactive CD8+ T cells in atopic dermatitis are of an effector-memory 2 type and secrete IL-4 and IFN-y
Antigen-specific T cells were detected and phenotyped using the rare cell detection feature of ChipCytometry and MHC class I specific tetramers on living cells immobilized inside XXL chips.
Roesner LM, Heratizadeh A, Wieschowski S, Mittermann I, Valenta R, Eiz-Vesper B, Hennig C, Hansen G, Falk CS, Werfel T. Journal of Immunology. 2016.READ MORE
Der p1 and Der p2-specific T cells display a Th2, Th17, and Th2/Th17 phenotype in atopic dermatitis
Roesner LM, Heratizadeh A,Begemann G,Kienlin P,Hradetzky S,Niebuhr M,Eiz-Vesper B,Hennig C,Hansen G,Baron-Bodo V,Moingeon P,Werfel T. Journal of Investigative Dermatology. 2015.READ MORE
Pulmonary transplantation of macrophage progenitors as effective and long-lasting therapy for hereditary pulmonary alveolar proteinosis
In this excellent study, both single cell and tissue cytometry options of ChipCytometry were applied to implement a high-content in-depth analysis of transplanted cells in a humanized mouse model.
Happle C, Lachmann N, Skuljec J, Wetzke M, Ackermann M, Brennig S, Mucci A, Chari AJ, Groos S, Mirenska A, Hennig C, Rodt T, Bankstahl JP, Schwerk N, Moritz T, Hansen G. Science Translational Medicine. 2014.READ MORE
Successful treatment of autoimmune and lymphoproliferative complications of patients with intrinsic B-cell immunodeficiencies with Rituximab
Hennig C, Baumann U, Ilginus C, Horneff G, Foell J, Hansen G. Br J Haematol. 2010.READ MORE
Identification and quantification of basophils in the airways of asthmatics following segmental allergen challenge
Dijkstra D, Hennig C, Hansen G, Biller H, Krug N, Hohlfeld JM. Cytometry A. 2014.READ MORE
Allergy prevention starts before conception: maternofetal transfer of tolerance protects against the development of asthma
ChipCytometry was used to analyze the antigen-specific immune response in the offspring of mice tolerized to a specific allergen. Surprisingly, interferon-γ producing T cells were found to be induced in the offspring due to diaplacentar immune complex transfer.
Polte T, Hennig C, Hansen G.J Allergy Clin Immunol. 2008.READ MORE
CD3+CD20+ T cells in peripheral blood: Proper flow cytometry is a prerequisite of exact phenotyping
ChipCytometry was applied as a supportive technology to confirm flow cytometry results that demonstrated the existence of T cells expressing CD20.
Wilk E, Witte T, Marquardt N, Hennig C, Hansen G, Schmidt RE, Jacobs R. Arthritis Rheum. 2010.READ MORE
Lung function and inflammation during murine Pseudomonas aeruginosa airway infection
Woelbeling F, Munder A, Kerber-Momot T, Neumann D, Hennig C, Hansen G, Tuemmler B, Baumann U. Immunobiology. 2011.READ MORE