SAMPLE - ANALYSIS - RESULT

SAMPLE

HIGH QUALITY SAMPLES = HIGH QUALITY BIOMARKER DATA

ChipCytometry enables the analysis of a virtually unlimited number of 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:

zellsafechips_5
zellsafechips_5_text

Load cells/tissue onto the chip
Each cell has a permanent individual address

Add fixation buffer & storage buffer
Stops biomarker degradation

Seal chip for biomarker preservation
Long-term storage > 20 months

zellsafechips_biobanking

WATCH THE SIMPLE LOADING PROCEDURES OF THE ZELLSAFE BIOREPOSITORIES

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

DNA at -80°C
20%
Cells frozen at -80°C
80%
Cells on ZellSafe Chips at 4°C
5%

Source: Zellkraftwerk, Christian Hennig

ANALYSIS

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
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ASSAY DEVELOPMENT

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. 

analysis_assaydevelopment

The set-up of multicolor cocktails is described in further detail below:

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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 samples and 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 approach that 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].

analysis_modes

RESULT

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 the rules 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®

ANALYZING A 37-PLEX DATA SET

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