Cell Processing Workstation. In Situ Laser-Mediated Cell Purification and Processing

Transcription

1 LEAP Cell Processing Workstation Cell Processing Workstation In Situ Laser-Mediated Cell Purification and Processing

2 Expanding the Capabilities of Cell Processing Consistent, High-Yield, High-Purity Processing of Sensitive Cells As scientists strive to answer today s more demanding questions in cell With the LEAP workstation, stem cell colony passaging and purification, biology, the ability to work with challenging primary cells and cell lines has adherent cell purification, and cell line generation (for screening or protein become a critical need. Standard methods for purifying and processing manufacturing) can be performed with ease and confidence, making the cells flow cytometry, magnetic bead purification, etc. often cause LEAP workstation the ideal addition to any cell biology lab seeking to expand significant trauma, resulting in cell losses and dramatic changes in its cell processing and purification capabilities. morphology, physiology, and cell-to-cell interactions for the cells that do survive. These issues can complicate experimental design, execution, and interpretation, limiting research to experiments that use cells with lower Maintaining Physiologically Relevant Behavior in Cells physiological relevance. The need to develop standardized methods for purifying and processing sensitive Changes in cell morphology and function after treatment with trypsin to remove the cells from their growth surface: cells is especially important for stem cell researchers. Current methods for isolating and passaging stem cell colonies are tedious and labor intensive, and the results are highly variable due to lack of standardization. In addition, it is difficult to achieve high purity of adherent and/or colonized cell types, and cell yield and viability often suffer using current physical or enzymatic disruption methods. The new LEAP cell processing workstation overcomes these difficulties by enabling direct purification and processing of cells in situ, right where they are grown. The LEAP workstation allows cells to be identified by gating on any combination of parameters, including brightfield (morphology), multi-color fluorescence (intensity or localization), and even location (in the well or relative to other cells). Unwanted cells are eliminated by a highly accurate, rapid-pulse laser that shoots up to 1,000 cells per second. The ability to eliminate unwanted cells without disturbing them and immediately verify results allows very high purity to be obtained with very high cell yield and viability. 2 LEAP Cell Processing Workstation A B A: Neuronal cells exhibit normal axon extensions and connections to other cells under healthy growth conditions. B: After treatment with trypsin to detach the cells, there is a clear change in cell morphology, loss of connection to other cells, and decrease in cell viability. The LEAP workstation allows you to purify and process cells under physiologically relevant conditions.

3 Overview of the LEAP Workstation In Situ Cell Purification and Processing Combining high-speed, whole-well imaging and cell identification capabilities with an ultra-fast laser, the LEAP workstation provides a precise method for eliminating unwanted cells in situ, preserving cell morphology, physiology, and cell-to-cell interaction. The precision of the LEAP workstation reduces labor-intensive manual cell preparation and manipulation, reducing the variability in your cell purification and processing assays. The LEAP workstation maximizes utility with an array of capabilities: Employs ultra-fast laser optics for selective and efficient processing of cells Provides data from all cell types adherent and non-adherent Allows brightfield cell imaging, identification, and characterization Enables fluorescence imaging and identification (eight excitation and eight emission wavelengths) Analyzes every cell in every well due to elimination of brightfield edge effect Rapidly images the entire well (up to 40x faster than typical imaging systems) Requires minimal sample manipulation Operates on a variety of multiwell plate formats LEAP uses a powerful F-theta lens and galvanometer mirrors to rapidly scan a large field-of-view without moving the sample. In addition, the LEAP workstation harnesses the highly accurate galvanometer mirrors to direct the rapid-pulse laser, which fires up to 1,000 pulses/second. Neighboring cells are virtually unaffected by elimination of the unwanted cells. Stem Cell Manager The LEAP Stem Cell Manager enables automation and management of stem cell colony purification and processing with a suite of applications: Stem Cell Passage, Embryoid Body (EB) Generation, and Stem Cell Colony Purification. With the Stem Cell Manager, researchers can physically passage ES and ips cell colonies in a way that is rapid, automated, and enzyme-free. Stem cell culture propagation is highly consistent, generating cultures with uniform and tunable colony sizes. The Stem Cell Manager also enables generation of specifically-sized EBs, significantly improving their differentiation yield into specialized cell types. LEAP Stem Cell Manager enhances stem cell research: Ensures rapid and consistent passage of stem cell cultures Achieves greater viability, yield, and genetic stability Enables greater yield and differentiation potential with uniform-size EBs Generates new and/or genetically modified stem cell lines Processes cells and cultures in a sterile, closed environment Embryoid Body Generation Application Standard methods for generating EBs result in heterogeneous EB populations that vary in size and morphology, and possess limited differentiation potential and low production yields. The EB Generation Application on the LEAP workstation provides an efficient, automated method for generating uniform EB populations of user-defined sizes, allowing high-yield preparation of EBs with the highest differentiation efficiency. Facilitating stem cell research by providing a reproducible method for production of high-quality, large-scale homogeneous EB cultures, LEAP provides clear advantages for the stem cell research lab. LEAP Cell Processing Workstation 3

4 Stem Cell Manager Applications Standardization and Reproducibility for Stem Cell Research Stem Cell Passage Application Enzymatic methods for stem cell passaging are widely used and easily Stem Cell Passage Application scaled up for large-scale expansion, but result in variable-sized colonies and significant cellular trauma. In contrast, non-enzymatic, physical passaging of stem cells better maintains genetic stability in long-term culture, decreases Generation of uniformly sized colonies compared to conventional enzymatic methods Uniformity of ipsc cultures after Passage cellular trauma, and is required for production of new GMP-quality, clinical- 250 Cells/Colony (#) grade stem cell lines. Until now, physical passaging of stem cell colonies has only been possible through manual methods, which are labor-, cost-, and timeintensive, and therefore not suitable for large-scale culture. The automated Stem Cell Passage Application on the LEAP workstation automates physical passaging of stem cells through laser processing, allowing the stem cell researcher to scale up physical passaging of stem cells and easily generate LEAP Collagenase Trypsin Passage Method the cells needed for the highest quality research. Stem Cell Colony Purification Application Stem cells can be difficult to purify in part due to their growth as tight colonies, low abundance, sensitivity to stress, and/or lack of highly specific identifying Stem Cell Colony Purification Application markers. Separation into individual cells often causes death or differentiation, whereas physical picking of colonies is slow, tedious, and can result in BEFORE AFTER contamination. The highly accurate targeting of the LEAP workstation s rapid laser enables purification of intact stem cell colonies right where they grow by eliminating all undesired cells in a well without significantly disturbing the desired stem cell colony. Live hipscs stained with TRA1-60. After purification, cells washed 1x with medium and fresh feeders added. Users can define regions to remove undesired cells around the colony or even in the middle of the colony. 4 LEAP Cell Processing Workstation *P<0.001

5 Adherent Cell Purification Applications Facilitating Efficient and Automatable Cell Biology Research Primary cells have the potential to provide more physiologically relevant The LEAP workstation overcomes these challenges in primary and adherent cell information than immortalized cell lines for discovery biology and drug purification by eliminating contaminant cells without disruption or manipulation discovery research. However, obtaining primary cell preparations of sufficient of the desired cells. Purification of adherent cells is easily automated, enabling purity, especially preparations of adherent cells, can be challenging due to the high reproducibility and productivity. Cell processing is fast elimination of low number of initial cells and the damaging methods used for purification. contaminants from a single well can take only seconds. Purification using the Most current approaches, such as flow cytometry or bead capture, require LEAP workstation causes less trauma to the remaining adherent cells than enzyme treatment to suspend cells for processing, altering cell morphology, other more invasive methods, as has been demonstrated by monitoring growth physiology, and gene expression, and decreasing cell survival and yield. rates, cloning efficiencies, viability, and proper stem cell gene expression and genetic stability over time. Because the desired cells never leave the well, yield is extremely high, even when working with small numbers of cells. Adherent Cell Purification Application Efficient purification of a pure population of adherent endothelial cells Impure preparation of primary rat brain microvascular endothelial cells stained with CellTracker Green and DiI-Ac-LDL (left). Contaminating fibroblasts are identified by arrows (upper inserts). Contaminating cells were destroyed using the 532 nm laser of LEAP, leaving behind a pure population of DiI-Ac-LDL-stained endothelial cells (center). After 72 hours, extensive outgrowth of a pure endothelial cell population was observed. LEAP Cell Processing Workstation 5

6 Using the LEAP Workstation Power, Precision, and Control The LEAP workstation brings automated cell processing and purification to research labs and core facilities working in a wide variety of cell-based LEAP Software systems, with both label-free and fluorescence applications. LEAP Software The LEAP workstation comes with elegant and intuitive software that enables reproducible generation of highly-pure, highly-homogenous stem cell and adherent cell cultures. Users choose from predefined applications that step through the appropriate parameter settings for the specific application. Real-time and Dynamic Image-based Gating The LEAP workstation software allows selection of cells through a wide variety of gating criteria shape, size, fluorescence intensity, and localization to identify cells of interest. Histogram, scatter plots, and meta-images display the actual cells being imaged and cells of interest are highlighted by colored overlays. Real-time imaging and gating of live cells means users can dynamically adjust gating parameters to explore how alternate gating criteria might improve or alter selection of cells of interest. Alternatively, the gates can be fixed for automated processing. 6 LEAP Cell Processing Workstation Novel software interface combines both image and flow cytometry features, allowing cell selection using a variety of criteria

7 LEAP Products and Services A Complement of Tools and Global Support C-Lect Consumables As a complement to the LEAP workstation, Cyntellect offers C-Lect About Cyntellect Consumables kits and tools optimized and validated for specific applications on LEAP. Each kit comes complete with reagents, C-lect Cyntellect is dedicated to setting new standards in cell analysis, purification, microplates and protocols. Kits are available for stem cell passage, EB and processing technology. Cyntellect s products support key applications generation, antibody secretion (human and murine), and cell purification. to advance life science research, biopharmaceutical production, stem cell The kits combine with ready-to-use software protocols available on the research, and drug discovery. The company s patented technology employs LEAP workstation to streamline your workflow. in situ, microplate-based cytometry to analyze cells with minimal sample manipulation and process cells with great precision and efficiency. Cyntellect s LEAP Workstation Global Support and Service expanding cellular analysis and processing portfolio is expected to play On-site installation and service is provided by experienced Field Service an enabling role in the coming age of advanced cell-based diagnostics, Engineers. Customer training is conducted on-site by our expert Field therapeutics, and regenerative medicine. Applications Scientists. Our global support network ensures all customers will receive quality training and service regardless of their geographic location. Applications support offers interactive web-based tutorials on the newest developments in cell-based assays. LEAP Cell Processing Workstation 7

8 US CORPORATE HEADQUARTERS 6620 Mesa Ridge Road San Diego, CA main fax US toll-free Cyntellect Europe, Ltd. Wales 1 Business Park Building 104, Ground Floor Newport Road Magor, UK NP26 3DG +44 (0) main +44 (0) fax Copyright 2010 by Cyntellect, Inc. All rights reserved. The LEAP workstation, software, and portions of this document may be protected by one or more patents and/or pending patent applications, including: United States: 5,874,266; 6,143,535; 6,514,722; 6,534,308; 6,642,018; 6,753,161; 6,804,385; 7,092,557; 7,129,070; 7,300,795; 7,425,426; 7,505,618; and 7,622,274. Australia: ; ; ; and France: and Germany: and Ireland: Netherlands: Sweden: United Kingdom: and Cyntellect reserves the right to make modifications and/or additions to the information in this document without notice. Cyntellect is a registered trademark. LEAP is a trademark of Cyntellect.