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CellSys 100? Microdrop Maker,凝膠微滴法 (GMD),Gel Microdrop (GMD) Secretion

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產(chǎn)品名稱：CellSys 100? Microdrop Maker,凝膠微滴法 (GMD),Gel Microdrop (GMD) Secretion
產(chǎn)品型號：
產(chǎn)品展商：One Cell Systems ,onecell
產(chǎn)品文檔：無相關(guān)文檔

簡單介紹

One Cell Systems is developing and marketing a family of single cell assays for research and bioprocessing applications.

產(chǎn)品描述

The CellSys 100? Microdrop Maker has been engineered to maximize cell encapsulation while preserving cell integrity.

High rotation speeds and unique blade configuration allow the selection of microdrop diameter sizes between

10 and 100 microns. A schematic of the CellSys 100?
Microdrop Maker is shown below:

A Gel Microdrop (GMD) is a small, agarose-based microsphere that encapsulates an individual cell in a porous gel matrix.

By diffusing various reagents into the agarose matrix, one can perform a variety of assays on large populations of GMDs simultaneously. Cell phenotype and function can be examined on a cell-by-cell basisby flow cytometry because each cell is individually encapsulated. Viable cells can be sorted and recovered based on the results of these assays. Please click here for more information on FACS and GMDs.

106 cells can be encapsulated, assayed, sorted, and recovered in less than one day, making this technology a faster and more efficient alternative to limiting dilution cloning.

Characteristics of GMDs

1	GMDs are permeable to small molecules such as cell nutrients, cell culture media, and antibodies.
2	Cell viability, growth, and secretory activity are not significantly affected by encapsulation.
3	GMDs can be physically manipulated just like cells, i.e. they can be centrifuged, pipetted, incubated, and sorted by FACS.
4	The CellSys 100? Microdrop Maker is uniquely designed to generate GMDs between 10 and 100 microns in diameter, depending on the application and protocol used.

The Gel Microdrop (GMD) Secretion Assay rapidly identifies cells that secrete a protein of interest. Viable, high-secreting or rare cells can be isolated and recovered using fluorescence activated cell sorting (FACS). Please click here for more information on the GMD Protein Secretion Assay and FACS.

Encapsulated cells are analyzed or recovered using flow cytometry based on the amount of protein captured in each GMD. The capture and detection is analogous to the sandwich antibody format used in enzyme-linked immunosorbent assays (ELISA), however, the immunoreaction itself takes place within the agarose microsphere, leaving the encapsulated cell intact. A schematic representation of the GMD secretion assay is depicted below.

The GMD Secretion Assay Involves Four Easy Steps

1	Encapsulate individual cells in CelBioGel ? Encapsulation Matrix using the CellSys 100? Microdrop Maker.
2	Diffuse the biotinylated capture antibody into the GMDs. Streptavidin forms a bridge between the biotinylated encapsulation matrix and the biotinylated capture antibody.
3	Incubate the GMDs to allow the capture and retention of secreted protein within the agarose matrix. Label captured protein using fluorescently labeled reporter antibodies.
4	Identify and/or isolate secreting cells based on fluorescence intensity. GMD technology is compatible with fluorescence microscopy and flow cytometry methods.

Protein secreted by an individual cell is captured and fluorescently labeled within the GMD. Unoccupied GMDs and GMDs occupied by non-secreting cells do not fluoresce. The fluorescence intensity of each occupied GMD corresponds to the amount of protein secreted by the cell. Fluorescing GMDs occupied by secreting cells can be sorted and recovered based on level of protein secretion.

Advantages of the GMD Protein Secretion Assay

1	Enrich for stable, high producing clones in less than one day.
2	Isolate rare cells that occur at a frequency as low as 0.01%.
3	Eliminate time-consuming and laborious Limiting Dilution Cloning.
4	Improve yields and increase productivity.
5	Reduce development time and costs.

We offer a lease program which includes a CellSys 100? Microdrop Maker, CelMix? and CelBioGel? encapsulation reagents, training, and technical support. Alternatively, if you have a characterized cell line and two non-overlapping antibodies (one biotinylated and one fluorescently labeled), we will screen and sort your cells on a fee-for-service basis. Please contact us for details regarding either of these arrangements.

The Gel Microdrop (GMD) Growth Assay rapidly quantifies the effects of compounds such as antibiotics, cytotoxic agents, and growth factors on individual cell growth. Populations can be analyzed by flow cytometry to identify sub-populations of interest, such as those that are drug resistant.

The GMD Growth Assay Involves Three Easy Steps

1	Encapsulate cells in CelBiogel? Encapsulation Matrix using the CellSys 100? Microdrop Maker.
2	Culture the encapsulated cells in the presence of the compound of interest.
3	Fluorescently label the cells in order to analyze flow cytometry.

Individual cells are initially encapsulated in CelGel? Encapsulation Matrix. The resulting GMDs function as miniature Petri dishes. Subsequent cell growth and fluorescent labeling permit clonogenic growth to be quantified using flow cytometry.

Cells divide normally within GMDs. After staining with a specific marker, GMD fluorescence increases as cell number increases . GMDs containing clonogenic microcolonies can be analyzed or sorted based on the amount of cell growth.

Advantages of the GMD Growth Assay

1	Easily discriminate between rapidly and slowly growing cells.
2	Assay the effects of drugs at the individual cell level.
3	Dramatically reduce assay time.
4	Recover antibiotic resistant organisms.
5	Automate drug discovery processes using flow cytometry.
6	Sort and recover cells of interest to develop cell lines or to identify genes of interest.

The CellSys 100? Microdrop Maker, CelMix? and CelGel? Encapsulation reagents are available for purchase. Alternatively, we can screen your cells on a fee-for-service basis. Please contact us for details regarding either of these arrangements.

One Cell Systems Publications (papers & books)

Secretion

1	Kenney, J.S., et al., Production of monoclonal antibodies using a secretion capture report web. Biotechnology (N Y), 1995. 13(8): p. 787-90.
2	Gray, F., J.S. Kenney, and J.F. Dunne, Secretion capture and report web: use of affinity derivatized agarose microdroplets for the selection of hybridoma cells. J Immunol Methods, 1995. 182(2): p. 155-63.
3	Nilsson, K., et al., Production of monoclonal antibodies by agarose-entrapped hybridoma cells. Methods in Enzymology, 1996. 121.
4	Weaver, J.C., P. McGrath, and S. Adams, Gel microdrop technology for rapid isolation of rare and high producer cells. Nat Med, 1997. 3(5): p. 583-5.
5	Holmes, P. and M. Al-Rubeai, Improved cell line development by a high throughput affinity capture surface display technique to select for high secretors. J Immunol Methods, 1999. 230(1-2): p. 141-7.
6	Hammill, L., J. Welles, and G.R. Carson, The gel microdrop secretion assay: identification of a low productivity subpopulation arising during the production of human antibody in CHO cells. Cytotechnology, 2000. 34: p. 27-37.
7	Turcanu, V. and N.A. Williams, Cell identification and isolation on the basis of cytokine secretion: a novel tool for investigating immune responses. Nat Med, 2001. 7(3): p. 373-6.
8	Madlambayan, G.J., et al., Controlling culture dynamics for the expansion of hematopoietic stem cells. J Hematother Stem Cell Res, 2001. 10(4): p. 481-92.
9	Turcanu, V., T.R. Hirst, and N.A. Williams, Modulation of human monocytes by Escherichia coli heat-labile enterotoxin B-subunit; altered cytokine production and its functional consequences. Immunology, 2002. 106(3): p. 316-25.
10	Akselband, Y., J. Trnovsky, and P. McGrath, Transfected cell line enrichment using the Gel Microdrop (GMD) secretion assay. BioProcessing, 2003. 2(2): p. 83-88.
11	Akselband, Y., P. Moen, and P. McGrath, Isolation of rare isotype switch variants in hybridoma cell lines using an agarose gel microdrop-based protein secretion assay. ASSAY and Drug Develpment Technologies, 2003. 5: p. 619-626.
12	Atochina O., Mylvaganam R., Akselband Y., McGrath P., Comparison of results using the gel microdrop cytokine secretion assay with ELISPOT and intracellular cytokine staining assay. Cytokine, 2004. 27(4-5): p.120-8.

Growth

13	Weaver, J.C., et al., Gel microdrops: rapid detection and enumeration of individual microorganisms by their metabolic activity. Biotechnology (N Y), 1988. 6: p. 1084-1089.
14	Rosenbluh, A., et al., Cell-density-dependent lysis and sporulation of Myxococcus xanthus in agarose microbeads. Journal of Bacteriology, 1989(September): p. 4923-4929.
15	Williams, G.B., J.C. Weaver, and A.L. Demain, Rapid microbial detection and enumeration using gel microdroplets and colorimetric or fluorescence indicator systems. J Clin Microbiol, 1990. 28(5): p. 1002-8.
16	Nir, R., et al., Single-cell entrapment and microcolony development within uniform microsperes amenable to flow cytometry. Applied and Environmental Microbiology, 1990(September 1990): p. 2870-2875.
17	Nir, R., et al., Flow cytometry sorting of viable bacteria and yeasts according to b-galactosidase activity. Applied and Environmental Microbiology, 1990(December 1990): p. 3861-3866.
18	Weaver, J.C., et al., Rapid clonal growth measurements at the single-cell level: gel microdroplets and flow cytometry. Biotechnology (N Y), 1991. 9(9): p. 873-7.
19	Goguen, B. and N. Kedersha, Clonogenic cytotoxicity testing by microdrop encapsulation. Nature, 1993. 363(6425): p. 189-90.
20	Ryan, C., B.T. Nguyen, and S.J. Sullivan, Rapid assay for mycobacterial growth and antibiotic susceptibility using gel microdrop encapsulation. J Clin Microbiol, 1995. 33(7): p. 1720-6.
21	Gift, E.A., et al., FACS-based isolation of slowly growing cells: double encapsulation of yeast in gel microdrops. Nat Biotechnol, 1996. 14(7): p. 884-7.
22	Katsuragi, T., et al., Gel microdroplet technique leaving microorganisms alive for sorting by flow cytometry. J Microbiol Methods, 2000. 42(1): p. 81-6.
23	Gift, E.A. and J.C. Weaver, Simultaneous quantitative determination of electroporative molecular uptake and subsequent cell survival using gel microdrops and flow cytometry. Cytometry, 2000. 39(4): p. 243-9.
24	Manome, A., et al., Application of gel microdroplet and flow cytometry techniques to selective enrichment of non-growing bacterial cells. FEMS Microbiology Letters, 2001. 197: p. 29-33.
25	Bogen, K.T., et al., Gel microdrop flow cytometry assay for low-dose studies of chemical and radiation cytotoxicity. Toxicology, 2001. 160(1-3): p. 5-10.
26	Zengler, K., et al., Cultivating the uncultured. Proc Natl Acad Sci U S A, 2002. 99(24): p. 15681-6.
27	Akselband Y., Cabral C., Sharpio DS, McGrath P., Rapid mycobacteria drug susceptibility testing using gel microdrop (GMD) growth assay and flow cytometry. J Microbiol Methods, 2005. 62(2): p. 181-97.

In Situ Hybridization

28	Nguyen, B.T., et al., In situ hybridization to chromosomes stabilized in gel microdrops. Cytometry, 1995. 21(2): p. 111-9.

Other

29	Weaver, J.C., et al., Microdrop technology: a general method for separating cells by function and composition, in Methods: A Companion to Methods in Enzymology. 1991, Academic Press: San Diego. p. 234-247.
30	LaSalle, J.M., et al., T-cell presentation of antigen requires cell-to-cell contact for proliferation and anergy induction. Differential MHC requirements for superantigen and autoantigen. J Immunol, 1993. 151(2): p. 649-57.
31	Su, M.W., et al., Cognate peptide-induced destruction of CD8+ cytotoxic T lymphocytes is due to fratricide. J Immunol, 1993. 151(2): p. 658-67.
32	Cox, D.L., et al., Treponema pallidum in gel microdroplets: a novel strategy for investigation of treponemal molecular architecture. Molecular Microbiology, 1995: p. 1151-1164.

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CellSys 100? Microdrop Maker,凝膠微滴法 (GMD),Gel Microdrop (GMD) Secretion

One Cell Systems is developing and marketing a family of single cell assays for research and bioprocessing applications.