Applications & Publications
Technical Notes
Large Particle Flow Cytometry Allows High Quality Isolation of Viable Cardiomyocytes and Other Myocytes (QTN-028)
April 22, 2020
Comparison of Total Fluorescence in Cell Clusters versus Fluorescence of the Individual Cells in the Cluster (QTN-025)
February 16, 2017
Method for Analysis and Sorting of Live Adipocytes (QTN-022)
December 15, 2014
Analysis and sorting of human neurospheres based on size and optical density measurements (QTN-016)
December 01, 2011
The purpose of this experiment was to test the feasibility of using the COPAS PLUS instrument (Union Biometrica, Inc.) to analyze and sort human neurospheres based on size and optical density measurements and to determine the influence on viability and proliferation of the neurospheres post-sorting compared to manual sorting.
Analysis and sorting of Stem Cell Clusters and Adipocytes
Automated Analysis and Dispensing of Human Stem Cell Clusters and Adipocytes.
Use of the COPAS Select for the analysis and sorting of two different human cell samples: individual stem cell clusters (EBs) and human adipocytes. Cell aggregates and individual cells were accurately dispensed and visually inspected for structural integrity.
An Overview of COPAS™ Large Particle Flow Cytometry for the Analysis and Sorting of Large Cells and Cell Clusters including Stem Cells, Embryoid Bodies and Cardiomyocytes (QTN-013)
Automated Sorting of Adult Stem Cells and Mouse Embryonic Bodies (QTN-011)
Use of the instrument for analysis and sorting of individual and clusters of adult stem cells and embryoid bodies, based on both size and fluorescence intensity properties. (QTN-011)
Automated analysis and sorting of human induced pluripotent stem cell (hiPS) clusters using large particle flow cytometry. (QTN-018)
The purpose of this experiment was to test the feasibility of using the COPAS PLUS HTS instrument (Union Biometrica, Inc.) to analyze and sort intact human induced pluripotent stem cell (hiPS) clusters based on their size, optical density and fluorescent properties.
Automated Analysis and Sorting of Cardiomyocyte Stem Cell Clusters (QTN-001)
Use of the COPAS Select for the analysis and sorting of individual embryoid bodies (EBs) from cultured samples. Stem cell aggregates were accurately dispensed into multi-well plates and visually inspected for viability. (QTN-001)
Publications
Trajectory reconstruction identifies dysregulation of perinatal maturation programs in pluripotent stem cell-derived cardiomyocytes
Kannan et al. April 25, 2023 PMID: 37014753 PMCID: PMC10545814 DOI: 10.1016/j.celrep.2023.112330
Trajectory reconstruction identifies dysregulation of perinatal maturation programs in pluripotent stem cell-derived cardiomyocytes
Cell Cluster Sorting in Automated Differentiation of Patient-specific Induced Pluripotent Stem Cells Towards Blood Cells
Ma et al. May 17, 2022 Front. Bioeng. Biotechnol., 17 May 2022 | https://doi.org/10.3389/fbioe.2022.755983
Cell Cluster Sorting in Automated Differentiation of Patient-specific Induced Pluripotent Stem Cells Towards Blood Cells
Single-Cell Sorting of Non-human Primate Adipocytes with Large-particle Flow Cytometry
Robino et al. November 04, 2021 Current Protocols,1, e271. doi: 10.1002/cpz1.271
Single-Cell Sorting of Non-human Primate Adipocytes with Large-particle Flow Cytometry
New method of FACS analyzing and sorting of intact whole ovarian fragments (COPAS) after long time (24 h) cooling to 5 degrees C before cryopreservation
Wang et al. December 27, 2020 Cell Tissue Bank. https://doi.org/10.1007/s10561-020-09898-1
View AbstractNew method of FACS analyzing and sorting of intact whole ovarian fragments (COPAS) after long time (24 h) cooling to 5 degrees C before cryopreservation
As recently announced by the American Society for Reproductive Medicine (ASRM), human ovarian tissue cryopreservation is an established option for fertility preservation in prepubertal girls and young women undergoing gonadotoxic treatments for cancer as well as some autoimmune diseases. Proper ovarian tissue assessment before and after cryopreservation is essential to increase success rates. Ovarian fragments from 16 patients were divided into small pieces in form of cortex with medulla, and randomly divided into the following two groups. Pieces of Group 1 (n = 16) were frozen immediately after operation, thawed and just after thawing their quality was analyzed. Group 2 pieces (n = 16) after operation were cooled to 5 °C for 24 h, then frozen after 24 h pre-cooling to 5 °C, thawed and just after thawing their quality was analyzed. The effectiveness of the pre-freezing cooling of tissue was evaluated by the development and viability of follicles (Calcein-AM and Propidium Iodide) using complex object parametric analyzer and sorter machine (COPAS). Positive effect of cooling of cells to low supra-zero temperatures on their future development after re-warming has been observed. New flow cytometry- technique is suitable for the evaluation and sorting of cryopreserved whole human whole intact ovarian fragments. Long time (24 h) cooling of ovarian tissue to 5 °C before cryopreservation has a trend of a cell viability increasing.
Adipocyte Piezo1 Mediates Obesogenic Adipogenesis Through the FGF1/FGFR1 Signaling Pathway in Mice
ShengPeng Wang et al. May 08, 2020 Nat Commun. 11(1):2303. doi: 10.1038/s41467-020-16026-w.
Adipocyte Piezo1 Mediates Obesogenic Adipogenesis Through the FGF1/FGFR1 Signaling Pathway in Mice
Transcriptomic entropy quantifies cardiomyocyte maturation at single cell level
Kannan et al. April 03, 2020 PMID: 34534204 PMCID: PMC8448341 DOI: 10.1371/journal.pcbi.1009305
Transcriptomic entropy quantifies cardiomyocyte maturation at single cell level
PGC1/PPAR Drive Cardiomyocyte Maturation through Regulation of Yap1 and SF3B2
Murphy et al. February 07, 2020 bioRxiv preprint: doi: https://doi.org/10.1101/2020.02.06.937797
PGC1/PPAR Drive Cardiomyocyte Maturation through Regulation of Yap1 and SF3B2
Proteogenomic single cell analysis of skeletal muscle myocytes.
Fomchenko et al. January 24, 2020 bioRxiv preprint: doi: https://doi.org/10.1101/2020.01.23.916791
Proteogenomic single cell analysis of skeletal muscle myocytes.
Activation of invariant natural killer T cells stimulates adipose tissue remodeling via adipocyte death and birth in obesity
Park et al. December 01, 2019 Genes Dev. December 1, 2019 33: 1657-1672; Published in Advance November 14, 2019, doi:10.1101/gad.329557.119
Activation of invariant natural killer T cells stimulates adipose tissue remodeling via adipocyte death and birth in obesity
Large Particle Fluorescence-Activated Cell Sorting Enables High-Quality Single-Cell RNA Sequencing and Functional Analysis of Adult Cardiomyocytes
Kannan et al. August 15, 2019 PMID: 31415233 PMCID: PMC6699769 DOI: 10.1161/CIRCRESAHA.119.315493
Large Particle Fluorescence-Activated Cell Sorting Enables High-Quality Single-Cell RNA Sequencing and Functional Analysis of Adult Cardiomyocytes
Automation for analysis and handling of cell clusters, tumor spheres and organoid bodies
Pulak et al. February 03, 2018
View AbstractAutomation for analysis and handling of cell clusters, tumor spheres and organoid bodies
Analysis and sorting of cell clusters generated in Corning microcavity flasks.
Deciphering Cell Intrinsic Properties: A Key Issue for Robust Organoid Production
Picollet-D’hahan et al. September 17, 2017 Trends in Biotechnology. 2017 Nov;35(11):1035-1048. doi: 10.1016/j.tibtech.2017.08.003. Epub 2017 Sep 17.
Deciphering Cell Intrinsic Properties: A Key Issue for Robust Organoid Production
Endothelin-1 mediates natriuresis but not polyuria during vitamin D-induced acute hypercalcaemia
Tokonami et al. March 27, 2017 First published: 23 March 2017, DOI: 10.1113/JP273610
Endothelin-1 mediates natriuresis but not polyuria during vitamin D-induced acute hypercalcaemia
Fluid shear stress increases transepithelial transport of Ca2+ in ciliated distal convoluted and connecting tubule cells
Mohammed et al. January 18, 2017 Published online before print January 18, 2017, doi: 10.1096/fj.201600687RRR, May 2017, The FASEB Journal vol. 31 no. 5 1796-1806
Fluid shear stress increases transepithelial transport of Ca2+ in ciliated distal convoluted and connecting tubule cells
A 3D Toolbox to Enhance Physiological Relevance of Human Tissue Models
Picollet-D’hahan et al. August 04, 2016 Trends in Biotechnology. 2016 Sep;34(9):757-769. doi: 10.1016/j.tibtech.2016.06.012. Epub 2016 Aug 4.
A 3D Toolbox to Enhance Physiological Relevance of Human Tissue Models
Urinary ß-galactosidase stimulates Ca2+ transport by stabilizing TRPV5 at the plasma membrane
Leunissen et al January 07, 2016 Glycobiology, Volume 26, Issue 5, 1 May 2016, Pages 472–481, https://doi.org/10.1093/glycob/cwv172 , Published: 07 January 2016
Urinary ß-galactosidase stimulates Ca2+ transport by stabilizing TRPV5 at the plasma membrane
Controlled 3D culture in Matrigel microbeads to analyze clonal acinar development
Dolega et al. June 01, 2015 Biomaterials. 2015 Jun;52:347-57. doi: 10.1016/j.biomaterials.2015.02.042. Epub 2015 Mar 3.
Controlled 3D culture in Matrigel microbeads to analyze clonal acinar development
Protein Phosphatase 1 Inhibitor-1 Deficiency Reduces Phosphorylation of Renal NaCl Cotransporter and Causes Arterial Hypotension.
Nicolas Picard,*† Katja Trompf,* Chao-Ling Yang,‡ R. Lance Miller,§ Monique Carrel,*Dominique Loffing-Cueni,* Robert A. Fenton,|| David H. Ellison,‡ and Johannes Loffing* April 01, 2014 J Am Soc Nephrol 25: 511-522, 2014. doi: 10.1681/ASN.2012121202 *Institute of Anatomy, University of Zurch, Zurich, Switzerland †Paris Cardiovascular Research Center, Institut National de la Santé et de la Recherche Médicale Unit 970, Université Paris-Descartes, Paris, France; ‡Department of Medicine, Oregon Health and Science University, Portland, Oregon; §National Heart Lung Blood Institute, National Institutes of Health, Bethesda, Maryland; and || Department of Biomedicine, Aarhus, University, Aarhus, Denmark
View AbstractProtein Phosphatase 1 Inhibitor-1 Deficiency Reduces Phosphorylation of Renal NaCl Cotransporter and Causes Arterial Hypotension.
The thiazide-sensitive NaCl cotransporter (NCC) of the renal distal convoluted tubule (DCT) controls ion homeostasis and arterial BP. Loss-of-function mutations of NCC cause renal salt wasting with arterial hypotension (Gitelman syndrome). Conversely, mutations in the NCC-regulating WNK kinases or kelch-like 3 protein cause familial hyperkalemic hypertension. Here, we performed automated sorting of mouse DCTs and microarray analysis for comprehensive identification of novel DCT-enriched gene products, which may potentially regulate DCT and NCC function. This approach identified protein phosphatase 1 inhibitor-1 (I-1) as a DCT-enriched transcript, and immunohistochemistry revealed I-1 expression in mouse and human DCTs and thick ascending limbs. In heterologous expression systems, coexpression of NCC with I-1 increased thiazide-dependent Na(+) uptake, whereas RNAi-mediated knockdown of endogenous I-1 reduced NCC phosphorylation. Likewise, levels of phosphorylated NCC decreased by approximately 50% in I-1 (I-1(-/-)) knockout mice without changes in total NCC expression. The abundance and phosphorylation of other renal sodium-transporting proteins, including NaPi-IIa, NKCC2, and ENaC, did not change, although the abundance of pendrin increased in these mice. The abundance, phosphorylation, and subcellular localization of SPAK were similar in wild-type (WT) and I-1(-/-) mice. Compared with WT mice, I-1(-/-) mice exhibited significantly lower arterial BP but did not display other metabolic features of NCC dysregulation. Thus, I-1 is a DCT-enriched gene product that controls arterial BP, possibly through regulation of NCC activity.
AUTOMATED ANALYSIS AND SORTING OF HUMAN IPS CELL CLUSTERS BY LARGE PARTICLE FLOW CYTOMETRY
ISSCR 2013
Wanek, Paul¹, Smet, Francis², Haupt, Simone³, Peitz, Michael³, Bongaarts, Rico², Oliver Brüstle³, Pulak, Rock4, Zenke, Martin¹
June 13, 2013
AUTOMATED ANALYSIS AND SORTING OF HUMAN IPS CELL CLUSTERS BY LARGE PARTICLE FLOW CYTOMETRY
1) Institute for Biomedical Engineering, Department of Cell Biology, RWTH Aachen University Medical School, RWTH Aachen University, Aachen, Germany, 2) Union Biometrica, Geel, Belgium, 3) Institute of Reconstructive Neurobiology, University of Bonn, Bonn, Germany, 4) Union Biometrica, Holliston, MA, USA
Flow-based pipeline for systematic modulation and analysis of 3D tumor microenvironments.
Li CY, Wood DK, Huang JH, Bhatia SN.
Chemical Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, United States.
April 23, 2013
Lab Chip. 2013 Apr 23;13(10):1969-78. doi: 10.1039/c3lc41300d. Epub 2013 Apr 5.
Flow-based pipeline for systematic modulation and analysis of 3D tumor microenvironments.
A PRIMARY CULTURE OF DISTAL CONVOLUTED TUBULES EXPRESSING FUNCTIONAL THIAZIDE-SENSITIVE NaCl TRANSPORT.
Markadieu N, San-Cristobal P, Nair A, Lenssen E, Tudpor K, van Zeeland F, Bindels RJ, Hoenderop JG. July 03, 2012 Am J Physiol Renal Physiol. 2012 Jul 3.
View AbstractA PRIMARY CULTURE OF DISTAL CONVOLUTED TUBULES EXPRESSING FUNCTIONAL THIAZIDE-SENSITIVE NaCl TRANSPORT.
Radboud University Nijmegen Medical Centre.
Automated analysis and sorting of Human Induced Pluripotent Stem cells (HIPS) clusters using large particle Flow Cytometry
ISSCR 2012
Wanek, Paul², Smet, Francis¹, Peitz, Michael³, Bongaarts, Rico¹, Pulak, Rock4, Zenke, Martin²
June 14, 2012
Automated analysis and sorting of Human Induced Pluripotent Stem cells (HIPS) clusters using large particle Flow Cytometry
1) Union Biometrica, Geel, Belgium, 2) Institute for Biomedical Engineering, Universitätsklinikum Aachen, RWTH, Aachen, Germany, 3) Institute of Reconstructive Neurobiology, University of Bonn, Bonn, Germany, 4) Union Biometrica, Holliston, MA, USA
Automated neurosphere sorting and plating by the COPAS large particle sorter is a suitable method for high-throughput 3D in vitro applications
ISSCR 2012
Gassmann K.², Smet F.¹, Baumann J.², Giersiefer S.², Schreiber T.², Schuwald J.², R.Bongaarts², Fritsche E.²
June 14, 2012
Automated neurosphere sorting and plating by the COPAS large particle sorter is a suitable method for high-throughput 3D in vitro applications
2) Leibniz Research Institute for Environmental Medicine, Department Molecular Toxicology, Duesseldorf, Germany. 1) Union Biometrica, Geel, Belgium.
Automated neurosphere sorting and plating by the COPAS large particle sorter is a suitable method for high-throughput 3D in vitro applications
Gassmann K, Baumann J, Giersiefer S, Schuwald J, Schreiber T, Merk HF, Fritsche E. May 02, 2012 http://dx.doi.org/10.1016/j.tiv.2012.04.025; Toxicology in Vitro Volume 26, Issue 6, September 2012, Pages 993–1000
View AbstractAutomated neurosphere sorting and plating by the COPAS large particle sorter is a suitable method for high-throughput 3D in vitro applications
Leibniz Research Institute for Environmental Medicine, Department Molecular Toxicology, Auf'm Hennekamp 50, 40225 Duesseldorf, Germany.
Functional assessment of automatically sorted pancreatic islets using large particle flow cytometry.
Steffen A, Ludwig B, Krautz C, Bornstein S, Solimena M. September 01, 2011 Islets. 2011 Sep 1;3(5):267-70. Epub 2011 Sep 1.
View AbstractFunctional assessment of automatically sorted pancreatic islets using large particle flow cytometry.
Department of Medicine III; Technische Universität Dresden; Dresden, Germany.
Microbial encapsulation in monodisperse hydrogel microspheres enables fast and sensitive phenotypic analyses using flow cytometers
Lidia Delgado¹, Gloria Jurado², Gema Galayo², Elena Ogalla², Lourdes Moreno³, Juan C Rodríguez-Aguilera4, Ángel Cebolla5, Carolina Sousa³, María Flores² and Sebastián Chávez¹ February 28, 2011 Conference Presentation at: La Société Française de Microbiologie et l’Association Française de Cytométrie 1) Department of Genetics, Universidad de Sevilla, Seville, Spain. 2) Ingeniatrics Tecnologías SL, Spain. 3) Department of Microbiology and Parasitology, Universidad de Sevilla, Seville, Spain. 4) Centro Andaluz de Biología del Desarrollo, Universidad Pablo de Olavide, Seville, Spain. 5) Biomedal SL, Spain
Microbial encapsulation in monodisperse hydrogel microspheres enables fast and sensitive phenotypic analyses using flow cytometers
{gamma}-Adducin Stimulates the Thiazide-sensitive NaCl Cotransporter.
Dimke H, San-Cristobal P, de Graaf M, Lenders JW, Deinum J, Hoenderop JG, Bindels RJ. December 16, 2010 J Am Soc Nephrol. 2010 Dec 16.
View Abstract{gamma}-Adducin Stimulates the Thiazide-sensitive NaCl Cotransporter.
*Department of Physiology, Radboud University Nijmegen Medical Centre, Nijmegen, The Netherlands
[SA-PO2763] y-Adducin Functions as a Novel Regulator of the Thiazide-Sensitive NaCl-Cotransporter
Renal Week 2010
Henrik Dimke, Pedro San Cristobal, Mark J. J. De Graaf, Jacob (Jaap) Deinum, Jacques W. M. Lenders, Joost G. Hoenderop, Rene J. Bindels.
November 20, 2010
[SA-PO2763] y-Adducin Functions as a Novel Regulator of the Thiazide-Sensitive NaCl-Cotransporter
[F-PO1585] Distal Convoluted Tubule (DCT) Specific Transcripts Identified Using DCT Large Scale Sampling
Renal Week 2010
Nicolas Picard, R. Lance Miller, Johannes Loffing
November 19, 2010
[F-PO1585] Distal Convoluted Tubule (DCT) Specific Transcripts Identified Using DCT Large Scale Sampling
Transgenic Mice: Beyond the Knockout.
Miller RL. November 10, 2010 Am J Physiol Renal Physiol. 2010 Nov 10.
View AbstractTransgenic Mice: Beyond the Knockout.
1) National Institutes of Health.
Tissue engineered tumor models
M Ingram, GB Techy, BR Ward, SA Imam, R Atkinson, H Ho, CR Taylor August 01, 2010 Biotechnic & Histochemistry, August 2010, Vol. 85, No. 4 : Pages 213-229
Tissue engineered tumor models
Serum-free scaled up expansion and differentiation of murine embryonic stem cells to osteoblasts in suspension bioreactors.
Alfred R, Gareau T, Krawetz R, Rancourt D, Kallos MS. August 01, 2010 Biotechnol Bioeng. 2010 Aug 1;106(5):829-40.
View AbstractSerum-free scaled up expansion and differentiation of murine embryonic stem cells to osteoblasts in suspension bioreactors.
Pharmaceutical Production Research Facility (PPRF), Schulich School of Engineering, University of Calgary, 2500 University Drive N.W., Calgary, Alberta, Canada.
Reduction of diffusion barriers in isolated rat islets improves survival, but not insulin secretion or transplantation outcome
S Janette Williams,¹ Han-Hung Huang,¹ Karen Kover,³ Wayne Moore,³ Cory Berkland,² Milind Singh,² Irina V Smirnova,¹ Ronal MacGregor,4 and Lisa Stehno-Bittel¹ April 01, 2010 Organogenesis. 2010 Apr–Jun; 6(2): 115–124.
View AbstractReduction of diffusion barriers in isolated rat islets improves survival, but not insulin secretion or transplantation outcome
1Department of Physical Therapy and Rehabilitation Science; University of
4Department of Anatomy and Cell Biology; University of
2Department of Pharmeceutical Chemistry;
3Endocrinology Section;
Primary DCT Cell Culture Using Large Scale DCT Sampling from Mouse Kidneys
Katja Trompf,1 Nicolas Picard,1 Titia Woudenberg,2 Lance Miller,3 Rene J. Bindels,2 Hoenderop Joost,2 Johannes Loffing.1 November 06, 2009 Poster session: J Am Soc Nephrol 20: 2009 ; F-P01162
View AbstractPrimary DCT Cell Culture Using Large Scale DCT Sampling from Mouse Kidneys
1 Anatomy Inst., Univ.of Zurich, Switzerland; 2 Dpt of Physiology, Radboud Univ. Nijmegen MedicalCentre, Nijmegen, Netherlands; 3 Dpt of Pediatrics, Div. of Nephrology, School of Medicine, Univ. of Utah, Salt Lake City, UT.
Novel method for high-throughput colony PCR screening in nanoliter-reactors
Marcel Walser¹, Rene Pellaux¹, Andreas Meyer¹, Matthias Bechtold¹, Herve Vanderschuren², Richard Reinhardt³, Joseph Magyar4, Sven Panke¹ and Martin Held¹,* February 26, 2009 Nucl. Acids Res. (2009) 37 (8): e57/1-e57/8
View AbstractNovel method for high-throughput colony PCR screening in nanoliter-reactors
1) ETH Zurich, Institute of Process Engineering, BioProcess Laboratory (BPL), 2) ETH Zurich, Institute of Plant Science, Plant Biotechnology, Zurich, Switzerland, 3) Max Planck Institute for Molecular Genetics, Analytics & Computing, Berlin, Germany and 4) Harlan Laboratories Ltd., Environmental Safety and Metabolism, Itingen, Switzerland
Differentiation of COPAS-sorted non-endocrine pancreatic cells into insulin-positive cells in the mouse.
Kikugawa R, Katsuta H, Akashi T, Yatoh S, Weir GC, Sharma A, Bonner-Weir S January 30, 2009 Diabetologia. 2009 Jan 30. [Epub ahead of print: DOI 10.1007/s00125-009-1260-8]
View AbstractDifferentiation of COPAS-sorted non-endocrine pancreatic cells into insulin-positive cells in the mouse.
Section of Islet Transplantation and Cell Biology, Joslin Diabetes Center, 1 Joslin Place, Boston, MA, 02215, USA.
Impact of Islet Size on Graft Function
28th Workshop of the AIDPIT Study Group; 3rd European Diabetes Technology and Transplantation Meeting (EuDTT) – 2009, January 25-27
Lisa Stehno-Bittel, PhD.
January 25, 2009
Impact of Islet Size on Graft Function
1) University of Kansas Medical Center, Great Plain Diabetes Institute
Large Scale Sampling of Renal Distal Convoluted Tubules (DCT) by Complex Object Parametric Analysis and Sorting (COPAS)
Renal Week 2008: American Society of Nephrology (ASN) Annual Meeting, November 04 - 09, 2008; Philadelphia, Pennsylvania
Nicolas Picard* , Hannah Monyer, Beat Schwaller, R. Lance Miller, Johannes Loffing
November 04, 2008
Large Scale Sampling of Renal Distal Convoluted Tubules (DCT) by Complex Object Parametric Analysis and Sorting (COPAS)
Inst. of Anatomy, Univ. of Zurich, Zurich, Switzerland; Department of Clinical Neurobiology, Univ. of Heidelberg, Heidelberg, Germany; Unit of Anatomy, Dpt of Medicine, Univ. of Fribourg, Fribourg, Switzerland; Department of Pediatrics, Division of Nephrology, School of Medicine, Univ. of Utah, Salt Lake City, UT
Poster session: J Am Soc Nephrol 19: 2008 ; SA-PO2135
Abstract:
The renal DCT is important for the renal control of Na and K homeostasis. It also contributes to renal acid secretion and calcium and magnesium reabsorption. DCT function is difficult to study as the DCT is a rather short and difficult to identify nephron portion. Its hidden localization in the renal cortical labyrinth makes it also almost inaccessible to direct functional measurements. Recently, the group of R.L. Miller applied large-particle based flow cytometry (Complex Object Parametric Analysis and Sorting COPAS) (AJP Renal Physiol. 2006 Jul.) for large-scale sampling of collecting ducts (CD) using a transgenic mouse expressing GFP in CD. We used the same COPAS technique with a transgenic mouse expressing EGFP in the DCT in order to obtain large quantities of DCTs. Optimization of the COPAS sorting technique allows us to obtain up to 15000 fluorescent DCTs from one single mouse in less than three hours. Microscopical analysis revealed that almost all sorted tubules exhibit green fluorescence. Only occasionally some non-fluorescent tubule segments were attached. Western blot analysis confirmed the significant enrichment of the DCT-specific thiazide sensitive NaCl cotransporter in the sorted samples. Marker proteins for other nephron segments (i.e. NapiIIa - proximal tubule, NKCC2 thick ascending limb, AQP2 connecting tubule/collecting duct) were not detectable. Thus, COPAS allows rapid isolation of rather pure DCT preparations. These preparations will allow us now to study regulatory pathways involved in the DCT functions in vivo and to develop DCT cell culture for further functional testing in vitro.
Acute antibody-mediated complement activation mediates lysis of pancreatic islets cells and may cause tissue loss in clinical islet transplantation.
Tjernberg J, Ekdahl KN, Lambris JD, Korsgren O, Nilsson B. April 27, 2008 Transplantation. 2008 Apr 27;85(8):1193-9.
View AbstractAcute antibody-mediated complement activation mediates lysis of pancreatic islets cells and may cause tissue loss in clinical islet transplantation.
Department of Oncology, Rudbeck Laboratory, Uppsala, Sweden.
American Journal of Physiology: Renal Physiology, Automated Method for the Isolation of Collecting Ducts
R. Lance Miller1, Ping Zhang1, Tong Chen2, Andreas Rohrwasser2, and Raoul D. Nelson1* February 07, 2006 doi:10.1152/ajprenal.00273.2005
View AbstractAmerican Journal of Physiology: Renal Physiology, Automated Method for the Isolation of Collecting Ducts
1) Department of Pediatrics, Division of Nephrology, University of Utah, School of Medicine, Salt Lake City, UT, USA. 2) Department of Human Genetics, University of Utah, School of Medicine, Salt Lake City, UT, USA.
Validation of Large Particle Flow Cytometry for the Analysis and Sorting of Intact Pancreatic Islets.
Fernandez, Luis A.1; Hatch, Eric W.1; Armann, Barbara2; Odorico, Jon S.1; Hullett, Debra A.1; Sollinger, Hans W.1; Hanson, Matthew S.1 September 27, 2005 Transplantation. 80(6):729-737, September 27, 2005.
View AbstractValidation of Large Particle Flow Cytometry for the Analysis and Sorting of Intact Pancreatic Islets.
1) Department of Surgery, University of Wisconsin-Madison, Madison, WI. 2) Department of Surgery, University of Leipzig, Germany
Multicellular Cytometric Analyses and Sorting on Stem Cell Clusters and Embryonic Stem Cell Clusters (Poster)
3rd Annual ISSCR Meeting 2005, June 23 - 25
Tak Hung, Ph.D.1, Szczepan Baran, V.M.D.2, Carol Ware, Ph.D.2, Rock Pulak, Ph.D.1
June 23, 2005
Multicellular Cytometric Analyses and Sorting on Stem Cell Clusters and Embryonic Stem Cell Clusters (Poster)
1) Union Biometrica Inc., Holliston, MA 01746 USA; 2) Department of Comparative Medicine, University of Washington, Seattle, WA, USA
Large Particle Flow Cytometry as a New Automated Method of Islet Yield Determination (Poster PS-031)
International Pancreas and Islet Transplant Association (IPITA) 2005 Meeting, May 4-7, 2005
M. S. Hanson1, E. W. Hatch1, B. Armann1, D. A. Hullett1, J. S. Odorico1, H. W. Sollinger1, L. A. Fernandez1
May 04, 2005
Large Particle Flow Cytometry as a New Automated Method of Islet Yield Determination (Poster PS-031)
1Surgery, University of Wisconsin-Madison, Madison, WI, USA
Rapid and Sensitive Assessment of Human Islet Viability by Multiparameter Flow Cytometry on Intact and Dissociated Islets (Platform Session OP-139)
International Pancreas and Islet Transplant Association (IPITA) 2005 Meeting, May 4-7, 2005
M. S. Hanson1, B. Armann1, E. W. Hatch1, D. A. Hullett1, J. S. Odorico1, H. W. Sollinger1, L. A. Fernandez1
May 04, 2005
Rapid and Sensitive Assessment of Human Islet Viability by Multiparameter Flow Cytometry on Intact and Dissociated Islets (Platform Session OP-139)
1Surgery, University of Wisconsin-Madison, Madison, WI, USA
Flow cytometry for isolation and optical analysis of embryoid bodies from cultured mouse embryonic stem cells.
2nd Annual International Society of Stem Cell Researchers Meeting, June 10-13, 2004
Dr. H. Bohlen1, Dr. S Schwengberg1, R. Bongaarts2, Dr. R. Pulak3
June 10, 2004
Flow cytometry for isolation and optical analysis of embryoid bodies from cultured mouse embryonic stem cells.
1) Axiogenesis, Joseph-Stelzmann-Str. 50, Cologne, Germany, 2) Union Biometrica -Cipalstraat 3, Geel, Belgium, 3)Union Biometrica Inc., Holliston, MA 01746 USA
Automated fluorescence-activated collecting duct isolation for genomics and proteomics # 913.1.
Experimental Biology 2005, April 2 - 6, 2005
R. Lance Miller, Ping Zhang, Raoul D. Nelson.
Automated fluorescence-activated collecting duct isolation for genomics and proteomics # 913.1.
Pediatrics, University of Utah, 30 North 1900 East, SOM 4R312, Salt Lake City, Utah, 84108