Application Note. Yeast Analysis with a life - dead staining kit (Yeast control - viability)

Size: px
Start display at page:

Download "Application Note. Yeast Analysis with a life - dead staining kit (Yeast control - viability)"

Transcription

1 Application Note Yeast Analysis with a life - dead staining kit (Yeast control - viability) (CyFlow Cube 8 and CyFlow Cube 6 with CyView TM version 1.5 CyFlow Space) Introduction The Partec Yeast control - Viability staining kit aims to determine the percentage of living yeast cells from a growing culture or from transport competent yeast cells intended to be used for cultivation at a later stage (income quality control of yeast cultures). The result of such a measurement is the total amount of yeast cell (cell concentration) and the ratio of living cells out of such a culture. Materials, Instrument and Methods Yeast cells: as a standard control please use fresh baker s yeast from the supermarket. Please do not use the dry yeast but the fresh, dehydrated package (maximum usage after purchase: for 10 days). Yeast control viability staining kit (product code ). The staining kit consists of 2 reagents: reagent A and reagent B. Additional material required but not provided by the staining kit: PBS The sample should be analyzed on a flow cytometer with a blue laser excitation light source (488 nm wavelength) capable of detecting FCS, SSC as well as green (FL1, interference bandpass filter 536 / 40 nm) and orange / red fluorescence (FL2, interference bandpass filter 590 / 50 nm or FL3, interference bandpass filter 675/20 depending on instrument type). The following examples will highlight measurements on a CyFlow Cube and a CyFlow space instrument. Reagent Specification Spectral characteristics Reagent A Live cell stain Comparable to Fluorescein isothiocyanate (FITC) Absorption max: 495nm Emission max: 519nm Reagent B Dead cell stain Table 1: Description of staining reagents Propidium iodide (PI) Absorption max: 530nm Emission max: 630nm 1/6

2 Sample preparation Cell suspension Starting with commercial dehydrated yeast: resuspend 0,01 g (10 mg) of yeast material in 10 ml PBS Starting with a yeast cell culture: take 10 µl of cells from a growing culture in mid-log phase and dilute with 990 µl PBS (dilution factor: 100) Staining reagents Take 1 ml of yeast suspension (either from the commercial dehydrated yeast or the growing yeast culture) and add 10 µl of reagent A. Incubate for 5 min at room temperature protected from light. Add 10 µl of reagent B and incubate for another 5 min at room temperature protected from light. The solution B contains propidium iodide and methanol and should be handled with care. Please follow the instructions given within the product leaflet. The sample is ready for analysis. Prepared samples should be analyzed within 15 min after preparation. Optional sample preparation: as a dead cell control prepare one lot of cells as described above and incubate the cells in an appropriate, heat-proof sample container at 70 C in a water bath for 10 min. In such way obtained cells can be mixed with fresh cells to generate a control sample consisting of living and dead cells. Analysis on a FCM Set-up of the instrument: 1. Select a layout with three or more plots (histogram and dotplots) showing at least FSC SSC and FL1 FL2 or FL3 (depends on the filter set-up) 2. Select FSC as trigger parameter LL = 100 log3 on FSC and SSC gain approximately 200 log4 on fluorescence parameters gain approximately Set the analysis speed to 1 µl / s 3. Attach a negative control sample without stain to fine adjust GAIN values and LL-value Observe the analysis rate (events / sec). If the value exceeds 1000 dilute the sample by a factor of 5 with PBS (and repeat again if the analysis speed is still too high). For later counting analysis keep rate / s below 1000! 2/6

3 Set the gain values of FSC and SSC in such a way that the cell population shows up in the center of the FSC SSC dotplot (fig. 1). Surround the yeast cells by a polygon and apply this polygon to the dot-plot FL1 vs. FL2/3. Set the gain values of FL detectors to display cells in the first of 4 log decades (fig. 1b) Yeast cells Background a) b) Fig. 1 FSC SSC plot allows identification of the yeast cells (a) which can be surrounded by a polygon and displayed in the fluorescence dot-plot (b) 4. Attach sample stained with FDA only (control), analyse and stop. Set compensation for FL1 vs FL2/3 Fig. 2 FDA vs PI plot (live vs dead) uncompensated without (a) and with compensation line for FDA vs FL3 (b) 5. Attach sample stained with PI only (control), analyse and stop. Set compensation for FL2/3 vs FL1. If required correct GAIN and LL values. 3/6

4 Fig. 3 FDA vs PI plot (live vs dead) uncompensated with compensation line for PI vs FL1 (a) and compensated (b) Sample analysis and counting: Analyse double stained samples. Use True Volumetric Counting to enumerate number of vital and dead cells (instructions below). Within the dotplot FL1 FL2/3 the fluorescence signals of vital and dead cells can be clearly identified (fig. 4) as vital cells are mainly green fluorescent whereas dead cells are mainly red fluorescent. Fig. 4 FDA vs PI plot (live vs dead) compensated without absolute counting results 4/6

5 Counting with Electrode Sensors (True Volumetric Absolute Counting, TVAC) In order to quantify the concentration of yeast cells (and vital cells) the measure mode Volumetric counting with electrodes should be used. Fill 850µl of sample in a standard sample tube. Mix well and attach it to the sample port. An analysis cycle according the below sequence is started automatically and a true absolute volumetric counting begins when the liquid level in the tube reaches the first, shorter sensor. 1. The system starts with an initialization sequence of Pre-run phase followed by 2. Stabilization switching into 3. Run phase (a pre-counting phase). When the liquid level in the sample tube reaches the start electrode the counting procedure initiates and the system switches from Run to 4. Count phase leading to the end of counting when the liquid level reached the second sensor 5. During Repositioning sample pump is moving into start position followed by 6. Clean which washes the sample port and flow cell 7. Ready is displayed when the system is ready to analyse the next sample The current phase is indicated always in the Instrument Settings Box next to the flow counter. With sample speed of 3 µl/s this procedure takes about 100 seconds. In order to minimize effects of particle sedimentation during the process it is recommended to use high speed values of 3-5 µl/s! The measured volume is fixed to 200 µl by the position of the electrodes. Start Count Stop 5/6

6 Cleaning and Shutdown process set speed = 4 µl/s o o Connect a sample tube filled with 1.5ml 0.5 % Hypochloride (Order No ) to disinfect the tubing run until system stops Connect a sample tube filled with 1.5ml Cleaning Solution (green solution, Order No ) run until system stops o Connect a sample tube filled with 1.5ml Decontamination Solution (violet solution, Order No ) run until system stops o Connect a sample tube filled with 1.5ml Sheath fluid (Order No ) run 2 minutes and STOP - Shut down FloMax - Switch off CyFlow Space instrument - Shut off the computer 6/6

CyFlow Cube series Appealing from every angle

CyFlow Cube series Appealing from every angle CyFlow Cube series Appealing from every angle www.sysmex-partec.com CyFlow Cube 6 and Cube 8: compact, economic flow cytometers with a great performance Panta rhei a flexible solution for demands in flow

More information

Evaluation of the CyFlow Cube 6 flow cytometer unit for the assessment of brewing yeast viability and vitality

Evaluation of the CyFlow Cube 6 flow cytometer unit for the assessment of brewing yeast viability and vitality Sysmex UK Ltd. Evaluation of the CyFlow Cube 6 flow cytometer unit for the assessment of brewing yeast viability and vitality 1 Evaluation of the Sysmex Cube 6 flow cytometer for the assessment of brewing

More information

Flow Cytometry - The Essentials

Flow Cytometry - The Essentials Flow Cytometry - The Essentials Pocket Guide to Flow Cytometry: 1. Know your Cytometer 2. Understanding Fluorescence and Fluorophores 3. Gating Process 4. Controls 5. Optimization 6. Panel Building 7.

More information

Spherotech, Inc. 1. SPHERO TM Technical Note STN-8 Rev C

Spherotech, Inc. 1. SPHERO TM Technical Note STN-8 Rev C SPHERO TM Technical Note STN- Rev C. 0070 CALIBRATION AND PERFORMANCE TRACKING OF FLOW CYTOMETERS USING SPHERO TM CALIBRATION PARTICLES Introduction The SPHERO TM Calibration Particles are versatile, stable,

More information

ab Glucose Uptake Assay Kit (Cell-based)

ab Glucose Uptake Assay Kit (Cell-based) ab204702 Glucose Uptake Assay Kit (Cell-based) Instructions for Use For measuring glucose uptake via flow cytometry and fluorescent microscopy. This product is for research use only and is not intended

More information

High-throughput automation with the Attune NxT Autosampler: consistent results across all wells and across plates

High-throughput automation with the Attune NxT Autosampler: consistent results across all wells and across plates APPLICATION NOTE Attune NxT Flow Cytometer with Autosampler High-throughput automation with the Attune NxT Autosampler: consistent results across all wells and across plates Introduction The emerging field

More information

ab Propidium Iodide Flow Cytometry Kit for Cell Cycle Analysis

ab Propidium Iodide Flow Cytometry Kit for Cell Cycle Analysis ab139418 Propidium Iodide Flow Cytometry Kit for Cell Cycle Analysis Instructions for Use To determine cell cycle status in tissue culture cell lines by measuring DNA content using a flow cytometer. This

More information

ab Propidium Iodide Flow Cytometry Kit for Cell Cycle Analysis

ab Propidium Iodide Flow Cytometry Kit for Cell Cycle Analysis ab139418 Propidium Iodide Flow Cytometry Kit for Cell Cycle Analysis Instructions for Use To determine cell cycle status in tissue culture cell lines by measuring DNA content using a flow cytometer. This

More information

CyFlow Space Your flexible flow cytometer

CyFlow Space Your flexible flow cytometer CyFlow Space Your flexible flow cytometer www.sysmex-partec.com CyFlow Space its flexibility gives you the space you need for your work Analysing cells and particles, be it from blood, plasma, tissue,

More information

CyFlow Space Your flexible flow cytometer

CyFlow Space Your flexible flow cytometer CyFlow Space Your flexible flow cytometer www.sysmex-partec.com CyFlow Space its flexibility gives you the space you need for your work Analysing cells and particles, be it from blood, plasma, tissue,

More information

ANAT 3231 Cell Biology Lab12 Stem Cell Analysis

ANAT 3231 Cell Biology Lab12 Stem Cell Analysis ANAT 3231 Cell Biology Lab12 Stem Cell Analysis 2 June 2010 Dr Antonio Lee Neuromuscular & Regenera9ve Medicine Unit School of Medical Sciences, UNSW Introduction to Flow Cytometry Contributed by Vittoria

More information

Institute of Science & Technology in Medicine Control of Substance Hazardous to Health COSHH assessment

Institute of Science & Technology in Medicine Control of Substance Hazardous to Health COSHH assessment Institute of Science & Technology in Medicine Control of Substance Hazardous to Health COSHH assessment Title of Experiment/Procedure: Cell counting and viability using FACS, counting beads and propidium

More information

Protocol for FACS analysis of HeLa cell transfectants

Protocol for FACS analysis of HeLa cell transfectants Protocol for FACS analysis of HeLa cell transfectants You can refer to: Marks et al., 1995, J. Cell Biol. 131: 351-369; Voorhees et al., 1995, EMBO J. 14: 4961-4975; or Marks et al., 1996, J. Cell Biol.

More information

11/19/2013. Janine Zankl FACS Core Facility 13. November Cellular Parameters. Cellular Parameters. Monocytes. Granulocytes.

11/19/2013. Janine Zankl FACS Core Facility 13. November Cellular Parameters. Cellular Parameters. Monocytes. Granulocytes. DEPARTEMENT BIOZENTRUM Janine Zankl FACS Core Facility 13. November 2013 Cellular Parameters Granulocytes Monocytes Basophils Neutrophils Lymphocytes Eosinophils Cellular Parameters 1 What Is Flow Cytometry?

More information

CyFlow Cube 6. Immunology Microbiology Industrial Applications Agrosciences Aquaculture. Flow Cytometry: Simply elegant. Elegantly simple.

CyFlow Cube 6. Immunology Microbiology Industrial Applications Agrosciences Aquaculture. Flow Cytometry: Simply elegant. Elegantly simple. CyFlow Cube 6 Immunology Microbiology Industrial Applications Agrosciences Aquaculture Flow Cytometry: Simply elegant. Elegantly simple. 01 Applications for Indus trial ApplicationS for Health care Agro

More information

Each question may have MULTIPLE correct answers. Select all that are correct.

Each question may have MULTIPLE correct answers. Select all that are correct. Knowledge Assessment Flow Cytometry Workshop, Part 1 April 20, 2015 Each question may have MULTIPLE correct answers. Select all that are correct. 1. Tandem dyes are a. highly stable fluorophores after

More information

CyFlow Space. Your Flexible Flow Cytometer.

CyFlow Space. Your Flexible Flow Cytometer. CyFlow Space Your Flexible Flow Cytometer www.sysmex-partec.com Ultimate Flexibility Modular System The CyFlow Space flow cytometer is a modular system with ultimate flexibility: from a basic configuration

More information

Cell Cycle Phase Determination Kit

Cell Cycle Phase Determination Kit Cell Cycle Phase Determination Kit Catalog Number KA1301 100 assays Version: 04 Intended for research use only www.abnova.com Table of Contents Introduction... 3 Intended Use... 3 Background... 3 General

More information

seminar

seminar seminar 02.26.-28.2013. Flow cytometry Process or measurement method can measure discrete properties physical, chemical, biochemical, biological parameters of separate particles, e.g. biological cells

More information

Definitions. What functions does a flow cytometer be able to do? The build of the Flow cytometry and sorting. Flow cytometry

Definitions. What functions does a flow cytometer be able to do? The build of the Flow cytometry and sorting. Flow cytometry The build of the Flow cytometry and sorting Flow cytometry Seminar Mónika Tóth 11-12-13.04.2011 Definitions Flow cytometry Process or measurement method can measure discrete properties physical, chemical,

More information

Flowcytometry Dirk Pacholsky

Flowcytometry Dirk Pacholsky Flowcytometry Dirk Pacholsky Flowcytometry: Overview 1 2 Overview Flow Cyto Metry Fluid Cell Measurement measuring cell properties of cells in suspension Most Flow Cytometer have Spatially separated Lasers

More information

BASICS OF FLOW CYTOMETRY

BASICS OF FLOW CYTOMETRY BASICS OF FLOW CYTOMETRY AUTHOR: Ana Isabel Vieira APPROVAL: Henrique Veiga Fernandes Ana Sílvia Gonçalves SOP.UCF.002 03-09-2015 Pag. 1/9 Overview Flow: Fluid Cyto: Cell Metry: Measurement Flow cytometry

More information

INTRODUCTION TO FLOW CYTOMETRY

INTRODUCTION TO FLOW CYTOMETRY DEPARTEMENT BIOZENTRUM INTRODUCTION TO FLOW CYTOMETRY F ACS C ore F acility Janine Zankl FACS Core Facility 3. Dezember 2015, 4pm Cellular Parameters Granulocytes Monocytes Basophils Lymphocytes Neutrophils

More information

Application Information Bulletin: Set-Up of the CytoFLEX Set-Up of the CytoFLEX* for Extracellular Vesicle Measurement

Application Information Bulletin: Set-Up of the CytoFLEX Set-Up of the CytoFLEX* for Extracellular Vesicle Measurement Application Information Bulletin: Set-Up of the CytoFLEX Set-Up of the CytoFLEX* for Extracellular Vesicle Measurement Andreas Spittler, MD, Associate Professor for Pathophysiology, Medical University

More information

InhibiScreen Kinase Inhibitor Assay

InhibiScreen Kinase Inhibitor Assay InhibiScreen Kinase Inhibitor Assay Flow Cytometric assessment of efficacy of PI3Kγ, PI3Kδ, BTK and SYK pathway inhibitors using the measurement of basophil activation in human EDTA and Heparin Whole Blood.

More information

AutoMACS Pro Presentation. Lausanne, EPFL-SV-PTECH-PTCF Alain Hirschy Application Specialist DACH

AutoMACS Pro Presentation. Lausanne, EPFL-SV-PTECH-PTCF Alain Hirschy Application Specialist DACH AutoMACS Pro Presentation Lausanne, EPFL-SV-PTECH-PTCF 9.6.2010 Alain Hirschy Application Specialist DACH Alainh@miltenyibiotec.de Agenda 1. Introduction to MACS Technology 2. Key componentsof the AutoMACS

More information

Attune TM Acoustic Focusing Cytometer Training. Manik Punj Attune Training

Attune TM Acoustic Focusing Cytometer Training. Manik Punj Attune Training Attune TM Acoustic Focusing Cytometer Training Manik Punj Attune Training Attune Training Agenda Section 1 An Introduction to Flow Cytometry Section 2 An Introduction to Acoustic Focusing Hydrodynamic

More information

Principles of flow cytometry: overview of flow cytometry and its uses for cell analysis and sorting. Shoreline Community College BIOL 288

Principles of flow cytometry: overview of flow cytometry and its uses for cell analysis and sorting. Shoreline Community College BIOL 288 Principles of flow cytometry: overview of flow cytometry and its uses for cell analysis and sorting Shoreline Community College BIOL 288 Flow Cytometry What is Flow Cytometry? Measurement of cells or particles

More information

Propidium Iodide. Catalog Number: Structure: Molecular Formula: C 27H 34I 2N 4. Molecular Weight: CAS #

Propidium Iodide. Catalog Number: Structure: Molecular Formula: C 27H 34I 2N 4. Molecular Weight: CAS # Catalog Number: 195458 Propidium Iodide Structure: Molecular Formula: C 27H 34I 2N 4 Molecular Weight: 668.45 CAS # 25535-16-4 Physical Description: Dark red crystals Description: Reagent used for the

More information

determine optimum instrument settings for their own instruments and establish their own daily values.

determine optimum instrument settings for their own instruments and establish their own daily values. PC7 (770/488) SETUP KIT 6607121 PN 4299504-C FLOW CYTOMETER ALIGNMENT VERIFICATION FLUOROSPHERES FLOW CYTOMETER DETECTOR STANDARDIZATION FLUOROSPHERES INTENDED USE For Research Use Only. Not for use in

More information

PowERFUL. INTUITIvE. customizable. ThE NEw STAR of. BENchToP FLow cytometry

PowERFUL. INTUITIvE. customizable. ThE NEw STAR of. BENchToP FLow cytometry PowERFUL. INTUITIvE. customizable. ThE NEw STAR of BENchToP FLow cytometry ABoUT AcEA Established in 2002, ACEA Biosciences, Inc. develops cutting-edge cell analysis platforms for life science research.

More information

Live and Dead Cell Assay

Live and Dead Cell Assay ab115347 Live and Dead Cell Assay Instructions for Use Differential fluorescent labeling of live and dead cells This product is for research use only and is not intended for diagnostic use. Last Updated

More information

NM interference in the flow cytometric Annexin V/Propidium Iodide measurement

NM interference in the flow cytometric Annexin V/Propidium Iodide measurement Project: VIGO NM interference in the flow cytometric Annexin V/Propidium Iodide measurement AUTHORED BY: DATE: Cordula Hirsch 20.01.2014 REVIEWED BY: DATE: Harald Krug 10.04.2014 APPROVED BY: DATE: DOCUMENT

More information

Attune NxT Acoustic Focusing Cytometer The next generation in acoustic cytometry

Attune NxT Acoustic Focusing Cytometer The next generation in acoustic cytometry Attune NxT Acoustic Focusing Cytometer The next generation in acoustic cytometry Maybelline Giam Field Application Scientist The world leader in serving science Attune NxT Flow Cytometer Attune NxT Acoustic

More information

Flow Cytometry for Ploidy and Genome Size analysis - 1. Functional principles

Flow Cytometry for Ploidy and Genome Size analysis - 1. Functional principles Flow Cytometry for Ploidy and Genome Size analysis - 1. Functional principles The technique of plant ploidy and genome size analysis in a flow cytometer is based on the fluorescent labelling of the nuclear

More information

Anatomy of a flow cytometer

Anatomy of a flow cytometer Anatomy of a flow cytometer Fluidics Optics Electronics Cells in suspension flow in single-file through an illuminated volume where they scatter light and emit fluorescence that is collected, filtered,

More information

Your Research, Revolutionized

Your Research, Revolutionized Your Research, Revolutionized Drive Your Research Forward Your research needs are evolving and with the CytoFLEX flow cytometer you ll see just how far your data can take you. CytoFLEX has the advanced

More information

FLOW CYTOMETRY. CyAn ADP. Analyzer

FLOW CYTOMETRY. CyAn ADP. Analyzer FLOW CYTOMETRY CyAn ADP Analyzer Experience the Power of the CyAn ADP and its optimal performance The Power of Detection The Power of Speed The Power of Ease The CyAn ADP Analyzer is the next step in Advanced

More information

Calcein AM Cell Viability Kit

Calcein AM Cell Viability Kit Instructions For Research Use Only. Not For Use In Diagnostic Procedures Calcein AM Cell Viability Kit Catalog# 4892-010-K 1000 Tests* * Calculated based on using 1 μm final concentration of Calcein AM;

More information

Phagocytosis Assay Kit (IgG PE)

Phagocytosis Assay Kit (IgG PE) Phagocytosis Assay Kit (IgG PE) Item No. 600540 www.caymanchem.com Customer Service 800.364.9897 Technical Support 888.526.5351 1180 E. Ellsworth Rd Ann Arbor, MI USA TABLE OF CONTENTS GENERAL INFORMATION

More information

A protocol for the determination of total cell concentration of natural microbial communities in drinking water with FCM

A protocol for the determination of total cell concentration of natural microbial communities in drinking water with FCM Techneau June 2007 A protocol for the determination of total cell concentration of natural microbial communities in drinking water with FCM Deliverable 3.3.7 Techneau June 2007 A protocol for the determination

More information

EdU Flow Cytometry Kit. User Manual

EdU Flow Cytometry Kit. User Manual User Manual Ordering information: (for detailed kit content see Table 2) EdU Flow Cytometry Kits for 50 assays: Product number EdU Used fluorescent dye BCK-FC488-50 10 mg 6-FAM Azide BCK-FC555-50 10 mg

More information

The Worlds First Smart Flow Cytometer

The Worlds First Smart Flow Cytometer A Flow Revolution. I N T R O D U C I N G The Worlds First Smart Flow Cytometer & Dual Use Flow Cell Cassette. ORFLO has done it again. Flow cytometry has just been revolutionized. By integrating fluorescence

More information

ab JC-10 Mitochondrial Membrane Potential Assay Kit Flow Cytometry

ab JC-10 Mitochondrial Membrane Potential Assay Kit Flow Cytometry ab112133 JC-10 Mitochondrial Membrane Potential Assay Kit Flow Instructions for Use For detecting mitochondrial membrane potential changes in cells using our proprietary fluorescence probe. This product

More information

ACTG Laboratory Technologist Committee Revised Version 2.0 ACTG Lab Man CD38 Quantitation 07 May 2004

ACTG Laboratory Technologist Committee Revised Version 2.0 ACTG Lab Man CD38 Quantitation 07 May 2004 1. Principle Flow Cytometric quantitation of CD38 expression on CD8+ T lymphocytes The purpose of this assay is to quantitate the surface expression of the CD38 molecule on CD8 positive T lymphocytes.

More information

Detection of bacterial aggregation by flow cytometry

Detection of bacterial aggregation by flow cytometry Detection of bacterial aggregation by flow cytometry Jack Coleman 1 and Melis McHenry 2 1 Enzo Life Sciences, Farmingdale NY, USA, 2 Beckman Coulter Inc., Miami FL, USA PROTEOSTAT Aggresome Detection Kit

More information

PERFECT-COUNT MICROSPHERES

PERFECT-COUNT MICROSPHERES PERFECT-COUNT MICROSPHERES Perfect-Count Microspheres-Product code PCB-100 for 100 tests Introduction In recent years, the determination of absolute cell counts has been shown to be relevant in different

More information

APO-DIRECT Kit. A Complete Kit for Measuring Apoptosis by Flow Cytometry. Cat. No. TNB-6611-KIT. Apoptotic Cells. Gate. S-phase. Apoptosis.

APO-DIRECT Kit. A Complete Kit for Measuring Apoptosis by Flow Cytometry. Cat. No. TNB-6611-KIT. Apoptotic Cells. Gate. S-phase. Apoptosis. APO-DIRECT Kit Cat. No. TNB-6611-KIT A Complete Kit for Measuring Apoptosis by Flow Cytometry Gate S-phase DNA Width Apoptosis G1 G2 DNA Area DNA Content Apoptotic Cells G1 S G2 DNA Content Apoptosis Technical

More information

Propidium Iodide Solution

Propidium Iodide Solution G-Biosciences 1-800-628-7730 1-314-991-6034 technical@gbiosciences.com A Geno Technology, Inc. (USA) brand name Propidium Iodide Solution (1mg/ml in deionized water) (Cat. # 786 1272, 786 1273) think proteins!

More information

Flow Cytometry For New PhDs

Flow Cytometry For New PhDs Flow Cytometry For New PhDs 2012 Simon Monard SCRM and CIR smonard@staffmail.ed.ac.uk Derek Davies CRUK derek.davies@cancer.org.uk Monday 20th and Tues 21st Feb 2012 Program Day 1: Day 2: Basics of flow

More information

SOPVII-7. Panel X: NK-characterization

SOPVII-7. Panel X: NK-characterization Created by judith.eckl Page 1 of 8 09/06/2011 SOPVII-7 Panel X: NK-characterization Date: Author: Petra Prinz, Judith Eckl Experimenter: Date: 08/06/2011 Experiment description: Version: 1.0 Start: End:

More information

EdU Click FC ROTI kit for Flow Cytometry

EdU Click FC ROTI kit for Flow Cytometry USER MANUAL EdU Click FC EdU Click FC Introduction and product description: The detection of cell proliferation is of utmost importance for assessing cell health, determining genotoxicity or evaluating

More information

Introduction to. BD FACSAria TM Cell Sorter. Flow = Fluid Cyto = Cell Metry = Measurement

Introduction to. BD FACSAria TM Cell Sorter. Flow = Fluid Cyto = Cell Metry = Measurement What is Flow Cytometry? Introduction to BD FACSAria TM Cell Sorter Flow = Fluid Cyto = Cell Metry = Measurement BD Biosciences Application Specialist 產品應用專員 Daisy Kuo 郭正佼 A variety of measurements are

More information

Methodology. Protocol for plant genome size estimation by flow cytometry (2) GSAD: a genome size in the Asteraceae database

Methodology. Protocol for plant genome size estimation by flow cytometry (2) GSAD: a genome size in the Asteraceae database Methodology Isolation of plant nuclei We recommend (unless specified otherwise) working under cold conditions (i.e. keep all solutions, buffers and prepared samples for analysis on ice, and do the chopping

More information

Establishing sirna assays in primary human peripheral blood lymphocytes

Establishing sirna assays in primary human peripheral blood lymphocytes page 1 of 7 Establishing sirna assays in primary human peripheral blood lymphocytes This protocol is designed to establish sirna assays in primary human peripheral blood lymphocytes using the Nucleofector

More information

igem 2018 InterLab Study Protocol Before You Begin

igem 2018 InterLab Study Protocol Before You Begin igem 2018 InterLab Study Protocol Before You Begin Read through this entire protocol sheet carefully before you start your experiment and prepare any materials you may need. In order to improve reproducibility,

More information

Detecting human circulating endothelial cells using the Attune Acoustic Focusing Cytometer

Detecting human circulating endothelial cells using the Attune Acoustic Focusing Cytometer APPLICATION NOTE Attune Acoustic Focusing Cytometer Detecting human circulating endothelial cells using the Attune Acoustic Focusing Cytometer Circulating endothelial cells (CECs) are mature cells shed

More information

ab Cell Viability Assay Kit Fluorometric Dual Green/Red

ab Cell Viability Assay Kit Fluorometric Dual Green/Red ab112121 Cell Viability Assay Kit Fluorometric Dual Green/Red Instructions for Use For detecting cell viability in suspension and adherent cells by using dual proprietary green and red fluorescence probes.

More information

Titration of Fluorochrome-Conjugated Antibodies for Labeling Cell Surface Markers on Live Cells

Titration of Fluorochrome-Conjugated Antibodies for Labeling Cell Surface Markers on Live Cells Titration of Fluorochrome-Conjugated Antibodies for Labeling Cell Surface Markers on Live Cells Ruud Hulspas 1 UNIT 6.29 1 Cytonome/ST, Boston, Massachusetts ABSTRACT Nonspecific antibody binding is best

More information

Cellometer. for Cell Counting & Analysis. Brewing Yeast Wine Yeast Platelets and Other Small Cells. Image Cytometer

Cellometer. for Cell Counting & Analysis. Brewing Yeast Wine Yeast Platelets and Other Small Cells. Image Cytometer Cellometer 2 Image Cytometer for Cell Counting & Analysis Brewing Yeast Wine Yeast and Other Small Cells Cellometer 2 Image Cytometer Optimized Analysis for Yeast and other Small Cells Features of the

More information

Welcome to the CORES Flow Cytometry Sorter Facility

Welcome to the CORES Flow Cytometry Sorter Facility Welcome to the CORES Flow Cytometry Sorter Facility The FACSAria I and FACSARIA III are twin laser high speed sorters capable of analyzing cells based on 7-8 distinct fluorescent properties (besides FSC

More information

MoxiCyte Viability Kit (#MXA055) User s Guide (Rev )

MoxiCyte Viability Kit (#MXA055) User s Guide (Rev ) MoxiCyte Viability Kit (#MXA055) User s Guide (Rev 20150406) FOR RESEARCH USE ONLY on the Moxi Flow or Zepi Flow Instruments Orflo Technologies, LLC Ketchum, ID Tech_support@orflo.com 1-855- 879-6694 Overview

More information

RayBio Apoptosis Detection Kit, MitoCapture

RayBio Apoptosis Detection Kit, MitoCapture RayBio Apoptosis Detection Kit, MitoCapture User Manual Version 1.0 Mar 17, 2015 RayBio Apoptosis Detection Kit, MitroCapture Protocol (Cat#: 68MC-Apo-S100) RayBiotech, Inc. We Provide You With Excellent

More information

SOPVII-7. Panel VII: Treg-FACS-staining

SOPVII-7. Panel VII: Treg-FACS-staining Created by judith.eckl Page 1 of 6 09/06/2011 SOPVII-7 Panel VII: Treg-FCS-staining Date: uthor: Judith Eckl Experimenter: Date: 08/05/2011 Experiment description: Version: 1.0 Start: End: ackground information:

More information

Autophagy/Cytotoxicity Dual Staining Kit

Autophagy/Cytotoxicity Dual Staining Kit Autophagy/Cytotoxicity Dual Staining Kit Catalog Number KA1299 96 assays Version: 05 Intended for research use only www.abnova.com Table of Contents Introduction... 3 Background... 3 Principle of the Assay...

More information

Multidrug Resistance Assay Kit (Calcein AM)

Multidrug Resistance Assay Kit (Calcein AM) Multidrug Resistance Assay Kit (Calcein AM) Item No. 600370 www.caymanchem.com Customer Service 800.364.9897 Technical Support 888.526.5351 1180 E. Ellsworth Rd Ann Arbor, MI USA TABLE OF CONTENTS GENERAL

More information

For detecting mitochondrial membrane potential changes in cells using our proprietary fluorescence probe.

For detecting mitochondrial membrane potential changes in cells using our proprietary fluorescence probe. ab112133 JC-10 Mitochondrial Membrane Potential Assay Kit Flow Cytometry Instructions for Use For detecting mitochondrial membrane potential changes in cells using our proprietary fluorescence probe. This

More information

The measurement of telomere length was performed by the same method as in the previous study (11).

The measurement of telomere length was performed by the same method as in the previous study (11). 1 SUPPLEMENTAL DATA 2 3 METHODS 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 Telomere length measurement by quantitative real-time PCR (q-pcr) The measurement of telomere length was performed

More information

Measuring cell proliferation using the CyQUANT Cell Proliferation Assay with SpectraMax Microplate Readers

Measuring cell proliferation using the CyQUANT Cell Proliferation Assay with SpectraMax Microplate Readers APPLICATION NOTE Measuring cell proliferation using the CyQUANT Cell Proliferation Assay with SpectraMax Microplate Readers Introduction Quantitation of cell proliferation using fluorescence allows one

More information

Characterizing Phenotypes of Bacteria by Staining Method

Characterizing Phenotypes of Bacteria by Staining Method Experiment 3 Laboratory to Biology III Diversity of Microorganisms / Wintersemester / page 1 Experiment 3 Characterizing Phenotypes of Bacteria by Staining Method Advisor NN Reading Chapters in BBOM 9

More information

Mitochondrial Membrane Potential Detection Kit

Mitochondrial Membrane Potential Detection Kit Mitochondrial Membrane Potential Detection Kit Instruction Manual Catalog #280002 Mitochondrial Membrane Potential Detection Kit Revision B Research Use Only. Not for Use in Diagnostic Procedures. 280002-12

More information

Characterizing Phenotypes of Bacteria by Staining Method

Characterizing Phenotypes of Bacteria by Staining Method Experiment 3 Laboratory to Biology III Diversity of Microorganisms / Wintersemester / page 1 Experiment Characterizing Phenotypes of Bacteria by Staining Method Advisor Reading NN Chapters 3.1, 3.7, 3.8,

More information

ab MDR Assay Kit - flow cytometry (Fluorometric - gold)

ab MDR Assay Kit - flow cytometry (Fluorometric - gold) ab204535 MDR Assay Kit - flow cytometry (Fluorometric - gold) Instructions for Use For functional detection and profiling of multidrug resistant phenotypes in live cells (both suspension and adherent).

More information

Complement Titration Tray Class I and II. Instructions for Use

Complement Titration Tray Class I and II. Instructions for Use Complement Titration Tray Class I and II Instructions for Use Complement Titration Tray, Class I and II Instructions for Use The Invitrogen Complement Titration Tray, Class I and II, is a serological based

More information

Electronic Supplementary Material (ESI) for Journal of Materials Chemistry This journal is The Royal Society of Chemistry 2011.

Electronic Supplementary Material (ESI) for Journal of Materials Chemistry This journal is The Royal Society of Chemistry 2011. Experimental: MTT assay: To determine cell viability the colorimetric MTT metabolic activity assay was used. Hela cells (1 10 4 cells/well) were cultured in a 96-well plate at 37 C, and exposed to varying

More information

The Basics of Flow Cytometry

The Basics of Flow Cytometry The Basics of Flow Cytometry F ACS C ore F acility Janine Bögli, Biozentrum, 29. January 2018 The functions of the FACS Core Facility Centralization of equipment and expertise Train users Sorter operation

More information

Your Research, Revolutionized

Your Research, Revolutionized Your Research, Revolutionized Drive Your Research Forward Your research needs are evolving and with the CytoFLEX flow cytometer you ll see just how far your data can take you. CytoFLEX has the advanced

More information

Living and Dead Cells Staining: -Cellstain- Double Staining Kit

Living and Dead Cells Staining: -Cellstain- Double Staining Kit Introduction -Cellstain - Double Staining Kit combines Calcein-AM (used for fluorescent staining the living cells) and Propidium Iodide (used for a fluorescent staining of the dead cells) for simultaneous

More information

ab Nitric Oxide Assay Kit (Flow cytometry -

ab Nitric Oxide Assay Kit (Flow cytometry - Version 1 Last updated 25 April 2017 ab219933 Nitric Oxide Assay Kit (Flow cytometry - Orange) For the rapid, sensitive and accurate measurement of NO in live cells This product is for research use only

More information

FACSCalibur. FACS200 Handbook Pro DNA. FACSCalibur FACSCalibur. FACS Calibur. FSC Diode 488 nm SSC PMT 488 nm. FACS200 Handbook Pro 1

FACSCalibur. FACS200 Handbook Pro DNA. FACSCalibur FACSCalibur. FACS Calibur. FSC Diode 488 nm SSC PMT 488 nm. FACS200 Handbook Pro 1 FACSCalibur FACS200 Handbook Pro DNA FACSCalibur 1.1 1. FACSCalibur 2. FACS Calibur 488 nm FACS Calibur FSC Diode 488 nm SSC PMT 488 nm FL1 PMT 515-545 nm FL2 PMT 564-606 nm FL3 PMT >650 nm FACS200 Handbook

More information

The Cell Cycle, DNA Replication, and Mitosis

The Cell Cycle, DNA Replication, and Mitosis Chapter 19 The Cell Cycle, DNA Replication, and Mitosis 首頁 OUTLINE Overview of the Cell Cycle DNA Replication DNA Damage and Repair Nuclear and Cell Division Regulation of the Cell Cycle Growth Factors

More information

CytoPainter Cell Proliferation Staining Reagent Blue Fluorescence (Ex-405nm)

CytoPainter Cell Proliferation Staining Reagent Blue Fluorescence (Ex-405nm) ab176726 CytoPainter Cell Proliferation Staining Reagent Blue Fluorescence (Ex-405nm) Instructions for Use For fluorescent labeling and analysis on heterogeneous cell populations. This product is for research

More information

Flexible, Intuitive and Affordable

Flexible, Intuitive and Affordable Data Sheet guava easycyte Flow Cytometry Systems Flexible, Intuitive and Affordable The guava flow cytometry systems are easy to use and deliver complete and comprehensive cell analysis right on your benchtop.

More information

Before You Begin. Calibration Protocols

Before You Begin. Calibration Protocols Before You Begin Read through this entire protocol sheet carefully before you start your experiment and prepare any materials you may need. Calibration Protocols 1. OD 600 Reference point You will use

More information

LacZ beta Galactosidase Intracellular Detection Kit

LacZ beta Galactosidase Intracellular Detection Kit ab189816 LacZ beta Galactosidase Intracellular Detection Kit Instructions for Use For the detection of beta-galactosidase using Microplate or FACS Assay This product is for research use only and is not

More information

Flow cytometry within reach.

Flow cytometry within reach. Affordable, but more than capable. The Accuri C6 Flow Cytometer is flow cytometry at its best. Its affordable price makes it accessible beyond compare. The C6 has a foot print small enough to fit most

More information

Comparison of flow cytometry and spectrofluorometric measurements in cell permeabilization experiments

Comparison of flow cytometry and spectrofluorometric measurements in cell permeabilization experiments Comparison of flow cytometry and spectrofluorometric measurements in cell permeabilization experiments L10 Janja Dermol, Lea Vukanović University of Ljubljana, Faculty of Electrical Engineering Duration

More information

APO-BrdU TUNEL Assay Kit

APO-BrdU TUNEL Assay Kit USER GUIDE APO-BrdU TUNEL Assay Kit Catalog No. A23210 Pub. No. MAN0002269 Rev. A.0 Table 1. Contents and storage Material Amount Storage* Stability A35125 APO-BrdU TUNEL Assay Kit 20 C Components Positive

More information

Navios EX FLOW CYTOMETER POWERFUL, DEPENDABLE FLOW CYTOMETRY

Navios EX FLOW CYTOMETER POWERFUL, DEPENDABLE FLOW CYTOMETRY Navios EX FLOW CYTOMETER POWERFUL, DEPENDABLE FLOW CYTOMETRY BECAUSE EVERY EVENT MATTERS The Navios EX flow cytometer offers a solution for advanced cytometry applications with optimized workflows for

More information

Minimum Information about a Flow Cytometry Experiment (MIFlowCyt) Annotation

Minimum Information about a Flow Cytometry Experiment (MIFlowCyt) Annotation Minimum Information about a Flow Cytometry Experiment (MIFlowCyt) Annotation 1. Experiment Overview 1.1 Purpose The purpose of these sets of experiments is to develop a methodology of identifying and quantifying

More information

Application Note. Assay Portability on the BD FACSVerse System. Summary. Maria Jaimes, Yibing Wang, Catherine McIntyre, and Dev Mittar

Application Note. Assay Portability on the BD FACSVerse System. Summary. Maria Jaimes, Yibing Wang, Catherine McIntyre, and Dev Mittar September Assay Portability on the BD FACSVerse System Maria Jaimes, Yibing Wang, Catherine McIntyre, and Dev Mittar Contents Summary Introduction 3 Objective 4 Methods 6 Results Discussion Conclusions

More information

JC 1 Mitochondrial Membrane Potential Assay

JC 1 Mitochondrial Membrane Potential Assay G-Biosciences 1-800-628-7730 1-314-991-6034 technical@gbiosciences.com A Geno Technology, Inc. (USA) brand name JC 1 Mitochondrial Membrane Potential Assay (Cat. # 786 1321, 786 1322) think proteins! think

More information

E. coli Phagocytosis Assay Kit

E. coli Phagocytosis Assay Kit E. coli Phagocytosis Assay Kit Item No. 601370 www.caymanchem.com Customer Service 800.364.9897 Technical Support 888.526.5351 1180 E. Ellsworth Rd Ann Arbor, MI USA TABLE OF CONTENTS GENERAL INFORMATION

More information

ab Nitric Oxide Assay Kit (Flow cytometry - Red)

ab Nitric Oxide Assay Kit (Flow cytometry - Red) Version 1 Last updated 26 October 2018 ab219934 Nitric Oxide Assay Kit (Flow cytometry - Red) For the rapid, sensitive and accurate measurement of NO in live cells This product is for research use only

More information

Boundary-breaking acoustic focusing cytometry

Boundary-breaking acoustic focusing cytometry Boundary-breaking acoustic focusing cytometry Introducing the Attune NxT Acoustic Focusing Cytometer a high-performance system that s flexible enough for any lab One of the main projects in my laboratory

More information

CFSE Cell Division Assay Kit

CFSE Cell Division Assay Kit CFSE Cell Division Assay Kit Item No. 10009853 www.caymanchem.com Customer Service 800.364.9897 Technical Support 888.526.5351 1180 E. Ellsworth Rd Ann Arbor, MI USA TABLE OF CONTENTS GENERAL INFORMATION

More information

ab Nitric Oxide Assay Kit (Flow cytometry - Red)

ab Nitric Oxide Assay Kit (Flow cytometry - Red) Version 1 Last updated 25 April 2017 ab219934 Nitric Oxide Assay Kit (Flow cytometry - Red) For the rapid, sensitive and accurate measurement of NO in live cells This product is for research use only and

More information

Comparing Fluorescence-Based Viability Detection Method using the Cellometer Vision

Comparing Fluorescence-Based Viability Detection Method using the Cellometer Vision Comparing Fluorescence-Based Viability Detection Method using the Cellometer Vision Nexcelom Bioscience LLC. 360 Merrimack Street, Building 9 Lawrence, MA 01843 T: 978.327.5340 F: 978.327.5341 E: info@nexcelom.com

More information

Flow Cytometry Immune Activation SOP

Flow Cytometry Immune Activation SOP Flow Cytometry Immune Activation SOP Purpose This SOP standardizes the procedure for measuring immune activation of T cells using flow cytometry in ACTG Immunology Laboratories. Materials 1. 12x75mm flow

More information

Designing and Implementing a High-Level Multicolor Flow Cytometry Assay. Brent Wood MD PhD Department of Laboratory Medicine University of Washington

Designing and Implementing a High-Level Multicolor Flow Cytometry Assay. Brent Wood MD PhD Department of Laboratory Medicine University of Washington Designing and Implementing a High-Level Multicolor Flow Cytometry Assay Brent Wood MD PhD Department of Laboratory Medicine University of Washington Define Purpose of Assay Most important question What

More information