FITC, fluorescein isothiocyanate. 772 , 312 (2011). Acridine orange is an organic compound that serves as a nucleic acid-selective fluorescent dye with cationic properties useful for cell cycle determination. The LIVE/DEAD BacLight Bacterial Viability and Counting Kit allows researchers to reliably distinguish and quantitate live and dead bacteria with the aid of a flow cytometer, even in a mixed population containing a range of bacterial types. Molecular Probes fluorescence reagents are among the most peer-referenced in all of life science research. It is critical to understand the degree of cell death in any flow cytometry assay and exclude those cells from the analysis. J Microbiol Methods. The biocompatibility of the gels was evaluated by fluorescein diacetate (FDA)/propidium iodide (PI) staining, immunofluorescence staining of biomarkers, and Cell Counting Kit-8 (CCK8) assay of human umbilical vein endothelial cells (HUVECs) and vascular smooth muscle cells (VSMCs), which are the main cell components of blood vessels. Using the N/TERT-1 immortalized human keratinocyte cell line (hereafter referred to as N/TERTs), we confirmed published findings (15, 27) that UVB irradiation causes NLRP1-dependent pyroptosis in a dose-dependent manner as measured by IL-1 secretion, GSDMD cleavage, ASC oligomerization, and rapid propidium iodide (PI) uptake (within 4 to 6 hours) (fig. Pathogenic drug-resistant bacteria represent a threat to human health, for instance, the methicillin-resistant Staphylococcus aureus (MRSA). both the cytoplasm and nuclei of eukaryotic cells show staining with hexidium iodide; however, mitochondria and nucleoli may also be stained. Cell cycle profiles were measured using flow cytometry based on the DNA content of cells using propidium iodide (Sigma-Aldrich, P4170) staining as described previously 62. The permeability of bacteria membrane was further examined by propidium iodide (PI) staining, and Hoechst 33342 was used to label bacteria. Staining is a technique used to enhance contrast in samples, generally at the microscopic level. For propidium iodide (PI) staining and GFP observation, Arabidopsis seedlings of TIE1pro-TIE1-GFP or TIE2pro-TIE2-GFP were grown vertically on 1/2 MS medium. It is critical to understand the degree of cell death in any flow cytometry assay and exclude those cells from the analysis. Bacteria can bypass the blood-brain barrier (BBB), suggesting the possibility of employment of bacteria for combating central nervous system diseases. It is critical to understand the degree of cell death in any flow cytometry assay and exclude those cells from the analysis. BioLegend provides DNA dyes, Propidium Iodide and 7- AAD, that enter and stain dead cells, but are impermeable to live cells for rapid, cost- effective analysis of unfixed cells. 2016;131:17280. 2016;131:17280. For reconstitution, pre-warm the kit to room temperature; add 100 l of DMSO to one vial of Zombie Aqua dye and mix until fully dissolved. i,j, BMDM apoptosis triggered by LPS (100 ng ml 1, 48 h) analysed by annexin V and propidium iodide double staining. FITC, fluorescein isothiocyanate. Cells with a compromised membrane that are considered to be dead or dying will stain red, whereas cells with an intact membrane will stain green. Methods Mol. Cell viability assays are used to measure the physical and physiological health of cells in response to extracellular stimuli, chemical agents, or therapeutic treatments [13], or when determining optimal growth conditions in In cases where cell fixation is required, we now introduce fixable Zombie Aqua. The LIVE/DEAD BacLight Bacterial Viability Kit *for microscopy and quantitative assays* is a convenient and easy-to-use kit for monitoring the viability of bacterial populations as a function of the membrane integrity of the cell. Other nuclear stains include Hoechst 33342, which is cell permanent and can be used with live as well as fixed cells, and propidium iodide, long used as a nuclear marker for flow cytometry which fluoresces in the red range. Stains and dyes are frequently used in histology (microscopic study of biological tissues), in cytology (microscopic study of cells), and in the medical fields of histopathology, hematology, and cytopathology that focus on the study and diagnoses of diseases at the microscopic level. Data subsequently were analysed with FlowJo software. The RSG dye can easily penetrate bacteria and yield green fluorescence when reduced by (Propidium iodide) staining. The RSG dye can easily penetrate bacteria and yield green fluorescence when reduced by (Propidium iodide) staining. J Microbiol Methods. Flow cytometry (FACS) staining protocol (Cell surface staining) Harvest, wash the cells (single cell suspension) and adjust cell number to a concentration of 1-5x106 cells/ml in ice cold FACS Buffer (PBS, 0.5-1% BSA or 5-10% FBS, 0.1% NaN3 sodium azide*). 772 , 312 (2011). BioLegend provides DNA dyes, Propidium Iodide and 7- AAD, that enter and stain dead cells, but are impermeable to live cells for rapid, cost- effective analysis of unfixed cells. Biol. A pipeline for developing and testing staining protocols for flow cytometry, demonstrated with SYBR Green I and propidium iodide viability staining. Acridine orange is cell-permeable, which allows the dye to interact with DNA by intercalation, or RNA via electrostatic attractions.When bound to DNA, acridine orange is very similar spectrally to an organic Resuspend 1-10 x 10 6 cells in the appropriate volume of Zombie NIR solution. Molecular Probes fluorescence reagents are among the most peer-referenced in all of life science research. (D) Live/dead cell staining fluorescence images of CT26 cells stained by calcein AM/propidium iodide (PI) after incubation with different nanoplatforms for 24 hours (scale bars, 100 m). Flow cytometry (FACS) staining protocol (Cell surface staining) Harvest, wash the cells (single cell suspension) and adjust cell number to a concentration of 1-5x106 cells/ml in ice cold FACS Buffer (PBS, 0.5-1% BSA or 5-10% FBS, 0.1% NaN3 sodium azide*). Biol. Staining is a technique used to enhance contrast in samples, generally at the microscopic level. PI can penetrate the destroyed cell Acridine orange is cell-permeable, which allows the dye to interact with DNA by intercalation, or RNA via electrostatic attractions.When bound to DNA, acridine orange is very similar spectrally to an organic Resuspend 1-10 x 10 6 cells in the appropriate volume of Zombie NIR solution. Other nuclear stains include Hoechst 33342, which is cell permanent and can be used with live as well as fixed cells, and propidium iodide, long used as a nuclear marker for flow cytometry which fluoresces in the red range. Herein, we first reported the diameter- and shape-controlled synthesis of Mg nanoparticles, including hexagonal nanosheet (80320 nm), nanoflower (100250 nm), and small After fixation at 4 C for at least 16 h, cells were stained with propidium iodide, and at least 10,000 cells were analysed by FACS. It is especially useful for staining both gram-positive and gram-negative bacteria, in which an exceptionally bright signal is required. Using the N/TERT-1 immortalized human keratinocyte cell line (hereafter referred to as N/TERTs), we confirmed published findings (15, 27) that UVB irradiation causes NLRP1-dependent pyroptosis in a dose-dependent manner as measured by IL-1 secretion, GSDMD cleavage, ASC oligomerization, and rapid propidium iodide (PI) uptake (within 4 to 6 hours) (fig. Its selectivity for DNA allows efficient staining of nuclei with little background from the cytoplasm. Bacteria were collected and sonicated in the lysis buffer containing 20 mM Tris-HCl (pH 8.0), 150 mM NaCl, 20 mM imidazole and 10 mM 2-mercaptoethanol. After fixation at 4 C for at least 16 h, cells were stained with propidium iodide, and at least 10,000 cells were analysed by FACS. Different cell types can have a wide degree of variability in staining based on cell size and degree of cell death. After the antibody reaction, cells were washed by FACS buffer and re-suspend to 0.1% Propidium iodide (PI) containing FACS buffer. Cell viability refers to the number of live, healthy cells in a sample [1]. The LIVE/DEAD BacLight Bacterial Viability and Counting Kit allows researchers to reliably distinguish and quantitate live and dead bacteria with the aid of a flow cytometer, even in a mixed population containing a range of bacterial types. Therefore, propidium iodide (PI), a fluorescent and intercalating agent, is widely used as DNA stain . Cell viability assays are used to measure the physical and physiological health of cells in response to extracellular stimuli, chemical agents, or therapeutic treatments [13], or when determining optimal growth conditions in Roots were stained with PI solution (1 mg/ml; Sigma-Aldrich, P4170) for 30 s. PI staining and GFP were then observed using a confocal laser scanning microscope (Zeiss LSM 710 NLO). Panel A shows the dot plot of forward scatter vs. side scatter of an untreated Saccharomyces culture, washed and stained with SYTO 9 dye and propidium iodide as described in the LIVE/DEAD FungaLight Yeast Viability Kit protocol. For reconstitution, pre-warm the kit to room temperature; add 100 l of DMSO to one vial of Zombie Aqua dye and mix until fully dissolved. In another experiment, Panc02 cells were incubated with DMEM containing P(C6-Bn 20)-cy5 (40 g ml 1), propidium iodide (10 g ml 1) and Annexin V-FITC (5 l) at pH 6.8. S1A). After the antibody reaction, cells were washed by FACS buffer and re-suspend to 0.1% Propidium iodide (PI) containing FACS buffer. After fixation at 4 C for at least 16 h, cells were stained with propidium iodide, and at least 10,000 cells were analysed by FACS. Pathogenic drug-resistant bacteria represent a threat to human health, for instance, the methicillin-resistant Staphylococcus aureus (MRSA). The purpose of present protocol is to assess the toxigenic potential of selected/isolated probiotic organism using MTT assay. Flow cytometry (FACS) staining protocol (Cell surface staining) Harvest, wash the cells (single cell suspension) and adjust cell number to a concentration of 1-5x106 cells/ml in ice cold FACS Buffer (PBS, 0.5-1% BSA or 5-10% FBS, 0.1% NaN3 sodium azide*). Resuspend 1-10 x 10 6 cells in the appropriate volume of Zombie NIR solution. To quantitatively measure mitochondrial fragmentation in WT and Lrrk2 G2019S BMDMs, we developed a strategy to measure mitochondrial size differences down to the sub-micron level via flow cytometry. CAS PubMed Article Google Scholar Cells with a compromised membrane that are considered to be dead or dying will stain red, whereas cells with an intact membrane will stain green. The purpose of present protocol is to assess the toxigenic potential of selected/isolated probiotic organism using MTT assay. BioLegend provides DNA dyes, Propidium Iodide and 7- AAD, that enter and stain dead cells, but are impermeable to live cells for rapid, cost- effective analysis of unfixed cells. Bacteria reportedly produce more EPS under stress conditions, was prepared according to a previously published protocol (Tantra et al., after the injection of 3 mL of 3.34 mM Syto9 and 3 mL of 4.67 mM propidium iodide (Molecular Probes, Carlsbad, CA) into each flow channel to stain live and dead cells in green and red, respectively. (D) Live/dead cell staining fluorescence images of CT26 cells stained by calcein AM/propidium iodide (PI) after incubation with different nanoplatforms for 24 hours (scale bars, 100 m). In another experiment, Panc02 cells were incubated with DMEM containing P(C6-Bn 20)-cy5 (40 g ml 1), propidium iodide (10 g ml 1) and Annexin V-FITC (5 l) at pH 6.8. Methods Mol. Various bacteria have been isolated from different sources and believed to be safe for using them as probiotics. Data subsequently were analysed with FlowJo software. Hare, E. E. & Johnston, J. S. Genome size determination using flow cytometry of propidium iodide-stained nuclei. Bacteria were collected and sonicated in the lysis buffer containing 20 mM Tris-HCl (pH 8.0), 150 mM NaCl, 20 mM imidazole and 10 mM 2-mercaptoethanol. (D) Live/dead cell staining fluorescence images of CT26 cells stained by calcein AM/propidium iodide (PI) after incubation with different nanoplatforms for 24 hours (scale bars, 100 m). The Gram-negative bacteria Legionella pneumophila triggers multiple inflammasomes and is as a reliable model to infected with L. pneumophila occurs quickly after infection and can be easily assessed by probing the influx of propidium iodide (PI) to the cell nuclei (fig. In cases where cell fixation is required, we now introduce fixable Zombie Aqua. The biocompatibility of the gels was evaluated by fluorescein diacetate (FDA)/propidium iodide (PI) staining, immunofluorescence staining of biomarkers, and Cell Counting Kit-8 (CCK8) assay of human umbilical vein endothelial cells (HUVECs) and vascular smooth muscle cells (VSMCs), which are the main cell components of blood vessels. For propidium iodide (PI) staining and GFP observation, Arabidopsis seedlings of TIE1pro-TIE1-GFP or TIE2pro-TIE2-GFP were grown vertically on 1/2 MS medium. The RSG dye can easily penetrate bacteria and yield green fluorescence when reduced by (Propidium iodide) staining. 100 tests = 1 vial of Zombie Aqua + DMSO, 500 tests = 5 vials of Zombie Aqua + DMSO. FITC, fluorescein isothiocyanate. Cells with a compromised membrane that are considered to be dead or dying will stain red, whereas cells with an intact membrane will stain green. S1A). FITC, fluorescein isothiocyanate. It is especially useful for staining both gram-positive and gram-negative bacteria, in which an exceptionally bright signal is required. i,j, BMDM apoptosis triggered by LPS (100 ng ml 1, 48 h) analysed by annexin V and propidium iodide double staining. 100 tests = 1 vial of Zombie Aqua + DMSO, 500 tests = 5 vials of Zombie Aqua + DMSO. For example, if you are adding 20 l of antibody cocktail for a 100 l total staining volume, use 80 l of Zombie NIR solution. EdU assay Stains and dyes are frequently used in histology (microscopic study of biological tissues), in cytology (microscopic study of cells), and in the medical fields of histopathology, hematology, and cytopathology that focus on the study and diagnoses of diseases at the microscopic level. Bacteria can bypass the blood-brain barrier (BBB), suggesting the possibility of employment of bacteria for combating central nervous system diseases. Molecular Probes fluorescence reagents are among the most peer-referenced in all of life science research. Pathogenic drug-resistant bacteria represent a threat to human health, for instance, the methicillin-resistant Staphylococcus aureus (MRSA). However, in vitro toxicity must be assessed as some bacteria may be toxic to the host. Cell viability refers to the number of live, healthy cells in a sample [1]. However, in vitro toxicity must be assessed as some bacteria may be toxic to the host. For propidium iodide (PI) staining and GFP observation, Arabidopsis seedlings of TIE1pro-TIE1-GFP or TIE2pro-TIE2-GFP were grown vertically on 1/2 MS medium. Cell cycle profiles were measured using flow cytometry based on the DNA content of cells using propidium iodide (Sigma-Aldrich, P4170) staining as described previously 62. Its selectivity for DNA allows efficient staining of nuclei with little background from the cytoplasm. However, in vitro toxicity must be assessed as some bacteria may be toxic to the host. Its selectivity for DNA allows efficient staining of nuclei with little background from the cytoplasm. It is especially useful for staining both gram-positive and gram-negative bacteria, in which an exceptionally bright signal is required. Trusted product bands including Alexa Fluor, SYTOX and Click-iT empower scientists working in cell biology, genetic analysis, protein biology, and cell engineering to make the discoveries that advance our understanding today and catalyze the research goals of tomorrow. J Microbiol Methods. Other nuclear stains include Hoechst 33342, which is cell permanent and can be used with live as well as fixed cells, and propidium iodide, long used as a nuclear marker for flow cytometry which fluoresces in the red range. 100 tests = 1 vial of Zombie Aqua + DMSO, 500 tests = 5 vials of Zombie Aqua + DMSO. In another experiment, Panc02 cells were incubated with DMEM containing P(C6-Bn 20)-cy5 (40 g ml 1), propidium iodide (10 g ml 1) and Annexin V-FITC (5 l) at pH 6.8. Product Details Preparation Zombie Aqua Fixable Viability Kit is composed of lyophilized Zombie Aqua dye and anhydrous DMSO. To quantitatively measure mitochondrial fragmentation in WT and Lrrk2 G2019S BMDMs, we developed a strategy to measure mitochondrial size differences down to the sub-micron level via flow cytometry. The Gram-negative bacteria Legionella pneumophila triggers multiple inflammasomes and is as a reliable model to infected with L. pneumophila occurs quickly after infection and can be easily assessed by probing the influx of propidium iodide (PI) to the cell nuclei (fig. In cases where cell fixation is required, we now introduce fixable Zombie Aqua. The purpose of present protocol is to assess the toxigenic potential of selected/isolated probiotic organism using MTT assay. The permeability of bacteria membrane was further examined by propidium iodide (PI) staining, and Hoechst 33342 was used to label bacteria. PI can penetrate the destroyed cell Then, the antibacterial mechanism was studied. Cell cycle profiles were measured using flow cytometry based on the DNA content of cells using propidium iodide (Sigma-Aldrich, P4170) staining as described previously 62. To quantitatively measure mitochondrial fragmentation in WT and Lrrk2 G2019S BMDMs, we developed a strategy to measure mitochondrial size differences down to the sub-micron level via flow cytometry. Hare, E. E. & Johnston, J. S. Genome size determination using flow cytometry of propidium iodide-stained nuclei. Trusted product bands including Alexa Fluor, SYTOX and Click-iT empower scientists working in cell biology, genetic analysis, protein biology, and cell engineering to make the discoveries that advance our understanding today and catalyze the research goals of tomorrow. The biocompatibility of the gels was evaluated by fluorescein diacetate (FDA)/propidium iodide (PI) staining, immunofluorescence staining of biomarkers, and Cell Counting Kit-8 (CCK8) assay of human umbilical vein endothelial cells (HUVECs) and vascular smooth muscle cells (VSMCs), which are the main cell components of blood vessels. For example, if you are adding 20 l of antibody cocktail for a 100 l total staining volume, use 80 l of Zombie NIR solution. CAS PubMed Article Google Scholar Hare, E. E. & Johnston, J. S. Genome size determination using flow cytometry of propidium iodide-stained nuclei. Metallic magnesium is a promising platform for H 2 cancer therapy, whereas the available Mg particles undesirable diameter and shape greatly hinder its therapeutic application. Then, the antibacterial mechanism was studied. Polystyrene beads of 1, 2, and 4 m were used to create a gating strategy, whereby the size of MitoTracker green (MTG)-stained mitochondria could be EdU assay Various bacteria have been isolated from different sources and believed to be safe for using them as probiotics. Staining is a technique used to enhance contrast in samples, generally at the microscopic level. Bacteria were collected and sonicated in the lysis buffer containing 20 mM Tris-HCl (pH 8.0), 150 mM NaCl, 20 mM imidazole and 10 mM 2-mercaptoethanol. Roots were stained with PI solution (1 mg/ml; Sigma-Aldrich, P4170) for 30 s. PI staining and GFP were then observed using a confocal laser scanning microscope (Zeiss LSM 710 NLO). Bacteria can bypass the blood-brain barrier (BBB), suggesting the possibility of employment of bacteria for combating central nervous system diseases. Metallic magnesium is a promising platform for H 2 cancer therapy, whereas the available Mg particles undesirable diameter and shape greatly hinder its therapeutic application. Different cell types can have a wide degree of variability in staining based on cell size and degree of cell death. Different cell types can have a wide degree of variability in staining based on cell size and degree of cell death. Roots were stained with PI solution (1 mg/ml; Sigma-Aldrich, P4170) for 30 s. PI staining and GFP were then observed using a confocal laser scanning microscope (Zeiss LSM 710 NLO). PI can penetrate the destroyed cell 2016;131:17280. Trusted product bands including Alexa Fluor, SYTOX and Click-iT empower scientists working in cell biology, genetic analysis, protein biology, and cell engineering to make the discoveries that advance our understanding today and catalyze the research goals of tomorrow. Panel A shows the dot plot of forward scatter vs. side scatter of an untreated Saccharomyces culture, washed and stained with SYTO 9 dye and propidium iodide as described in the LIVE/DEAD FungaLight Yeast Viability Kit protocol. Then, the antibacterial mechanism was studied. A pipeline for developing and testing staining protocols for flow cytometry, demonstrated with SYBR Green I and propidium iodide viability staining. Polystyrene beads of 1, 2, and 4 m were used to create a gating strategy, whereby the size of MitoTracker green (MTG)-stained mitochondria could be Product Details Preparation Zombie Aqua Fixable Viability Kit is composed of lyophilized Zombie Aqua dye and anhydrous DMSO. both the cytoplasm and nuclei of eukaryotic cells show staining with hexidium iodide; however, mitochondria and nucleoli may also be stained. Acridine orange is an organic compound that serves as a nucleic acid-selective fluorescent dye with cationic properties useful for cell cycle determination. EdU assay The LIVE/DEAD BacLight Bacterial Viability Kit *for microscopy and quantitative assays* is a convenient and easy-to-use kit for monitoring the viability of bacterial populations as a function of the membrane integrity of the cell. Cell viability refers to the number of live, healthy cells in a sample [1]. Various bacteria have been isolated from different sources and believed to be safe for using them as probiotics. 772 , 312 (2011). The Gram-negative bacteria Legionella pneumophila triggers multiple inflammasomes and is as a reliable model to infected with L. pneumophila occurs quickly after infection and can be easily assessed by probing the influx of propidium iodide (PI) to the cell nuclei (fig. Acridine orange is an organic compound that serves as a nucleic acid-selective fluorescent dye with cationic properties useful for cell cycle determination. Biol. Using the N/TERT-1 immortalized human keratinocyte cell line (hereafter referred to as N/TERTs), we confirmed published findings (15, 27) that UVB irradiation causes NLRP1-dependent pyroptosis in a dose-dependent manner as measured by IL-1 secretion, GSDMD cleavage, ASC oligomerization, and rapid propidium iodide (PI) uptake (within 4 to 6 hours) (fig. The LIVE/DEAD BacLight Bacterial Viability and Counting Kit allows researchers to reliably distinguish and quantitate live and dead bacteria with the aid of a flow cytometer, even in a mixed population containing a range of bacterial types. FITC, fluorescein isothiocyanate. Stains and dyes are frequently used in histology (microscopic study of biological tissues), in cytology (microscopic study of cells), and in the medical fields of histopathology, hematology, and cytopathology that focus on the study and diagnoses of diseases at the microscopic level. Polystyrene beads of 1, 2, and 4 m were used to create a gating strategy, whereby the size of MitoTracker green (MTG)-stained mitochondria could be Panel A shows the dot plot of forward scatter vs. side scatter of an untreated Saccharomyces culture, washed and stained with SYTO 9 dye and propidium iodide as described in the LIVE/DEAD FungaLight Yeast Viability Kit protocol. S1A). FITC, fluorescein isothiocyanate. Metallic magnesium is a promising platform for H 2 cancer therapy, whereas the available Mg particles undesirable diameter and shape greatly hinder its therapeutic application. Bacteria reportedly produce more EPS under stress conditions, was prepared according to a previously published protocol (Tantra et al., after the injection of 3 mL of 3.34 mM Syto9 and 3 mL of 4.67 mM propidium iodide (Molecular Probes, Carlsbad, CA) into each flow channel to stain live and dead cells in green and red, respectively. i,j, BMDM apoptosis triggered by LPS (100 ng ml 1, 48 h) analysed by annexin V and propidium iodide double staining. Methods Mol. A pipeline for developing and testing staining protocols for flow cytometry, demonstrated with SYBR Green I and propidium iodide viability staining. For example, if you are adding 20 l of antibody cocktail for a 100 l total staining volume, use 80 l of Zombie NIR solution. After the antibody reaction, cells were washed by FACS buffer and re-suspend to 0.1% Propidium iodide (PI) containing FACS buffer. Therefore, propidium iodide (PI), a fluorescent and intercalating agent, is widely used as DNA stain . CAS PubMed Article Google Scholar Herein, we first reported the diameter- and shape-controlled synthesis of Mg nanoparticles, including hexagonal nanosheet (80320 nm), nanoflower (100250 nm), and small For reconstitution, pre-warm the kit to room temperature; add 100 l of DMSO to one vial of Zombie Aqua dye and mix until fully dissolved. The permeability of bacteria membrane was further examined by propidium iodide (PI) staining, and Hoechst 33342 was used to label bacteria. Cell viability assays are used to measure the physical and physiological health of cells in response to extracellular stimuli, chemical agents, or therapeutic treatments [13], or when determining optimal growth conditions in Therefore, propidium iodide (PI), a fluorescent and intercalating agent, is widely used as DNA stain . Data subsequently were analysed with FlowJo software. Herein, we first reported the diameter- and shape-controlled synthesis of Mg nanoparticles, including hexagonal nanosheet (80320 nm), nanoflower (100250 nm), and small The LIVE/DEAD BacLight Bacterial Viability Kit *for microscopy and quantitative assays* is a convenient and easy-to-use kit for monitoring the viability of bacterial populations as a function of the membrane integrity of the cell. Acridine orange is cell-permeable, which allows the dye to interact with DNA by intercalation, or RNA via electrostatic attractions.When bound to DNA, acridine orange is very similar spectrally to an organic Product Details Preparation Zombie Aqua Fixable Viability Kit is composed of lyophilized Zombie Aqua dye and anhydrous DMSO. both the cytoplasm and nuclei of eukaryotic cells show staining with hexidium iodide; however, mitochondria and nucleoli may also be stained. Bacteria reportedly produce more EPS under stress conditions, was prepared according to a previously published protocol (Tantra et al., after the injection of 3 mL of 3.34 mM Syto9 and 3 mL of 4.67 mM propidium iodide (Molecular Probes, Carlsbad, CA) into each flow channel to stain live and dead cells in green and red, respectively.