propidium iodide staining protocol microscopytracksmith boston singlet 2022

Propidium iodide. Detection of dead cells is accomplished by measuring movement of molecules either into or out of cells 10 5 cells were used for analysis. Resuspended cells were incubated with Annexin V-FITC for 15 min in the dark. Propidium iodide is a fluorescent intercalating agent that can be used to stain cells. Distinct brain regions differentially and rapidly tailor the leukocyte landscape during psychological stress, calibrating the ability of the immune system to respond to physical threats. Apoptotic DNA fragmentation is a key feature of apoptosis, a type of programmed cell death.Apoptosis is characterized by the activation of endogenous endonucleases, particularly the caspase-3 activated DNase (CAD), with subsequent cleavage of nuclear DNA into internucleosomal fragments of roughly 180 base pairs (bp) and multiples thereof (360, 540 etc. No. 556463). 554060) in Propidium Iodide (PI) Staining Solution (Cat. Observe cell staining with fluorescence microscopy . Thus, using Fixable Viability Dyes allows dead cells to be excluded from analysis when intracellular targets are being studied. Blue, MIT; red, cell nuclei stained with propidium iodide (PI). Annexin V and propidium iodide (PI) staining confirmed that NK-92MI cells induced more extensive lytic death in GSDMB-expressing 293T cells than that in cells expressing other gasdermins or the empty vector (fig. PI is widely used in fluorescence microscopy, confocal laser scanning microscopy, flow cytometry, and fluorometry. Apoptotic DNA fragmentation is a key feature of apoptosis, a type of programmed cell death.Apoptosis is characterized by the activation of endogenous endonucleases, particularly the caspase-3 activated DNase (CAD), with subsequent cleavage of nuclear DNA into internucleosomal fragments of roughly 180 base pairs (bp) and multiples thereof (360, 540 etc. Thus, using Fixable Viability Dyes allows dead cells to be excluded from analysis when intracellular targets are being studied. Jurkat cells (Human T-cell leukemia; ATCC TIB-152) were left untreated (top panels) or treated for 4 hours with 12 M campotothecin (bottom panels). For reconstitution, pre-warm the kit to room temperature; add 100 l of DMSO to 3.1 Place the cell suspension from Stage 1.5 on a glass slide. The Invitrogen Molecular Probes Alexa Fluor 488 dyewith spectral properties and quantum yield nearly identical to those of fluorescein isothiocyanate (FITC)produces brighter, more photostable conjugates.These conjugates are ideal for imaging and other applications requiring increased sensitivity and environmentally insensitive TO-PRO-3 and TOTO-3 stains both have much greater extinction coefficients than DNA-bound propidium iodide. Untreated cells were primarily Annexin V-Biotin and PI negative, indicating that they were viable and not undergoing apoptosis. The original propidium iodide staining dataset was resampled to 10 m 3. Add properly diluted primary antibody to cover the sample This stain is commonly used in combination with 5-bromo-2'-deoxyuridine (BrdU) labeling to distinguish the compact chromatin of apoptotic nuclei, to identify replicating cells and to sort cells based on their DNA content. The qPCR protocol was performed for 40 cycles and each cycle consisted of denaturation at 95 C for 15 s, annealing at 60 C for 1 min, and extension at 72 C for 1 min. Untreated cells were primarily Annexin V-Biotin and PI negative, indicating that they were viable and not undergoing apoptosis. 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. Propidium iodide is used as a DNA stain in flow cytometry to evaluate cell viability or DNA content in cell cycle analysis, or in microscopy to visualise Membrane integrity is the feature most often used to detect whether eukaryotic cells cultured in vitro are alive or dead. 1d and Supplementary Fig. j, Propidium iodide staining of tumour-cell pyroptosis induced by treatment with NPGSDMA3 and Phe-BF 3 in mice. 3.1 Place the cell suspension from Stage 1.5 on a glass slide. HUVECs with compromised DNA integrity and/or membranes were indicated by propidium iodide (PI) staining with a 22 h treatment (Fig. Left: negative control - AG6173 untreated cells. Expand Your Options for Multicolor Labeling This stain is commonly used in combination with 5-bromo-2'-deoxyuridine (BrdU) labeling to distinguish the compact chromatin of apoptotic nuclei, to identify replicating cells and to sort cells based on their DNA content. Unlike 7-AAD and propidium iodide, cells labeled with Fixable Viability Dyes can be washed, fixed, permabilized, and stained for intracellular antigens without any loss of staining intensity of the dead cells. a, Confocal laser scanning microscopy fluorescence imaging of glioma cells that were treated with each formulation. TO-PRO-3 and TOTO-3 stains both have much greater extinction coefficients than DNA-bound propidium iodide. Dead Cell Apoptosis Kit with Annexin V Alexa Fluor 488 & Propidium Iodide (PI) FC, FM: 499/521 535/617: V13241: Dead Cell Apoptosis Kit with Annexin V FITC and PI: FC: 494/518 Staining of a mixture of heat-killed and untreated Jurkat cells. Distinct brain regions differentially and rapidly tailor the leukocyte landscape during psychological stress, calibrating the ability of the immune system to respond to physical threats. 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. Jurkat cells were stained according to the protocol in the LIVE/DEAD Violet Viability/Vitality Kit. Thus, using Fixable Viability Dyes allows dead cells to be excluded from analysis when intracellular targets are being studied. Propidium iodide (PI) is a popular red-fluorescent nuclear and chromosome counterstain. Propidium iodide is a fluorescent intercalating agent that can be used to stain cells. Membrane integrity is the feature most often used to detect whether eukaryotic cells cultured in vitro are alive or dead. The original propidium iodide staining dataset was resampled to 10 m 3. 556463). Expand Your Options for Multicolor Labeling Propidium iodide is a popular red-fluorescent nuclear and chromosome counterstain. S1, A to C). Scale bars, 100 m. Our labeling reagents enable researchers to create their own labeled biomolecule for use in immunochemistry, fluorescence in situ hybridization (FISH), cell tracing, receptor labeling, and cytochemistry applications as well as for probing biological structure, function, and interactions.. We also offer our fluorophores conjugated to a variety of antibodies, streptavidin, peptides, Use TO-PRO-3 Stain for 2-Color Viability Staining Simultaneous labeling with a green-fluorescent SYTO dye and cell-impermeant TO-PRO-3 stain is frequently used to assess cell viability. Cells were incubated with FITC Annexin V in a buffer containing propidium iodide (PI) and analyzed by flow cytometry. Use TO-PRO-3 Stain for 2-Color Viability Staining Simultaneous labeling with a green-fluorescent SYTO dye and cell-impermeant TO-PRO-3 stain is frequently used to assess cell viability. Detection of dead cells is accomplished by measuring movement of molecules either into or out of cells 1. Dead Cell Apoptosis Kit with Annexin V Alexa Fluor 488 & Propidium Iodide (PI) Kit Contents: Contains 5 vials of annexin V, Alexa Fluor 488 conjugate (250 L per vial), 1 vial of propidium iodide (PI, 100 L), and 1 bottle of annexin binding buffer (5X solution, 50 mL). Since propidium iodide is not permeant to live cells, it is also commonly used to detect dead cells in a population. 554060) in Propidium Iodide (PI) Staining Solution (Cat. Dead Cell Apoptosis Kit with Annexin V Alexa Fluor 488 & Propidium Iodide (PI) Kit Contents: Contains 5 vials of annexin V, Alexa Fluor 488 conjugate (250 L per vial), 1 vial of propidium iodide (PI, 100 L), and 1 bottle of annexin binding buffer (5X solution, 50 mL). Invitrogen Alexa Fluor 488 dye is a bright, green-fluorescent dye with excitation ideally suited to the 488 nm laser line.For stable signal generation in imaging and flow cytometry, Alexa Fluor 488 dye is pH-insensitive over a wide molar range. Cells that have lost membrane integrity and allow movement of otherwise non-permeable molecules are classified as non-viable or dead. 5a . 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. ). S1, A to C). 1d and Supplementary Fig. Blue, MIT; red, cell nuclei stained with propidium iodide (PI). 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. The gasdermin E protein is shown to act as a tumour suppressor: it is cleaved by caspase 3 and granzyme B and leads to pyroptosis of cancer cells, provoking an immune response to the tumour. Product Type: Apoptosis Detection Kit: Flow Cytometer Laser Lines: 488 Typically, a membrane-impermeable dye like propidium iodide is used to identify dead or dying cells with damaged membranes and a viability dye like calcein-AM used to label live cells. Propidium iodide was used as a counterstain to discriminate necrotic/ dead cells from apoptotic cells. PI is widely used in fluorescence microscopy, confocal laser scanning microscopy, flow cytometry, and fluorometry. 556417) followed by incubation with SAv-FITC (Cat. Invitrogen Alexa Fluor 488 dye is a bright, green-fluorescent dye with excitation ideally suited to the 488 nm laser line.For stable signal generation in imaging and flow cytometry, Alexa Fluor 488 dye is pH-insensitive over a wide molar range. 5a . Scale bars, 100 m. We have protocols for staining using DNA binding dyes with and without antibody staining and a protocol for BrdU staining. Observe cell staining with fluorescence microscopy . An independent experiment from that shown in Fig. Propidium iodide (PI) is a popular red-fluorescent nuclear and chromosome counterstain. Learn more about propidium iodide, and propidium iodide containing products. 3.1 Place the cell suspension from Stage 1.5 on a glass slide. Cells that have lost membrane integrity and allow movement of otherwise non-permeable molecules are classified as non-viable or dead. Invitrogen Alexa Fluor 488 dye is a bright, green-fluorescent dye with excitation ideally suited to the 488 nm laser line.For stable signal generation in imaging and flow cytometry, Alexa Fluor 488 dye is pH-insensitive over a wide molar range. Cells were incubated with Annexin V-Biotin, (Cat. Standard Cell Staining Protocol: Prior to reconstitution, spin down the vial of lyophilized reagent in a microcentrofuge to ensure the reagent is at the bottom of the vial. Probes with high fluorescence quantum yield and high photostability allow detection of low-abundance biological structures with great sensitivity. 2.2 If propidium iodide was added, analyze PI staining by the phycoerythrin emission signal detector (usually FL2). HUVECs with compromised DNA integrity and/or membranes were indicated by propidium iodide (PI) staining with a 22 h treatment (Fig. Propidium iodide is a popular red-fluorescent nuclear and chromosome counterstain. PI is widely used in fluorescence microscopy, confocal laser scanning microscopy, flow cytometry, and fluorometry. 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. Cells that have lost membrane integrity and allow movement of otherwise non-permeable molecules are classified as non-viable or dead. Hoechst 33342 is used for specifically staining the nuclei of living or fixed cells and tissues. Use TO-PRO-3 Stain for 2-Color Viability Staining Simultaneous labeling with a green-fluorescent SYTO dye and cell-impermeant TO-PRO-3 stain is frequently used to assess cell viability. Propidium iodide is a fluorescent intercalating agent that can be used to stain cells. Unlike 7-AAD and propidium iodide, cells labeled with Fixable Viability Dyes can be washed, fixed, permabilized, and stained for intracellular antigens without any loss of staining intensity of the dead cells. For Research Use Only. Dead Cell Apoptosis Kit with Annexin V Alexa Fluor 488 & Propidium Iodide (PI) FC, FM: 499/521 535/617: V13241: Dead Cell Apoptosis Kit with Annexin V FITC and PI: FC: 494/518 Staining of a mixture of heat-killed and untreated Jurkat cells. No. Fluorophore is detected via microscopy; Protocol. An independent experiment from that shown in Fig. Annexin V-FITC/ PI staining of AG06173 primary fibroblasts. Resuspended cells were incubated with Annexin V-FITC for 15 min in the dark. No. 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. 556463). Primary cortical neurons were treated with -syn PFF (5 g/mL) in the presence or absence of irisin for 14 d. Neurons were stained with Hoechst 33342 (7 M) and propidium iodide (PI) (2 M) (Invitrogen). Learn more about propidium iodide, and propidium iodide containing products. We have protocols for staining using DNA binding dyes with and without antibody staining and a protocol for BrdU staining. Since propidium iodide is not permeant to live cells, it is also commonly used to detect dead cells in a population. Our labeling reagents enable researchers to create their own labeled biomolecule for use in immunochemistry, fluorescence in situ hybridization (FISH), cell tracing, receptor labeling, and cytochemistry applications as well as for probing biological structure, function, and interactions.. We also offer our fluorophores conjugated to a variety of antibodies, streptavidin, peptides, Detection of dead cells is accomplished by measuring movement of molecules either into or out of cells 5a . 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. An independent experiment from that shown in Fig. Flow Cytometric Analysis of FITC Annexin V staining. The molar ratio of tannic acid and sodium citrate (1:1) used in the AgNP synthesis was chosen to obtain AgNPs with an average size of 61 10 nm and moderate monodispersity (), given by their PDI values lower than 0.25.The surface charge of plain AgNPs was highly negative (59 6 mV), mainly due to the free COOH/OH groups of tannic acid and sodium citrate on the References and Notes. Fluorescent conjugates. Standard Cell Staining Protocol: Prior to reconstitution, spin down the vial of lyophilized reagent in a microcentrofuge to ensure the reagent is at the bottom of the vial. Thus, using Fixable Viability Dyes allows dead cells to be excluded from analysis when intracellular targets are being studied. The Invitrogen Molecular Probes Alexa Fluor 488 dyewith spectral properties and quantum yield nearly identical to those of fluorescein isothiocyanate (FITC)produces brighter, more photostable conjugates.These conjugates are ideal for imaging and other applications requiring increased sensitivity and environmentally insensitive Images were taken by microscope and dead cells were automatically counted by Axiovision 4.6 software (Carl Zeiss). View/request a protocol for this paper from Bio-protocol. When a viability dye is introduced, in this case propidium iodide, you can see that the same forward and side scatter contains both live and dead cells (Figure 3b). Jurkat cells were stained according to the protocol in the LIVE/DEAD Violet Viability/Vitality Kit. Flow Cytometric Analysis of FITC Annexin V staining. Affix the sample on glass slide (To ensure the validity of fluorescence staining, positive, negative and sample autofluorescence controls should be carried out to confirm there is no non-specific binding.) The qPCR protocol was performed for 40 cycles and each cycle consisted of denaturation at 95 C for 15 s, annealing at 60 C for 1 min, and extension at 72 C for 1 min. Standard Cell Staining Protocol: Prior to reconstitution, spin down the vial of lyophilized reagent in a microcentrofuge to ensure the reagent is at the bottom of the vial. Dead Cell Apoptosis Kit with Annexin V Alexa Fluor 488 & Propidium Iodide (PI) FC, FM: 499/521 535/617: V13241: Dead Cell Apoptosis Kit with Annexin V FITC and PI: FC: 494/518 Staining of a mixture of heat-killed and untreated Jurkat cells. Add properly diluted primary antibody to cover the sample Observe cell staining with fluorescence microscopy . 3. Propidium iodide was used as a counterstain to discriminate necrotic/ dead cells from apoptotic cells. 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. View/request a protocol for this paper from Bio-protocol. Resuspended cells were incubated with Annexin V-FITC for 15 min in the dark. Jurkat cells (Human T-cell leukemia; ATCC TIB-152) were left untreated (top panels) or treated for 4 hours with 12 M campotothecin (bottom panels). 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. The molar ratio of tannic acid and sodium citrate (1:1) used in the AgNP synthesis was chosen to obtain AgNPs with an average size of 61 10 nm and moderate monodispersity (), given by their PDI values lower than 0.25.The surface charge of plain AgNPs was highly negative (59 6 mV), mainly due to the free COOH/OH groups of tannic acid and sodium citrate on the 556417) followed by incubation with SAv-FITC (Cat. For Research Use Only. Our labeling reagents enable researchers to create their own labeled biomolecule for use in immunochemistry, fluorescence in situ hybridization (FISH), cell tracing, receptor labeling, and cytochemistry applications as well as for probing biological structure, function, and interactions.. We also offer our fluorophores conjugated to a variety of antibodies, streptavidin, peptides, Immunofluorescence staining of cells in combination with PI staining of cells for cell cycle analysis; Propidium iodide staining of cells for cell cycle analysis ; BrdU staining of cells for cell cycle analysis Fluorescent conjugates. The gasdermin E protein is shown to act as a tumour suppressor: it is cleaved by caspase 3 and granzyme B and leads to pyroptosis of cancer cells, provoking an immune response to the tumour. 3. Cells were then analyzed by flow cytometry. Cells were incubated with Annexin V-Biotin, (Cat. Propidium iodide is used as a DNA stain in flow cytometry to evaluate cell viability or DNA content in cell cycle analysis, or in microscopy to visualise Propidium iodide. Cover the cells with a glass coverslip. Typically, a membrane-impermeable dye like propidium iodide is used to identify dead or dying cells with damaged membranes and a viability dye like calcein-AM used to label live cells. Cells were then analyzed by flow cytometry. No. LDH activity Cells were incubated with FITC Annexin V in a buffer containing propidium iodide (PI) and analyzed by flow cytometry. Its selectivity for DNA allows efficient staining of nuclei with little background from the cytoplasm. Annexin V-FITC/ PI staining of AG06173 primary fibroblasts. Primary cortical neurons were treated with -syn PFF (5 g/mL) in the presence or absence of irisin for 14 d. Neurons were stained with Hoechst 33342 (7 M) and propidium iodide (PI) (2 M) (Invitrogen). 1. References and Notes. Learn more about propidium iodide, and propidium iodide containing products. References and Notes. LDH activity Unlike 7-AAD and propidium iodide, cells labeled with Fixable Viability Dyes can be washed, fixed, permabilized, and stained for intracellular antigens without any loss of staining intensity of the dead cells. Unlike 7-AAD and propidium iodide, cells labeled with Fixable Viability Dyes can be washed, fixed, permabilized, and stained for intracellular antigens without any loss of staining intensity of the dead cells. Immunofluorescence staining of cells in combination with PI staining of cells for cell cycle analysis; Propidium iodide staining of cells for cell cycle analysis ; BrdU staining of cells for cell cycle analysis Hoechst 33342 is used for specifically staining the nuclei of living or fixed cells and tissues. For reconstitution, pre-warm the kit to room temperature; add 100 l of DMSO to PI is widely used in fluorescence microscopy, confocal laser scanning microscopy, flow cytometry, and fluorometry. Images were taken by microscope and dead cells were automatically counted by Axiovision 4.6 software (Carl Zeiss). When a viability dye is introduced, in this case propidium iodide, you can see that the same forward and side scatter contains both live and dead cells (Figure 3b). Cells were incubated with FITC Annexin V in a buffer containing propidium iodide (PI) and analyzed by flow cytometry. TO-PRO-3 and TOTO-3 stains both have much greater extinction coefficients than DNA-bound propidium iodide. Cover the cells with a glass coverslip. Cells were incubated with Annexin V-Biotin, (Cat. No. 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. 10 5 cells were used for analysis. ). Left: negative control - AG6173 untreated cells. When a viability dye is introduced, in this case propidium iodide, you can see that the same forward and side scatter contains both live and dead cells (Figure 3b). 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. Fluorophore is detected via microscopy; Protocol. Propidium iodide is used as a DNA stain in flow cytometry to evaluate cell viability or DNA content in cell cycle analysis, or in microscopy to visualise Untreated cells were primarily Annexin V-Biotin and PI negative, indicating that they were viable and not undergoing apoptosis. For Research Use Only. Propidium iodide (PI) is a popular red-fluorescent nuclear and chromosome counterstain. Hoechst 33342 is used for specifically staining the nuclei of living or fixed cells and tissues. No. Jurkat cells (Human T-cell leukemia; ATCC TIB-152) were left untreated (top panels) or treated for 4 hours with 12 M campotothecin (bottom panels). 3. Dead Cell Apoptosis Kit with Annexin V Alexa Fluor 488 & Propidium Iodide (PI) Kit Contents: Contains 5 vials of annexin V, Alexa Fluor 488 conjugate (250 L per vial), 1 vial of propidium iodide (PI, 100 L), and 1 bottle of annexin binding buffer (5X solution, 50 mL). Fluorophore is detected via microscopy; Protocol. S1, A to C). a, Confocal laser scanning microscopy fluorescence imaging of glioma cells that were treated with each formulation. a, Confocal laser scanning microscopy fluorescence imaging of glioma cells that were treated with each formulation. The Invitrogen Molecular Probes Alexa Fluor 488 dyewith spectral properties and quantum yield nearly identical to those of fluorescein isothiocyanate (FITC)produces brighter, more photostable conjugates.These conjugates are ideal for imaging and other applications requiring increased sensitivity and environmentally insensitive Since propidium iodide is not permeant to live cells, it is also commonly used to detect dead cells in a population. No. Propidium iodide. 10 5 cells were used for analysis. Product Type: Apoptosis Detection Kit: Flow Cytometer Laser Lines: 488 Probes with high fluorescence quantum yield and high photostability allow detection of low-abundance biological structures with great sensitivity. Annexin V and propidium iodide (PI) staining confirmed that NK-92MI cells induced more extensive lytic death in GSDMB-expressing 293T cells than that in cells expressing other gasdermins or the empty vector (fig. Add properly diluted primary antibody to cover the sample Cover the cells with a glass coverslip. Thus, using Fixable Viability Dyes allows dead cells to be excluded from analysis when intracellular targets are being studied. Expand Your Options for Multicolor Labeling 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. 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. The original propidium iodide staining dataset was resampled to 10 m 3. 556417) followed by incubation with SAv-FITC (Cat. Primary cortical neurons were treated with -syn PFF (5 g/mL) in the presence or absence of irisin for 14 d. Neurons were stained with Hoechst 33342 (7 M) and propidium iodide (PI) (2 M) (Invitrogen). 1. Typically, a membrane-impermeable dye like propidium iodide is used to identify dead or dying cells with damaged membranes and a viability dye like calcein-AM used to label live cells. 2.2 If propidium iodide was added, analyze PI staining by the phycoerythrin emission signal detector (usually FL2). Probes with high fluorescence quantum yield and high photostability allow detection of low-abundance biological structures with great sensitivity. Scale bars, 100 m. HUVECs with compromised DNA integrity and/or membranes were indicated by propidium iodide (PI) staining with a 22 h treatment (Fig. We have protocols for staining using DNA binding dyes with and without antibody staining and a protocol for BrdU staining. 2.2 If propidium iodide was added, analyze PI staining by the phycoerythrin emission signal detector (usually FL2). 1d and Supplementary Fig. LDH activity PI is widely used in fluorescence microscopy, confocal laser scanning microscopy, flow cytometry, and fluorometry. 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