imaris software colocalization package  (Oxford Instruments)

Journal: American Journal of Physiology - Regulatory, Integrative and Comparative Physiology

Article Title: Maternal food restriction modulates cerebrovascular structure and contractility in adult rat offspring: effects of metyrapone

doi: 10.1152/ajpregu.00436.2013

Figure Lengend Snippet: Colocalization of smooth muscle α-actin with myosin heavy-chain isoforms. Concentric rings of rat middle cerebral arteries were immunostained to quantify colocalization of smooth muscle α-actin with one of three myosin heavy-chain isoforms. Shown here are representative sections from all three experimental groups immunostained for colocalization of smooth muscle α-actin with the nonmuscle isoform of myosin heavy chain.

Article Snippet: The degree of colocalization between the two markers was determined using the colocalization coefficient calculated within the FluoView software package (Olympus) because this measure was independent of absolute marker intensities.

Techniques:

Journal: American Journal of Physiology - Regulatory, Integrative and Comparative Physiology

Article Title: Maternal food restriction modulates cerebrovascular structure and contractility in adult rat offspring: effects of metyrapone

doi: 10.1152/ajpregu.00436.2013

Figure Lengend Snippet: Effects of maternal food restriction and metyrapone on colocalization of smooth muscle α-actin with myosin heavy-chain isoforms. Colocalization with smooth muscle α-actin was with nonmuscle myosin heavy chain (NM) was significantly greater in the MFR group than in either the Control group or the MFR + Metyrapone group (*P < 0.05). Colocalization of smooth muscle α-actin with the SM1 myosin isoform was also significantly greater in the MFR group than in the Control group (*P < 0.05), but the SM1 colocalization value in the MFR + Metyrapone did not differ significantly from either the Control or the MFR groups. For colocalization of smooth muscle α-actin with the SM2 myosin isoform, the values in the Control and MFR groups were similar, but the value in the MFR + Metyrapone group was significantly greater than in either the Control or MFR groups (*P < 0.05). Error bars indicate means ± SE for n = 5 for each mean.

Article Snippet: The degree of colocalization between the two markers was determined using the colocalization coefficient calculated within the FluoView software package (Olympus) because this measure was independent of absolute marker intensities.

Techniques:

Journal: Cell Host & Microbe

Article Title: HIV-1 Assembly: Viral Glycoproteins Segregate Quantally to Lipid Rafts that Associate Individually with HIV-1 Capsids and Virions

doi: 10.1016/j.chom.2008.04.004

Figure Lengend Snippet: Confocal Microscopy of Transfected MAGI-CCR5 Cells Revealed that GP Did Not Colocalize with Env, and This Pattern Did Not Change in the Presence or Absence of Gag (A) MAGI-CCR5 cells were transfected with myc- or HA-Env expression vectors and tagged within the V1 region (see ), which showed colocalization and served as a positive control (left panels). MAGI-CCR5 cells were transiently transfected with GP and HA-Env-expression vectors (right panels). Immunofluorescent staining was performed 40 hr later. Z stack sectioning of the cells was collected and analyzed by Imaris software to obtain 3D reconstruction and colocalization statistics. (B) MAGI-CCR5 cells were transiently transfected with Env, GP, or double expression plasmid (Env/GP) in the presence or absence of packaging vector pCMVΔR8.2, which contains Gag. Immunofluorescent staining was performed 40 hr later. Z stack sectioning of the cells was collected and analyzed by Imaris software.

Article Snippet: The fraction of one channel volume colocalized with another channel volume, and the Pearson channel correlation in colocalized volume was calculated for cytoplasm, plasma membrane, and whole cell using Imaris software colocalization package.

Techniques: Confocal Microscopy, Transfection, Expressing, Positive Control, Staining, Software, Plasmid Preparation

Journal: Cell Host & Microbe

Article Title: HIV-1 Assembly: Viral Glycoproteins Segregate Quantally to Lipid Rafts that Associate Individually with HIV-1 Capsids and Virions

doi: 10.1016/j.chom.2008.04.004

Figure Lengend Snippet: Lack of Colocalization of Heterologous Envelope Proteins, Env and GP, in Producer Cells Confocal microscopy revealed that GP did not colocalize with Env. MAGI-CCR5 cells were transiently transfected with packaging vector pCMVΔR8.2 and GP- and HIV-Env ADA -expressing vectors. Immunofluorescence staining was performed 40 hr later. Z stack sectioning of the cells was collected and analyzed by Imaris software to obtain iso-surface models. (A) Expanded view of a thick section of a single cell is presented here as an iso-surface model depicting Env, GP, and Gag distribution in the vicinity of the plasma membrane. (B)–(D) depict two of the three viral proteins presented in (A) for clear visual comparisons. Descriptions of the method used to identify membrane versus cytoplasmic proteins with Imaris software are described further in the and Figure S4 .

Article Snippet: The fraction of one channel volume colocalized with another channel volume, and the Pearson channel correlation in colocalized volume was calculated for cytoplasm, plasma membrane, and whole cell using Imaris software colocalization package.

Techniques: Confocal Microscopy, Transfection, Plasmid Preparation, Expressing, Immunofluorescence, Staining, Software, Membrane

Journal: Cell Host & Microbe

Article Title: HIV-1 Assembly: Viral Glycoproteins Segregate Quantally to Lipid Rafts that Associate Individually with HIV-1 Capsids and Virions

doi: 10.1016/j.chom.2008.04.004

Figure Lengend Snippet: Distribution of Env and Gag in HIV-Infected Cells (A) Confocal imaging confirms the expression of Env in discrete locations intracellularly, largely separate from Gag in MAGI-CCR5 and αCD3/CD28 stimulated human CD4 lymphocytes. MAGI-CCR5 or human CD4 cells were infected with HIV-1 89.6 and analyzed by immunofluorescent staining 48 hr later. Approximately one percent of cells were infected by this criteria. (B) Z stack sectioning of the cells was collected and analyzed by Imaris software to obtain iso-surface models as in Figure 5 . A thick section of a single cell is presented here as iso-surface model for each cell type depicting Env and Gag distribution within the cytoplasm and plasma membrane as well as areas of colocalization.

Article Snippet: The fraction of one channel volume colocalized with another channel volume, and the Pearson channel correlation in colocalized volume was calculated for cytoplasm, plasma membrane, and whole cell using Imaris software colocalization package.

Techniques: Infection, Imaging, Expressing, Staining, Software, Membrane

Journal: The Journal of Cell Biology

Article Title: A Novel 14-Kilodalton Protein Interacts with the Mitogen-Activated Protein Kinase Scaffold Mp1 on a Late Endosomal/Lysosomal Compartment

doi:

Figure Lengend Snippet: Interaction between p14 and MP1 in vivo II: colocalization and comislocalization. HeLa cells were transiently cotransfected with X-p14 (top and middle panel) or X-p14–CAAX (bottom panel) and MP1-myc 6 . After 24 h incubation, the cells were fixed and processed for indirect immunofluorescence using anti–LAMP-1, anti-Xpress and anti-myc antibodies. Bars, 10 μm.

Article Snippet: Confocal images were obtained with a Leica TCS NT confocal microscope and processed using the Imaris and colocalization software packages (Bitplane AG) after deconvolution using measured point-spread functions with the Huygens software (Scientific Volume Imaging).

Techniques: In Vivo, Incubation, Immunofluorescence