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Sp5 dm microscope

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The SP5 DM microscope is a confocal laser scanning microscope designed for high-resolution imaging and analysis of biological samples. It features a modular design, allowing for customization to meet specific research needs. The SP5 DM provides advanced imaging capabilities, including multi-channel fluorescence detection, z-stacking, and time-lapse imaging.

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Lab products found in correlation

Prolong diamond antifade mountant, by Thermo Fisher Scientific (2 mentions) Las x software, by Leica (2 mentions) 4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (2 mentions) Anti mtco2, by Abcam (2 mentions) Anti tom20, by Cell Signaling Technology (1 mentions) Anti lamp1, by Cell Signaling Technology (1 mentions)

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SP5 microscope (176 citations) SP5 DM confocal microscope (6 citations) SP5 DM (3 citations)

9 protocols using sp5 dm microscope

1

Quantitative Analysis of Synaptic Proteins

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Immunolabeled hippocampal neurons were visualized by using a Leica SP5 DM microscope with a 63x magnification objective and analyzed through the ImageJ software (imagej.nih.gov). Confocal z-stack images were acquired in 17 (wt) and 14 (m) random locations within the coverslips of three independent transfections. For SYN1 quantification, individual cells (42 wt and 44 m cells) were circled based on their fluorescent signals and the area and integrated mean intensity was then calculated in ImageJ. The corrected total cell fluorescence (CTCF) was calculated as Integrated Density (area of selected cells x mean fluorescence of background readings)55 (link). The length of the neurites was measured by using the ImageJ software as described elsewhere13 (link).
Darvish H., Azcona L.J., Tafakhori A., Mesias R., Ahmadifard A., Sanchez E., Habibi A., Alehabib E., Johari A.H., Emamalizadeh B., Jamali F., Chapi M., Jamshidi J., Kajiwara Y, & Paisán-Ruiz C. (2020). Phenotypic and genotypic characterization of families with complex intellectual disability identified pathogenic genetic variations in known and novel disease genes. Scientific Reports, 10, 968.
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2

Immunostaining of Macrophages After Bacterial Stimulation

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Macrophages were plated on glass coverslips. After bacterial stimulation, coverslips were washed with PBS, fixed in 2% PFA, and quenched in 50mM NH4Cl. Cells were then permeabilized with 0.2% Triton X-100 in PBS before blocking with 10% FBS PBS and incubation with appropriate antibodies prepared in 10% FBS PBS. When indicated, Phalloidin Alexa 647 was used together with secondary fluorescent antibodies to stain Actin and delineate the cell area. Lastly, cells were stained with DAPI. Coverslips mounted with ProLong Diamond Antifade Mountant (Invitrogen). Images acquired on a Leica SP5 DM microscope with a 63x/1.4 NA oil immersion objective. The partial nuclear staining pattern noted in Extended Data Figure 4 has been reported60 (link) and is ASC-specific given its absence in Pycard–/– macrophages.
Moretti J., Jia B., Hutchins Z., Roy S., Yip H., Wu J., Shan M., Jaffrey S.R., Coers J, & Blander J.M. (2022). Caspase-11 Interaction with NLRP3 Potentiates the Noncanonical Activation of the NLRP3 Inflammasome. Nature immunology, 23(5), 705-717.
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3

Immunostaining of Macrophages After Bacterial Stimulation

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Macrophages were plated on glass coverslips. After bacterial stimulation, coverslips were washed with PBS, fixed in 2% PFA, and quenched in 50mM NH4Cl. Cells were then permeabilized with 0.2% Triton X-100 in PBS before blocking with 10% FBS PBS and incubation with appropriate antibodies prepared in 10% FBS PBS. When indicated, Phalloidin Alexa 647 was used together with secondary fluorescent antibodies to stain Actin and delineate the cell area. Lastly, cells were stained with DAPI. Coverslips mounted with ProLong Diamond Antifade Mountant (Invitrogen). Images acquired on a Leica SP5 DM microscope with a 63x/1.4 NA oil immersion objective. The partial nuclear staining pattern noted in Extended Data Figure 4 has been reported60 (link) and is ASC-specific given its absence in Pycard–/– macrophages.
Moretti J., Jia B., Hutchins Z., Roy S., Yip H., Wu J., Shan M., Jaffrey S.R., Coers J, & Blander J.M. (2022). Caspase-11 Interaction with NLRP3 Potentiates the Noncanonical Activation of the NLRP3 Inflammasome. Nature immunology, 23(5), 705-717.
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4

Confocal Microscopy Immunolabeling Analysis

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Immunolabeling was visualized using a Leica SP5 DM microscope with a 63 × magnification objective. Confocal z-stack images were acquired in six random locations within coverslip per condition while blinded to genotype. Images were then analyzed using ImageJ (http://imagej.nih.gov/ij/) [Schneider, et al., 2012 (link)].
Sanchez E., Darvish H., Mesias R., Taghavi S., Firouzabadi S.G., Walker R.H., Tafakhori A, & Paisán-Ruiz C. (2016). Identification of a Large DNAJB2 Deletion in a Family with Spinal Muscular Atrophy and Parkinsonism. Human mutation, 37(11), 1180-1189.
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5

Dendritic Cell Stimulation Assay

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DC were seeded onto Alcian blue-treated glass coverslips and stimulated with either streptavidin beads or streptavidin beads conjugated to biotinylated LPS for 3 hr. Coverslips were fixed and permeabilized prior to staining. Images were acquired on a Leica SP5 DM microscope with a 633/1.4 NA oil immersion objective.
Nair-Gupta P., Baccarini A., Tung N., Seyffer F., Florey O., Huang Y., Huang M., Overholtzer M., Roche P.A., Tampé R., Brown B.D., Amsen D., Whiteheart S.W, & Blander J.M. (2014). TLR Signals Induce Phagosomal MHC-I Delivery from the Endosomal Recycling Compartment to Allow Cross-Presentation. Cell, 158(3), 506-521.
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6

Phagocytosis Imaging in Dendritic Cells

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DCs were seeded onto Alcian blue-treated glass coverslips and stimulated or not with phagocytic cargo. Streptavidin beads with or without conjugation to biotinylated LPS were added to DCs (1DC:2beads) for 3h. Coverslips were fixed and permeabilized prior to staining. Images were acquired on a Leica SP5 DM microscope with a 633/1.4 NA oil immersion objective.
Barbet G., Nair-Gupta P., Schotsaert M., Yeung S.T., Moretti J., Seyffer F., Metreveli G., Gardner T., Choi A., Tortorella D., Tampé R., Khanna K.M., García-Sastre A, & Blander J.M. (2021). TAP dysfunction in dendritic cells enables non-canonical cross-presentation for T cell priming. Nature immunology, 22(4), 497-509.
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7

Phagocytosis Imaging in Dendritic Cells

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DCs were seeded onto Alcian blue-treated glass coverslips and stimulated or not with phagocytic cargo. Streptavidin beads with or without conjugation to biotinylated LPS were added to DCs (1DC:2beads) for 3h. Coverslips were fixed and permeabilized prior to staining. Images were acquired on a Leica SP5 DM microscope with a 633/1.4 NA oil immersion objective.
Barbet G., Nair-Gupta P., Schotsaert M., Yeung S.T., Moretti J., Seyffer F., Metreveli G., Gardner T., Choi A., Tortorella D., Tampé R., Khanna K.M., García-Sastre A, & Blander J.M. (2021). TAP dysfunction in dendritic cells enables non-canonical cross-presentation for T cell priming. Nature immunology, 22(4), 497-509.
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8

Mitochondrial, Lysosomal, and Nuclear Staining

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MDMs were seeded at a density of 20,000 cells/well. Then mitochondria were stained using anti-MTCO2 (Abcam) or anti-TOM20 (Cell Signaling Technology). Lysosomes were stained with anti-LAMP1 (Cell Signaling Technology), and nuclei were stained with DAPI (Acros Organics). Fluorescence microscopy was conducted with FV10-ASW4.1 viewer and Leica SP5 DM microscope and analyzed with LASX software (Leica).
Khan S., Raj D., Sahu S., Naseer A., Singh N.C., Kumari S., Ishteyaque S., Sharma J., Lakra P., Mugale M.N., Trivedi A.K., Srivastava M., Chandra T., Bhosale V., Barthwal M.K., Gupta S.K., Mitra K., Nazir A., Ghoshal U.C, & Lahiri A. (2023). CLUH functions as a negative regulator of inflammation in human macrophages and determines ulcerative colitis pathogenesis. JCI Insight, 8(11), e161096.
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9

Visualizing Mitochondria in Human Colon Tissue

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Human colon tissue (fixed in tissue-freezing medium) was cut in 4 μm thick sections, and mitochondria were stained using anti-MTCO2 (Abcam); nuclei were stained with DAPI (Acros Organics). Confocal fluorescence microscopy was conducted with Leica SP5 DM microscope with LASX software (Leica).
Khan S., Raj D., Sahu S., Naseer A., Singh N.C., Kumari S., Ishteyaque S., Sharma J., Lakra P., Mugale M.N., Trivedi A.K., Srivastava M., Chandra T., Bhosale V., Barthwal M.K., Gupta S.K., Mitra K., Nazir A., Ghoshal U.C, & Lahiri A. (2023). CLUH functions as a negative regulator of inflammation in human macrophages and determines ulcerative colitis pathogenesis. JCI Insight, 8(11), e161096.
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