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Csu 22

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The CSU-22 is a laboratory equipment manufactured by Nikon. It is a compact and versatile upright fluorescence microscope designed for a wide range of applications in life science research. The CSU-22 provides high-speed confocal imaging capabilities, allowing for real-time observation of live cell dynamics and processes.

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Lab products found in correlation

Prolong gold, by Thermo Fisher Scientific (1 mentions) Imagem emccd camera, by Hamamatsu Photonics (1 mentions) Lsm 5 pascal confocal microscope, by Zeiss (1 mentions) Cryostat, by Leica (1 mentions) Ti e inverted microscope, by Nikon (1 mentions) Fluoromount g, by Clinisciences (1 mentions) Anti kiss1, by Abcam (1 mentions) Ti inverted fluorescence microscope, by Nikon (1 mentions) Anti gfp antibody, by Novus Biologicals (1 mentions) Anti erα antibody, by Merck Group (1 mentions) Te2000 microscope, by Nikon (1 mentions)

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Nikon Ti inverted CSU-22 spinning disc confocal microscope (2 citations)

6 protocols using csu 22

1

Live-Cell Spinning Disc Confocal Imaging

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Live-cell spinning disc confocal imaging was carried out using Yokagawa CSU-22 confocal Nikon Ti microscope with a 60x or 100× 1.4 NA Plan Apo VC oil objective and a temperature-, humidity- and CO2-controlled Okolab enclosure Bold Line incubator. 488-nm, 568-nm and 640-nm Coherent OBIS lasers were used as light sources. Time-lapse images were acquired with an Evolve Delta EMCCD camera (Photometrics) driven by Micro-Manager 1.4 or Nikon Elements 4.5. Cells were imaged every 2–30 s for up to 30 min in DMEM without phenol red supplemented with 30 mM HEPES, pH 7.4 (UCSF Cell Culture Facility). Agonists or antagonists were added by bath application during acquisition as indicated in the figure legends. For immunofluorescence imaging, cells were cells grown on coverslips and fixed using 4% formaldehyde in PBS. Cells were permeabilized with 0.05% saponin and 1% BSA in PBS and incubated with primary (1:1000) and secondary (1:1000) antibodies. Specimens were mounted using ProLong Gold (Thermo Fisher).
Stoeber M., Jullié D., Lobingier B.T., Laeremans T., Steyaert J., Schiller P.W., Manglik A, & von Zastrow M. (2018). A genetically encoded biosensor reveals location bias of opioid drug action. Neuron, 98(5), 963-976.e5.
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2

Live Cell Imaging of Artificial LDs

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Live cell imaging was performed as described using high numerical aperture 60× or 100× objectives (Wang et al., 2016 ). For in vitro experiments, imaging was performed with a spinning disk confocal (Yokogawa CSU22) set up on a Nikon Eclipse Ti inverted microscope. Illumination was performed with 488 and 561 nm laser lines, and detection with an imagEM EM-CCD camera (Hamamatsu). TG-loaded GUVs: imaging was performed with a 60X ApoTIRF 1.49 NA objective (Nikon). Artificial LDs: LDs float at the top of the observation chamber and were imaged using a lower magnification/ longer working distance objective (Plan Apo VC 20X, 0.75 NA, Nikon).
Prévost C., Sharp M.E., Kory N., Lin Q., Voth G.A., Farese RV J.r, & Walther T.C. (2018). Mechanism and Determinants of Amphipathic Helix-Containing Protein Targeting to Lipid Droplets. Developmental cell, 44(1), 73-86.e4.
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3

Immunolabeling of Cryosectioned Samples

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14-micron sections were cut on a cryostat (Leica) and stored at −80 °C until use. Sections were permeabilized with Triton 0.3% for 15 min. After a 15-min quenching step with 50 mM NH4Cl, sections were blocked either with 5% horse or donkey serum, 0.1% Triton X-100 in PBS, or with 5% fetal bovine serum, 1% BSA in PBS when the staining involved anti-Prominin-1 antibody. Sections were incubated overnight at 4 °C with primary antibodies in blocking solution, then with fluorescence-conjugated secondary antibodies, followed by DAPI or Sytox Green staining. Slices were mounted in Fluoromount-G (Clinisciences). Images were acquired using a LSM5 Pascal confocal microscope (Zeiss), a spectral C1si confocal microscope (Nikon), or a spinning disk confocal (Yokogawa CSU22) on a Nikon Ti-E inverted microscope.
Rakotomamonjy J., Brunner M., Jüschke C., Zang K., Huang E.J., Reichardt L.F, & Chenn A. (2017). Afadin controls cell polarization and mitotic spindle orientation in developing cortical radial glia. Neural Development, 12, 7.
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4

Quantifying Cell-Associated Bacterial Aggregation

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For experiments in which we quantified the efficiency of cell-associated aggregation, images were acquired on a Nikon Ti-E microscope equipped with a Yokagawa CSU22 spinning disk confocal with a Nikon Plan Fluor 40×/1.30 oil objective. Using Micro-Manager software to pre-program a region of ∼5*106 square microns (342 fields) in the center of each specimen, we quantified the number of cell-associated aggregates (defined as groupings of ≥10 bacteria) and normalized them to results from same-day infection with PAK control. Results are reported for three or more independent experiments.
Tran C.S., Rangel S.M., Almblad H., Kierbel A., Givskov M., Tolker-Nielsen T., Hauser A.R, & Engel J.N. (2014). The Pseudomonas aeruginosa Type III Translocon Is Required for Biofilm Formation at the Epithelial Barrier. PLoS Pathogens, 10(11), e1004479.
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5

Immunohistochemical Analysis of ER, GFP, and Kisspeptin

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Immunohistochemistry was performed on cryosections (20 µm) collected from brains fixed in 4% paraformaldehyde using standard procedures. ERα staining used rabbit polyclonal anti-ERα antibody (EMD Millipore, Billerica, MA cat # C1355) or mouse monoclonal ERα antibody (Abcam, Cambridge cat # 93021) at a dilution of 1:1000 or 1:100, respectively. GFP staining used anti-GFP antibody (Novus Biologicals, Littleton, CO cat # NB100-1614) at 1:2500. Kisspeptin staining used rabbit polyclonal anti-KISS1 at a dilution of 1:200 (Abcam, Cambridge cat # ab19028). Confocal images were taken using a Nikon Ti inverted fluorescence microscope with CSU-22 spinning disk confocal.
Herber C.B., Krause W.C., Wang L., Bayrer J.R., Li A., Schmitz M., Fields A., Ford B., Zhang Z., Reid M.S., Nomura D.K., Nissenson R.A., Correa S.M, & Ingraham H.A. (2019). Estrogen signaling in arcuate Kiss1 neurons suppresses a sex-dependent female circuit promoting dense strong bones. Nature Communications, 10, 163.
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6

Imaging and Quantification of Ire1 Variants

Cited 1 time
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All imaging and quantitation of images were performed, as described (Aragón et al., 2009 (link)). In brief, samples for microscopy were taken from yeast that was kept in early log-phase for at least 16 hr in 2× synthetic media before imaging. Microscopy of laser-excited mCherry or GFP was performed with a Yokogawa CSU-22 spinning disc confocal on a Nikon TE2000 microscope, controlled with µmanager and ImageJ. Images were captured with a 100×/1.4 NA Plan Apo objective on a Cascade II EMCCD and selected for analysis to contain significant signal above background but no saturated pixels. For display, images were processed in ImageJ and Adobe Photoshop such that the linear range of signals was comparable. Foci of Ire1 variants were determined and the co-localization index for U1A–GFP decorated SpRU1A, SL-PGK-3′ hac1U1A, or SpR Δ3′ BEU1A mRNA recruited to those foci was scored by using a customized MatLab script, as described (Aragón et al., 2009 (link)).
van Anken E., Pincus D., Coyle S., Aragón T., Osman C., Lari F., Gómez Puerta S., Korennykh A.V, & Walter P. (2014). Specificity in endoplasmic reticulum-stress signaling in yeast entails a step-wise engagement of HAC1 mRNA to clusters of the stress sensor Ire1. eLife, 3, e05031.
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