Following plating on gelatin-coated Lab-Tek I chambers, cells were maintained in phenol red-deprived DMEM-F12 (supplemented with low serum growth supplement and 10% FBS) for epifluorescence video microscopy experiments. For bright field video microscopy, cells were observed over a 30 min period using a Zeiss Axiovert 200M equipped with a Coolsnap camera (Roper Scientific, Trenton, NJ, USA). Cells were maintained at 37°C in 5% CO2 atmosphere using a temperature-controlled chamber (PECON, Erbach, Germany). Data were acquired using MetaMorph software (Roper Scientific). Timespan between image capture was 4 s. Confocal microscopy images of fixed cells were collected with a Zeiss (Oberkochen, Germany) LSM710 Meta confocal microscope using either a ×63 Plan Apochromat objective (oil immersion, 1.40 NA, DIC) at a 132 nm.pixel-1 resolution, leading to a slight XY oversampling. Optical slices were spaced by 1 μm so as to limit Z oversampling. Fluorescence filters were chosen and tested for each combination of plasmid construction and fluorescent dye so as to abolish spectral overlap. Zen software program was used to acquire images.
Coolsnap camera
Manufactured by Teledyne
Sourced in United States
The CoolSNAP camera is a high-performance digital imaging device designed for scientific and industrial applications. It features a cooled CCD sensor that provides low noise and high sensitivity, enabling accurate and reliable image capture. The camera is capable of capturing images with high resolution and bit depth, making it suitable for a variety of imaging tasks.
Automatically generated - may contain errors
Lab products found in correlation
Eclipse 800 microscope, by Nikon (3 mentions)
Metamorph software, by Universal Imaging (2 mentions)
Plan apochromat 100, by Zeiss (2 mentions)
Lsm 510 meta, by Zeiss (2 mentions)
Metamorph software, by Teledyne (1 mentions)
Axiovert 200m, by Teledyne (1 mentions)
Lsm 710 meta, by Zeiss (1 mentions)
Inverted microscope, by Nikon (1 mentions)
Fluorodish culture dishes, by World Precision Instruments (1 mentions)
Metamorph imaging system, by Universal Imaging (1 mentions)
Metamorph, by Adobe (1 mentions)
Fm4 64fx, by Thermo Fisher Scientific (1 mentions)
4 6 diamino 2 phenylindole dapi, by Thermo Fisher Scientific (1 mentions)
D300 camera, by Nikon (1 mentions)
Eclipse ti system, by Nikon (1 mentions)
Alexa flour 488, by Thermo Fisher Scientific (1 mentions)
Rabbit anti flag, by Merck Group (1 mentions)
Alexa fluor 594, by Thermo Fisher Scientific (1 mentions)
Mouse anti flag m2, by Merck Group (1 mentions)
Axioplan 2 fluorescence microscope, by Zeiss (1 mentions)
11 protocols using coolsnap camera
1
Live-cell and confocal microscopy of cells
2
Cellular Morphology Assessment via Microscopy
3
Live Sperm Imaging and Analysis
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Live spermatozoa were imaged using a Nikon Eclipse 800 microscope (Nikon Instruments Inc.) with Cool Snap camera (Roper Scientific, Tuscon, AZ, USA) and MetaMorph software (Universal Imaging Corp., Downington, PA, USA). Images were adjusted for contrast and brightness in Adobe Photoshop CS5 (Adobe Systems, Mountain View, CA) to match the fluorescence intensities viewed through the microscope eyepieces.
4
Quantifying Autophagy and Mitochondrial Potential
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Live control- or rottlerin treated GFP-LC3 mPaSC grown on fluorodish culture dishes (World Precision Instrument) were imaged with an inverted Nikon Eclipse TE2000-S fluorescence microscope equipped with a CoolSnap camera (Roper Scientific). For quantification of GFP-LC3 puncta, cells displaying >10 brightly fluorescent GFP-LC3 puncta were counted as positive. For assessment of mitochondria membrane potential, treated GFP-LC3 cells were loaded with MitoTracker Red CMXRos and imaged as indicated above. Images were analyzed using the MetaMorph imaging system (Universal Imaging Corporation).
5
Imaging and Quantifying Yeast Cellular Features
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Log-phase yeast were imaged in minimal selective medium at room temperature with a Plan-Apochromat 100×/1.40 numerical aperture oil immersion objective lens on an Axioplan 2 fluorescence microscope (Carl Zeiss, Jena, Germany). Images were taken with a CoolSNAP camera (Roper Scientific; Tucson, AZ) using MetaMorph 7.7 software (MDS Analytical Technologies, Sunnyvale, CA) and adjusted using MetaMorph and Photoshop CS5 (Adobe, San Jose, CA). Exposure times varied from 100 ms to 3 s based on the protein tagged with GFP or RFP but were kept the same within a given experiment. When the FM4-64 (T-3166; Life Technologies) lipophilic dye was used, cells were incubated with the dye for 1 h in minimal medium, washed once, and then grown another hour in minimal medium before imaging. Cellular features were quantified by manually scoring images.
6
Visualizing Cell Membrane and Nucleoid in Bacterial Strains
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Strains, grown under appropriate conditions as described in the text, were stained with FM4-64FX (Molecular Probes) to visualize the cell membrane and with 4’,6-diamino-2-phenylindole (DAPI, Molecular Probes) to visualize the nucleoid. Cells were deposited on slides covered with 1.2% agarose in minimal medium [75 (link)]. Cells were examined using a Leica DMRA2 microscope and a COOLSNAP camera (Roper Scientific USA). Images were captured and analyzed using Metamorph V6.3r5.
7
Microscopic Imaging of Plant Seedlings
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Bright-field and fluorescence images of seedlings were collected with a Leica MZFLIII microscope and CoolSNAP camera (Roper Scientific) or Nikon D300 camera using Qcapture software. Confocal images were acquired on Leica SP5 or Zeiss LSM510META, and long-term imaging was performed under perfluorodecalin (F2 Chemicals) in Carolina Gel (Blades Biological) chambers. Immunoelectron microscopy was performed as previously described (Dettmer et al., 2006 (link)) on ultrathin thawed cryosections of formaldehyde-fixed lateral roots.
8
Apoptosis Induction by BSE in A375 Cells
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To investigate whether BSE induced apoptosis in A375 cells, we analyzed the dose dependent effect of BSE (1.5–1.9 mg/mL) on cellular morphology after 24 h treatment using the Annexin-V-FLUOS Staining kit for detection of apoptotic and necrotic cells according to vendor's protocol (Roche Diagnostics Corporation, Indianapolis, IN). This kit uses a dual-staining protocol in which the apoptotic cells are stained with Annexin-V (green fluorescence), and the necrotic cells are stained with PI (red fluorescence). Fluorescence was visualized using a Nikon Eclipse Ti system (Nikon Instruments Inc.; Tokyo, Japan), and digital images were captured with an attached CoolSNAP camera (Roper Scientific, Trenton, NJ) linked to a computer.
9
Immunofluorescent Labeling of Transfected Cells
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Twenty hours post-transfection, cells were washed twice with PBS, and fixed in 4% paraformaldehyde (PFA) solution (4% PFA, 4% sucrose in PBS) for 20 min. Cells were permeabilized for 1 min in PBS containing 0.1% TX-100, washed thrice in PBS, and blocked with PBS +3% BSA for 60 min before incubating with primary antibodies (mouse anti-FLAG-M2, 1:500; rabbit anti-FLAG (Sigma), 1:200; or mouse anti-GM130 (BD Biosciences; San Jose, CA), 1:200) for 1 hr. Coverslips were washed thrice and incubated with secondary antibodies (goat anti-mouse Alexa Flour 488 and goat anti-rabbit Alexa Fluor 594 (Life Technologies), 1:1000 each) for an hour. Coverslips were washed with PBS and mounted on glass slides with ProLong Gold Antifade Mountant containing DAPI. Cells were observed with an Axioplan 2 fluorescence microscope (Carl Zeiss; Oberkochen, Germany) using a Plan-Apochromat 100× 1.40 NA oil immersion objective lens. Images were acquired with a CoolSNAP camera (Roper Scientific; Planegg, Germany) using YFP, GFP, and Texas Red filters and MetaMorph 7.7 software (MDS analytical Technologies; Toronto, ON), and adjusted using Metamorph 7.7.
10
Sperm Mitochondrial Sheath Measurement
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Fixed sperm were sedimented on pol-L-lysine coverslips, permeabilized for 40 min in PBS + 0.1% TrX, then incubated for 10 min with 1:1000 AGG probe. Following probe incubation, coverslips were washed in PBS-TrX and images acquired in 10 random fields per boar using a 100 × oil lens on a Nikon Eclipse 800 microscope (Nikon Instruments Inc., Melville, NY, USA) with Cool Snap camera (Roper Scientific, Tuscon, AZ, USA) and MetaMorph software (Universal Imaging Crop., Downington, PA, USA). AGG fluorescence was acquired in the green channel at 500 millisecond acquisition time. The mitochondrial sheath lengths of 50 intact, grossly morphologically normal spermatozoa, were measured and recorded using the tape measure tool in MetaMorph (Version 7, Universal Imaging Corp., Downington, PA, USA).
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