Live-cell imaging was essentially performed as described (Barrales et al., 2016 (link)). In brief, cells were grown overnight in rich medium (YES) to the logarithmic phase (OD600 = 0.4–0.6). Before imaging, cells were attached to lectin (Sigma) coated glass-bottom dishes containing a microwell (MatTek). Cells were imaged using a Zeiss AxioObserver Z1 confocal spinning disk microscope with an EMM-CCD camera (Photometrics, Evolve 512) through a Zeiss Alpha Plan/Apo ×100/1.46 oil DIC M27 objective lens. Z-stacks were obtained at focus intervals of 0.4 μm. FiJi/ImageJ software was used to count the number of foci in the yeast cells.
Evolve 512
Manufactured by Zeiss
Sourced in Germany
The Evolve 512 is a scientific imaging camera designed for a wide range of microscopy applications. It features a 512 x 512 pixel sensor and provides high-resolution, low-noise imaging capabilities.
Automatically generated - may contain errors
Lab products found in correlation
Micro well, by MatTek (3 mentions)
Alpha plan apo 100 1.46 oil dic m27 objective, by Zeiss (2 mentions)
Axio observer d1 microscope, by Zeiss (2 mentions)
Janeliafluor 646 halotag ligand, by Promega (2 mentions)
Alpha plan apochromat 100 1.46 oil immersion objective, by Zeiss (2 mentions)
Janelia fluor 549 halotag ligand, by Promega (2 mentions)
Axio observer z1, by Zeiss (2 mentions)
Axio observer z, by Zeiss (1 mentions)
Axiocam mrm, by Zeiss (1 mentions)
Agarose, by Thermo Fisher Scientific (1 mentions)
Zen blue, by Zeiss (1 mentions)
Sp8x wll microscope, by Leica (1 mentions)
6 protocols using evolve 512
1
Live-Cell Imaging of Yeast Foci
2
Live-Cell Single-Molecule Tracking with Janelia Fluor Dyes
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
An Axio Observer D1 Microscope equipped with an Alpha Plan-Apochromat 100 × /1.46 Oil immersion Objective (Zeiss, Germany) and an Evolve 512 × 512 EMCCD camera with pixel size 16.0 μm was used in live-cell SMT experiments. For tracking experiments, an additional 2.5 × magnification was equipped on the emission pathway. A Solid-state LaserStack (3i) was used to excite Janelia Fluor® 549 HaloTag® Ligand (Promega; GA1110) at 552 nm, and Janelia Fluor® 646 HaloTag® Ligand (Promega; GA1120) at 640 nm, respectively. To avoid stray-light reflection and reduce background from cell auto-fluorescence, the HILO illumination model was used. A Brightline® single-band laser filter set (Semrock; excitation filter: FF01–561/14, emission filter: FF01–609/54, and dichroic mirror: Di02-R561–25 3 36) was used for the excitation and emission spectra of JF549 and HaloTag® TMR ligand. A Brightline® single-band laser filter set (Semrock; excitation filter: BLP01–635R-25, emission filter: FF01–640/14–25, and dichroic mirror: Di02-R635–25 3 36) for the excitation and emission spectra of JF646. To filter the excitation wavelength, a TIRF laser microscope cube (3i) was used. The microscope and EMCCD camera were controlled by the computer via SlideBook 6.0 software.
3
Imaging and Tracking Microglial Dynamics
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Images were taken with a Cell Observer CSU-X1 Yokogawa Spinning Disk, AxioCam MRm and Evolve 512 (Zeiss). For in vivo imaging 1.5dpf embryos or 3dpf/5dpf larvae were anesthetized in E3 1xTricaine (80 mg/l) (Pharmaq Ltd) and were mounted in 1.5% agarose (Invitrogen) dissolved in E3 1xTricaine. Embedded embryos were covered with E3 1xTricaine. Brightness and contrast were adjusted using Zen blue (Zeiss) and ImageJ. Image stitching was performed with the ImageJ plugin Image stitching [69 (link)] and microglia were tracked with the ImageJ plugin mTrackJ [70 (link)].
4
Live-Cell Single-Molecule Tracking with Janelia Fluor Dyes
5
Live-cell Imaging of Cellular Foci
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Live-cell imaging was essentially performed as described38 (link). In brief, cells were grown overnight on rich medium (YES) to the logarithmic phase. Prior to imaging, cells were attached with lectin (Sigma) to glass-bottom dishes with a micro well (MatTek). Cells were imaged on a Zeiss AxioObserver Z1 confocal spinning disk microscope with an EMM-CCD camera (Photometrics, Evolve 512) through a Zeiss Alpha Plan/Apo ×100/1.46 oil DIC M27 objective lens. Z-stacks were obtained at focus intervals of 0.4 μm. FiJi/ImageJ software was used to measure the distances between the foci and the periphery.
For the imaging of cells expressing CFP-Mmi1, the following setup was used: confocal microscopy was performed at the Core Facility Bioimaging of the Biomedical Center (LMU Munich) with an inverted Leica SP8 X WLL microscope, equipped with 405-nm laser, WLL2 laser (470–670 nm), and acusto-optical beam splitter. Images were acquired with a HC PL APO ×93/1.30 GLYC motCORR-STED WHITE objective, and Z-stacks were obtained at focus intervals of 0.25 μm. Images were deconvolved using the SVI Huygens suite and FiJi/ ImageJ software was used to measure the distances between the foci and the periphery.
For the imaging of cells expressing CFP-Mmi1, the following setup was used: confocal microscopy was performed at the Core Facility Bioimaging of the Biomedical Center (LMU Munich) with an inverted Leica SP8 X WLL microscope, equipped with 405-nm laser, WLL2 laser (470–670 nm), and acusto-optical beam splitter. Images were acquired with a HC PL APO ×93/1.30 GLYC motCORR-STED WHITE objective, and Z-stacks were obtained at focus intervals of 0.25 μm. Images were deconvolved using the SVI Huygens suite and FiJi/ ImageJ software was used to measure the distances between the foci and the periphery.
6
Live Cell Imaging of Cellular Foci
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Live cell imaging was essentially performed as described (Barrales et al., 2016) . In brief, cells were grown overnight on rich medium to logarithmic phase. Prior to imaging, cells were attached with lectin (Sigma) to glass bottom dishes with a micro well (MatTek). Cells were imaged on a Zeiss Axi-oObserver Z1 confocal spinning disk microscope with an EMM-CCD camera (Photometrics, Evolve 512) through a Zeiss Alpha Plan/Apo 100x/1.46 oil DIC M27 objective lens. Z-stacks were obtained at focus intervals of 0.4 μm. FiJi/ ImageJ software was used to measure the distances between the foci and the periphery.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!