Cells were fixed in 4% paraformaldehyde for 15 minutes and blocked with 5% goat serum in PBS for 1 hour at room temperature. Cells were then incubated with mouse anti-α-sarcomeric actinin monoclonal antibody (1:500, Sigma-Aldrich) diluted in 1% goat serum in PBS overnight at 4°C. The next day, cells were washed with PBS followed by staining with Alexa-594 conjugated goat-anti-mouse secondary antibody (1:200, Invitrogen) and Alexafluo 488 phalloidin (1:200, Invitrogen). Counterstaining was performed with Vectashied containing DAPI (Vector Labs). Images were captured with a Nikon A1R confocal microscope using a Nikon CFI Plan Apo VC 60X water immersion objective. For each condition, at least 4 biological specimens were prepared and imaged, with at least 4 images taken per specimen.
Cfi plan apo vc 60x
Manufactured by Nikon
The CFI Plan Apo VC 60X is a high-performance objective lens for optical microscopy. It is designed to provide clear, high-resolution images with a wide field of view. The lens features a numerical aperture of 1.40 and a working distance of 0.17 mm, making it suitable for a variety of microscopy applications.
Automatically generated - may contain errors
Lab products found in correlation
Alexa 594 conjugated goat anti mouse secondary antibody, by Thermo Fisher Scientific (1 mentions)
A1r confocal microscope, by Nikon (1 mentions)
Oil immersion objective, by Nikon (1 mentions)
Hit stage top incubator, by Tokai Hit (1 mentions)
Ff495 di03, by IDEX Corporation (1 mentions)
Orca ag cooled ccd camera, by Hamamatsu Photonics (1 mentions)
Ff01 520 35, by IDEX Corporation (1 mentions)
Te2000, by Nikon (1 mentions)
4 protocols using cfi plan apo vc 60x
1
Immunocytochemical Analysis of Sarcomeric Actin
2
Imaging Axon Dynamics in Zebrafish
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For live imaging, embryos were anesthetized in 0.02% tricaine and mounted in 1% low melting agarose in 10 mM HEPES E3 medium as previously described (Andersen et al., 2011 (link)). Live high speed imaging of EB3-GFP comets was performed with an Opterra Swept-Field confocal microscope (Bruker Nano Surfaces FM) equipped with a Nikon CFI Plan Apo VC 60x (NA 1.40) or 100x oil-immersion objective (NA 1.40). Embryos were imaged at stages between 18 and 26 hpf, while peripheral axons are initiating and arborizing. Z-stacks of 5–50 1-μm optical sections were captured at 2–9 s time intervals, for total durations between 3 and 20 min.
3
Measuring Actin Dynamics in Cardiomyocytes
4
Cytosolic pH Measurement in HeLa Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The cytosolic pH of HeLa cells was measured with BCECF (2′-7′-bis(carboxyethyl)-5(6)-carboxyfluorescein) as the following protocol. The cells were treated with 10 μM BCECF acetoxymethyl ester (Molecular probes) in Hank’s balanced salt solution (HBSS) for 15 min at 37 °C, followed by washing with HBSS twice. Fluorescence intensity of BCECF of the cells (n = 6) was measured at 37 °C in DMEM using a Nikon TE2000 (Nikon Instruments) equipped a CFI Plan Apo VC 60x oil-immersion objective lens, 1.40 numerical aperture (Nikon Instruments). The following filters (Semrock) were used for the measurement; excitation filters (FF01-445/20 and FF01-482/18), an emission filter (FF01-520/35), and a dichroic mirror (FF495-Di03). Images were captured with an ORCA-AG cooled CCD camera (Hamamatsu photonics). Calibration curve was obtained by imaging BCECF-loaded cells in pH-controlled buffers containing 150 mM KCl, 20 mM NaCl, 0.5 mM MgCl2, 0.5 mM CaCl2, 10 mM MES, 10 mM HEPES, 10 μM monensin and 10 μM nigericin.
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!