Full-length BRD1, BRD2, BRD13, and SWI3C cDNAs were cloned into the pDONR201 vector and verified by sequencing. Corresponding entry clones were used in LR recombination reactions to transfer the DNA fragments into the gateway-compatible expression vectors pYFN43 and pYFC43 for BiFC assays (Belda-Palazón et al., 2012 (link)) or the pGWB series vector pGWB605 (Nakamura et al., 2010 (link)) for localization analyses. The obtained binary constructs were used to transform Agrobacterium tumefaciens GV3101. Each pGWB605 construct or the combinations of YFPN and YFPC fusion constructs were co-expressed in 6- to 8-week-old Nicotiana benthamiana leaves after leaf infiltration with GV3101 strains containing the tested construct combinations plus an anti-silencing Agrobacterium strain expressing P19. In addition, an Agrobacterium strain transformed with 35S::H2B-RFP was used to visualize the nuclei. Fluorescence was analyzed 2 days after infiltration using a Nikon D-Eclipse C1 laser scanning confocal microscope.
D eclipse c1 laser scanning confocal microscope
Manufactured by Nikon
Sourced in Japan
The D-Eclipse C1 laser scanning confocal microscope is a high-performance imaging system designed for advanced research applications. It utilizes laser excitation and advanced optics to capture detailed, high-resolution images. The core function of the D-Eclipse C1 is to provide researchers with a powerful tool for examining specimens at the cellular and sub-cellular level.
Automatically generated - may contain errors
Lab products found in correlation
5 protocols using d eclipse c1 laser scanning confocal microscope
1
Bait-and-Prey Protein Interactions
2
Quantifying Fluorescence Intensity in Confocal Microscopy
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Images were captured with a D-Eclipse C1 laser scanning confocal microscope (Nikon) with a 40×/1.3 NA oil immersion objective using a 488 nm Ar/Ar–Kr laser line. Laser power and photomultiplier gain were adjusted to minimize background noise rate and saturated pixel and kept constant for all the acquisitions. By using the dedicated module of Fiji software, we measured fluorescence intensity in the designed region of interest of constant area, selecting three representative regions in each image. Quantification was performed by two independent operators.
3
Transient Protein Expression in Nicotiana benthamiana
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Full-length cDNAs corresponding to BDH1 and BDH2 were cloned into the vector pDONR201 and verified by sequencing. Corresponding entry clones were used in LR recombination reactions to transfer the cDNA fragments to the gateway-compatible expression vector pGWB605 [33 (link)]. The binary constructs obtained were used to transform Agrobacterium tumefaciens GV3101. Each pGWB605 construct was coexpressed in 6-week-old Nicotiana benthamiana leaves after leaf infiltration with GV3101 strains containing the tested construct plus an antisilencing agrobacterial strain expressing P19. In addition, the agrobacterial strain transformed with 35S::H2B-RFP was used to visualize the nuclei as described previously [11 (link)]. Fluorescence was analyzed 2 d after infiltration using a Nikon D-Eclipse C1 laser scanning confocal microscope.
4
Cellular Uptake of CG-PEG-CSO-SPIONs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
In vitro cellular uptake of free CG and the CG from the CG-PEG-CSO-SPIONs were investigated by confocal laser scanning microscopy (CLSM) as previously reported [32 (link)] with modifications. In brief, HT-29 cells were cultured in CM and treated with the respective sample as described in Section 2.9.1 . After incubation for 24 h, CSLM cell images were captured using a D-ECLIPSE C1 confocal laser scanning microscope (Nikon, Shinagawa, Japan) at excitation and emission wavelengths of 400 nm and 470 nm, respectively. The extent of cellular uptake was expressed as the fluorescence intensity associated with the CG-PEG-CSO-SPIONs compared to that associated with the free CG solution or blank NPs (PEG-CSO-SPIONs).
5
Neurobiotin Labeling and Morphometric Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Following electrophysiological characterization, a subset of cells was loaded with 0.2% neurobiotin (Vector Laboratories, Burlingame, CA, United States) by passing +100 pA depolarizing rectangular pulses of 50 ms duration at 10 Hz for 5 to 10 min. After loading, slices were left in the recording chamber for a minimum of 5 min to allow neurobiotin diffusion. For neurobiotin visualization sections were incubated overnight at 4°C in Alexa fluor 546-conjugated streptavidin (1:1,000 in blocking solution; Life Technologies, Eugene, OR, United States). After washing in 0.1 M PBS, slices were mounted on slides and coverslipped with Vectashield HardSet Antifade mounting medium. Fluorescent images were taken using a Nikon D-Eclipse C1 confocal laser scanning microscope equipped with a set of lasers covering the 488 and 543 lines using a Nikon achromatic LWD 16×/0.8w objective. Labeled neurons were reconstructed from z-stacks and total dendritic length and dendritic arbor complexity analyzed using the Simple Neurite Tracer Plugin with its built-in Sholl analysis in Fiji (Longair et al., 2011 (link); Schindelin et al., 2012 (link)). Soma size was quantified as soma area measured from z-stack projections. The total number of branches was defined as the sum of the primary, secondary, tertiary, and quaternary processes.
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!