For visualizing the macerated single cells, two drops of Calcoflour White (1 g L−1, Sigma-Aldrich) were put on a glass slide, on top of the macerated single cells. A LEICA SP5 was equipped with a 60 ×/0.9 water objective, and a diode laser (λexc = 405 nm). The stained cellulose was detected at 425–475 nm. Images were obtained with a step size of 0.4 μm, covering the entire volume of cells (50–100 planes) via the Leica software (LAS AF 3.1). The software Imaris (8.4.0, Bitplane) was used for 3D reconstruction of the cells.
Calcoflour white
Manufactured by Merck Group
Sourced in Germany
Calcofluor white is a fluorescent dye commonly used in microscopy to stain cellulose and chitin in biological samples. It binds to these polysaccharides, allowing their visualization under ultraviolet or blue light illumination.
Automatically generated - may contain errors
4 protocols using calcoflour white
1
3D Visualization of Macerated Cells
2
Mitochondrial Staining and Imaging
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Log phase cell cultures were exposed to different
drugs at 30 °C, for times indicated in figure legends. Cells
were isolated from a log phase culture and warmed to 37 °C. Mitotracker
Green FM (c.n.M7514 ThermoFisher) was added to a final concentration
of 1 mM. Cells were incubated in the dark for 0–120 min. Cells
were then washed with 1× PBS buffer and resuspended in SDC medium
(0.17% yeast nitrogen base with ammonium sulfate [Formedium], 2% glucose)
supplemented with amino and nucleic acids)600 containing 1 mg/mL calcoflour white (by Sigma-Aldrich) for imaging
each time point. DAPI (4′,6-diamidino-2-phenylindole, Sigma-Aldrich)
was suspended in double distilled water and added to cells at 10 μg/mL
final concentration in PBS with cells at 25 °C for 5 min.
drugs at 30 °C, for times indicated in figure legends. Cells
were isolated from a log phase culture and warmed to 37 °C. Mitotracker
Green FM (c.n.M7514 ThermoFisher) was added to a final concentration
of 1 mM. Cells were incubated in the dark for 0–120 min. Cells
were then washed with 1× PBS buffer and resuspended in SDC medium
(0.17% yeast nitrogen base with ammonium sulfate [Formedium], 2% glucose)
supplemented with amino and nucleic acids)600 containing 1 mg/mL calcoflour white (by Sigma-Aldrich) for imaging
each time point. DAPI (4′,6-diamidino-2-phenylindole, Sigma-Aldrich)
was suspended in double distilled water and added to cells at 10 μg/mL
final concentration in PBS with cells at 25 °C for 5 min.
3
Visualizing Cellulose and Callose in Plant Roots
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To stain cellulose, Mitra and Loque [61 ] protocol for Calcofluor staining was followed with some modifications. Briefly, the roots were transferred to 2 ml tubes and stained with 0.02% calcoflour white (Sigma) for 5 min. The primary root was placed on the microscope glass slide and transverse section of middle part of the roots was prepared using sharp blade. Calcofluor White was visualized using an epifluorescence microscope (Zeiss Imager M2).
The staining of callose was done according to Muller et al. [62 (link)] with some modifications. Briefly, the roots were stained with 0.1% (w/v) aniline blue solution in 0.1 M sodium phosphate buffer (pH = 7.2) for 1.5 h. The cross-section of roots was done as described above and visualized under epifluorescence microscope (Zeiss Imager M2).
The staining of callose was done according to Muller et al. [62 (link)] with some modifications. Briefly, the roots were stained with 0.1% (w/v) aniline blue solution in 0.1 M sodium phosphate buffer (pH = 7.2) for 1.5 h. The cross-section of roots was done as described above and visualized under epifluorescence microscope (Zeiss Imager M2).
4
Microwave-assisted Microscopy of Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cross-sections (2 mm thick) from the central region of 3/4 DAF were used for microwave-assisted aldehyde fixation, substitution and embedding as described [30 (link)]. Semi- and ultra-microtomy, as well as histological and ultrastructural analyses, were performed as previously described. [128 (link)]. Cell wall staining was carried out with Calcoflour white (Sigma-Aldrich, Taufkirchen, Germany) 1 mg/mL in 50 mM phosphatebuffer, pH 7.2. Nuclei were stained with 1 mg/mL DAPI solution. Fluorescence was recorded in a LSM780 confocal laser scanning microscope (Carl Zeiss, Jena, Germany) using a 405 nm laser for excitation in combination with a 410–500 nm bandpass for detection.
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!