For microscopy, cell cultures diluted in water or phosphate-buffered saline (PBS) solution, pH 7.0, and mixed with an equal volume of 0.01% (w/v) solution of one or more dyes (all from Sigma Aldrich, except calcofluor white from Megazyme). Cells were imaged with a 40× or 60× objectives on a laser scanning confocal microscope (Carl Zeiss, LSM 700), beam splitter (MBS 405/488/555/639), and multiple laser/filter combinations. Separate acquisition tracks with the following excitation and emission wavelengths were used to acquire Venus fluorescence (488 nm and BP 450–550), calcofluor (405 nm and BP 420–550), TB, propidium iodide, and congo red (639 nm and LP 640). Images were acquired using the ZEN 2011 (black edition) from Carl Zeiss and then processed uniformly in ImageJ [47 (link)]. To quantify cell numbers and sizes, the microscopy images were segmented using the web version (http://yeastspotter.csb.utoronto.ca ) of the YeastSpotter tool [48 (link)], and particles were then measured in ImageJ with the Analyze Particles (size = 3–40, circularity = 0.80–1.00) command.
Calcofluor white
Manufactured by Megazyme
Calcofluor white is a fluorescent dye used in microscopy to stain and visualize cellulose and chitin structures in a wide range of biological samples, including plants, fungi, and bacteria. It binds to these polysaccharides and emits a bright blue fluorescence when exposed to ultraviolet or blue light.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using calcofluor white
1
Quantitative Microscopy of Cellular Morphology
2
Microscopic Analysis of Cell Cultures
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For microscopy, cell cultures diluted in water or phosphate-buffered saline (PBS) solution, pH 7.0, and mixed with an equal volume of 0.01% (w/v) solution of one or more dyes (all from Sigma Aldrich, except calcofluor white from Megazyme). Cells were imaged with a 40x or 60x objectives on a laser scanning confocal microscope (Carl Zeiss, LSM 700), beam splitter (MBS 405/488/555/639), and multiple laser/filter combinations. Separate acquisition tracks with the following excitation and emission wavelengths were used to acquire Venus fluorescence (488 nm and BP 450-550), calcofluor (405 nm and BP 420-550), TB, propidium iodide, and congo red (639 nm and LP 640). Images were acquired using the ZEN 2011 (black edition) from Carl Zeiss and then processed uniformly in ImageJ [47] . To quantify cell numbers and sizes, the microscopy images were segmented using the web version (http://yeastspotter.csb.utoronto.ca) of the YeastSpotter tool [48] , and particles were then measured in ImageJ with the Analyze Particles (size=3-40, circularity=0.80-1.00) command.
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!