We used Chlamydomonas reinhardtii mat3-4 cells (strain CC-3994)87 (link), which exhibit superior vitrification due to their small size. The strain was acquired from the Chlamydomonas Resource Center, University of Minnesota, St. Paul. Cells were grown until the mid-log phase in Tris-acetate-phosphate (TAP) medium under constant light exposure and bubbling with a normal atmosphere. Vitrification and FIB milling were performed as previously described88 (link),89 . Using a Vitrobot Mark 4 (FEI), cells in suspension (4 μl of ∼1000 cells per µl) were blotted onto R2/1 carbon-coated 200-mesh copper grids (Quantifoil Micro Tools) and plunge frozen in a liquid ethane/propane mixture. Grids were then mounted into Autogrid supports (FEI) and transferred into either a FEI Scios or FEI Quanta dual-beam FIB/SEM instrument. The grids were coated with an organometallic platinum layer by the gas injection system (FEI), and cells were thinned from both sides with a gallium ion beam to a final thickness of ∼100–200 nm.
R2 1 carbon coated 200 mesh copper grids
Manufactured by Quantifoil
The R2/1 carbon-coated 200-mesh copper grids are a type of specimen support designed for transmission electron microscopy (TEM) applications. They consist of a copper grid with a mesh size of 200 and a carbon coating. The primary function of these grids is to provide a stable and conductive support for samples during TEM analysis.
Automatically generated - may contain errors
2 protocols using r2 1 carbon coated 200 mesh copper grids
1
Vitrification and FIB Milling of Chlamydomonas Cells
2
Chlamydomonas Cells Vitrification and FIB Milling
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For FIB-milling, Chlamydomonas reinhardtii mat3-4cells (strain CC-3994) (Umen and Goodenough, 2001) , which exhibit superior vitrification due to their small size, were used. They were acquired from the Chlamydomonas Resource Center, University of Minnesota, St. Paul. Cells were grown until midlog phase in Tris-acetate-phosphate (TAP) medium under constant light exposure and bubbling with normal atmosphere. Vitrification and FIB sample preparation were performed as previously described (Schaffer et al., 2015 (Schaffer et al., , 2017)) . Using a Vitrobot Mark 4 (FEI), cells in suspension (4 μl of ∼1,000 cells per µl) were blotted onto R2/1 carbon-coated 200-mesh copper grids (Quantifoil Micro Tools) and plunge frozen in a liquid ethane/propane mixture. Grids were then mounted into Autogrid supports (FEI) and transferred into either a FEI Scios or FEI Quanta dual-beam FIB/SEM instrument. The grids were coated with an organometallic platinum layer by the gas injection system (FEI), and cells were thinned from both sides with a gallium ion beam to a final thickness of ∼100-200 nm.
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!