Cryo-electron microscopy (EM) imaging and analysis of images was performed as described earlier (Chadda et al., 2018 (link); Cliff et al., 2020 (link)). Briefly, liposomes were freeze-thawed seven times, and then extruded through a 400 nm nucleopore filter (GE Life Sciences) 21 times. Three µL of the undiluted sample was loaded onto a glow-discharged Lacey carbon support film (Electron Microscope Sciences), blotted, and plunged into liquid ethane using a Vitrobot System (FEI). Images were collected on a FEI Titan Krios G3 300kV Cryo-TEM microscope with a Gatan K2 Summit Direct electron detector (GATAN). Magnifications of 6500x, 33,000x, and 53,000x were used. For size determination, liposomes were manually outlined in Fiji and ImageJ (Schindelin et al., 2012 (link); Schindelin et al., 2015 (link)) to measure the outer radii of all liposomes, including those located on the carbon. Multilamellarity was manually counted as the fraction of vesicles containing more than one bilayer. Liposome size distribution source data is provided in Figure 4—source data 1 .
400 nm nucleopore filter
Manufactured by GE Healthcare
The 400-nm nucleopore filter is a laboratory equipment designed to separate and isolate particles, cells, or molecules based on their size. It features a membrane with uniformly distributed pores measuring 400 nanometers in diameter, allowing for precise filtration and separation of materials. This filter is commonly used in various research and analytical applications that require size-based separation or purification of samples.
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Lab products found in correlation
3 protocols using 400 nm nucleopore filter
1
Cryo-EM Analysis of Liposome Morphology
2
Liposome Morphology Characterization Protocol
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Liposomes were freeze-thawed seven times, incubated at room temperature, and then extruded through a 400-nm nucleopore filter (GE Life Sciences) 21 times before sample freezing. 3 µl of the undiluted sample was loaded onto glow-discharged Lacey carbon support films (Electron Microscope Sciences), blotted, and plunged into liquid ethane using a Vitrobot System (FEI). Images were collected at 300 kV on a JEOL 3200 fs microscope with a K2 Summit direct electron detector camera (GATAN). Magnifications of 15,000 and 30,000 were used. For size determination, liposomes were manually outlined in Fiji and ImageJ (Schindelin et al., 2012 (link); Schneider et al., 2012 (link)) to measure the outer radii of all liposomes, including those located on the carbon. The normalized frequency histograms were averaged from two independent preparations (samples sizes, 140 and 686), and the mean ± SD is reported in Table 1 .
3
Cryo-EM Imaging and Analysis of Liposomes
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Cryo-electron microscopy (EM) imaging and analysis of images was performed as described earlier (Chadda et al., 2018; (link)Cliff et al., 2019) (link). Briefly, liposomes were freeze-thawed seven times, and then extruded through a 400-nm nucleopore filter (GE Life Sciences) 21 times. 3 µL of the undiluted sample was loaded onto a glow-discharged Lacey carbon support film (Electron Microscope Sciences), blotted, and plunged into liquid ethane using a Vitrobot System (FEI). Images were collected on a FEI Titan Krios G3 300kV Cryo-TEM microscope with a Gatan K2 Summit Direct electron detector (GATAN). Magnifications of 6500x, 33 000x and 53 000x were used.
For size determination, liposomes were manually outlined in Fiji and ImageJ (Schindelin et al., 2012 (link)(Schindelin et al., , 2015) ) (link) to measure the outer radii of all liposomes, including those located on the carbon. Multilamellarity
(which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission.
was manually counted as the fraction of vesicles containing more than one bilayer. Liposome size distribution source data is provided in Fig. 4 -source data 1.
For size determination, liposomes were manually outlined in Fiji and ImageJ (Schindelin et al., 2012 (link)(Schindelin et al., , 2015) ) (link) to measure the outer radii of all liposomes, including those located on the carbon. Multilamellarity
(which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission.
was manually counted as the fraction of vesicles containing more than one bilayer. Liposome size distribution source data is provided in Fig. 4 -source data 1.
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