Tube pulling tension measurements were performed using a modified version of published setup15 (link), which allows simultaneous bright-field and FLIM imaging on an inverted Nikon Eclipse Ti A1R microscope. A membrane nanotube was formed by pulling away a micropipette-aspirated GUV, whose membrane was attached to a streptavidin-coated bead (3.05 mm diameter, Spherotec, Lake Forest, Illinois, USA) held in a fixed optical trap. The trap was obtained by focusing an ytterbium fiber laser (IPG laser, Burbach, Germany) through a 100X 1.3 NA oil immersion objective (Nikon, Tokyo, Japan). Force measurements were made measuring the displacement of the bead in the optical trap via a C-MOS Camera (Pixelink, Ottawa, Canada) with a home-made video recorder and bead tracking software under Matlab. The force F exerted on the bead was calculated from the Hooke’s law: F = k.Δx, where k is the stiffness of the trap (k = 8.58 pN.pix-1. W-1) and Δx the displacement of the bead from its initial, zero force position.
Protocol detail
Membrane Nanotube Force Measurements
Partial Protocol Preview
This section provides a glimpse into the protocol.
The remaining content is hidden due to licensing restrictions, but the full text is available at the following link:
Access Free Full Text.
Corresponding Organization : University of Geneva
Other organizations : MRC Laboratory of Molecular Biology
Top 5 similar protocols
Variable analysis
independent variables
- Pulling away a micropipette-aspirated GUV to form a membrane nanotube
- Force exerted on the bead, calculated from the Hooke's law: F = k.Δx, where k is the stiffness of the trap (k = 8.58 pN.pix^-1.W^-1) and Δx the displacement of the bead from its initial, zero force position
- Bead diameter (3.05 mm)
- Trap stiffness (k = 8.58 pN.pix^-1.W^-1)
- Microscope setup (Inverted Nikon Eclipse Ti A1R microscope)
- Laser source (Ytterbium fiber laser, IPG laser, Burbach, Germany)
- Objective (100X 1.3 NA oil immersion, Nikon, Tokyo, Japan)
- Camera (C-MOS Camera, Pixelink, Ottawa, Canada)
- Positive control: Published setup15 (link)
- Negative control: Not mentioned
Annotations
Based on most similar protocols
Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.
Sign up for free or login to display all annotations
As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.
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!