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Cell unroofing
Iztwoz: Page created by The Xiao Li
[[File:Sketch cell unroofing.PNG|thumb|right|400px|The most common processes of cell unroofing. (left) Sandwich of two cells between two coverslips. (right) Lateral flux of medium allows to break the cells.]]<!--no px size to allow auto-sizing per user-->
'''Cell unroofing''' is any of various methods to isolate and expose the [[cell membrane]] of [[cell]]s. Differently from the more common membrane '''extraction protocols''' performed with multiple steps of centrifugation (which goal is to separate the membrane fraction from a [[lysis|cell lysate]]), in ''cell unroofing'' the aim is to tear and preserve patches of the plasma membrane in order to perform ''in situ'' experiments like (microscopy and spectroscopy).
==History==
The first observation the bi-layer cell membrane was made in 1959<ref>Liquid error: wrong number of arguments (1 for 2)</ref> on a section of a cell using the electron microscope.
But the first micrograph of the internal side of a cell dates back to 1977<ref>Liquid error: wrong number of arguments (1 for 2)</ref> by M.V. Nermut. Professor [[John Heuser]] made substantial contributions in the field, imaging the detailed internal structure of the membrane and the [[cytoskeleton]] bound to it with extensive use of the electron microscope.
It was only after the development of the [[Atomic force microscopy|atomic force microscope]] operated in liquid that it was possible to image and cell membranes in ''almost''-physiological conditions<ref>Liquid error: wrong number of arguments (1 for 2)</ref> and to test its mechanical properties<ref name=":0" />.
== Methods ==
* Freeze-fracturing of monolayers<ref>Liquid error: wrong number of arguments (1 for 2)</ref>
*Quick-freeze deep-etch [[electron microscopy]]<ref>Liquid error: wrong number of arguments (1 for 2)</ref>
* Sonication for [[Atomic force microscopy]]<ref>Liquid error: wrong number of arguments (1 for 2)</ref>
* Single-cell unroofing <ref name=":0">Liquid error: wrong number of arguments (1 for 2)</ref>
== References ==
'''Cell unroofing''' is any of various methods to isolate and expose the [[cell membrane]] of [[cell]]s. Differently from the more common membrane '''extraction protocols''' performed with multiple steps of centrifugation (which goal is to separate the membrane fraction from a [[lysis|cell lysate]]), in ''cell unroofing'' the aim is to tear and preserve patches of the plasma membrane in order to perform ''in situ'' experiments like (microscopy and spectroscopy).
==History==
The first observation the bi-layer cell membrane was made in 1959<ref>Liquid error: wrong number of arguments (1 for 2)</ref> on a section of a cell using the electron microscope.
But the first micrograph of the internal side of a cell dates back to 1977<ref>Liquid error: wrong number of arguments (1 for 2)</ref> by M.V. Nermut. Professor [[John Heuser]] made substantial contributions in the field, imaging the detailed internal structure of the membrane and the [[cytoskeleton]] bound to it with extensive use of the electron microscope.
It was only after the development of the [[Atomic force microscopy|atomic force microscope]] operated in liquid that it was possible to image and cell membranes in ''almost''-physiological conditions<ref>Liquid error: wrong number of arguments (1 for 2)</ref> and to test its mechanical properties<ref name=":0" />.
== Methods ==
* Freeze-fracturing of monolayers<ref>Liquid error: wrong number of arguments (1 for 2)</ref>
*Quick-freeze deep-etch [[electron microscopy]]<ref>Liquid error: wrong number of arguments (1 for 2)</ref>
* Sonication for [[Atomic force microscopy]]<ref>Liquid error: wrong number of arguments (1 for 2)</ref>
* Single-cell unroofing <ref name=":0">Liquid error: wrong number of arguments (1 for 2)</ref>
== References ==
January 05, 2019 at 04:34PM
