Cell unroofing is any of various methods to isolate and expose the

cell membrane The cell membrane (also known as the plasma membrane (PM) or cytoplasmic membrane, and historically referred to as the plasmalemma) is a biological membrane that separates and protects the interior of all cells from the outside environment ( ...

cell Cell most often refers to: * Cell (biology), the functional basic unit of life Cell may also refer to: Locations * Monastic cell, a small room, hut, or cave in which a religious recluse lives, alternatively the small precursor of a monastery ...

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

cell lysate Lysis ( ) is the breaking down of the membrane of a cell, often by viral, enzymic, or osmotic (that is, "lytic" ) mechanisms that compromise its integrity. A fluid containing the contents of lysed cells is called a ''lysate''. In molecular bio ...

), in ''cell unroofing'' the aim is to tear and preserve patches of the plasma membrane in order to perform ''in situ'' experiments using (microscopy and

biomedical spectroscopy Biomedical spectroscopy is a multidisciplinary research field involving spectroscopic tools for applications in the field of biomedical science. Vibrational spectroscopy such as Raman or infrared spectroscopy is used to determine the chemical c ...

History

The first observation the bi-layer cell membrane was made in 1959 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 by M.V. Nermut. Professor John Heuser made substantial contributions in the field, imaging the detailed internal structure of the membrane and the

cytoskeleton The cytoskeleton is a complex, dynamic network of interlinking protein filaments present in the cytoplasm of all cells, including those of bacteria and archaea. In eukaryotes, it extends from the cell nucleus to the cell membrane and is compos ...

bound to it with extensive use of the electron microscope. It was only after the development of the

atomic force microscope Atomic force microscopy (AFM) or scanning force microscopy (SFM) is a very-high-resolution type of scanning probe microscopy (SPM), with demonstrated resolution on the order of fractions of a nanometer, more than 1000 times better than the diffr ...

operated in liquid that it was possible to image the cell membranes in ''almost''-physiological conditions and to test its mechanical properties.

Methods

* Freeze-fracturing of monolayers *Quick-freeze deep-etch

electron microscopy An electron microscope is a microscope that uses a beam of accelerated electrons as a source of illumination. As the wavelength of an electron can be up to 100,000 times shorter than that of visible light photons, electron microscopes have a hi ...

and

cryofixation Cryofixation is a technique for fixation or stabilisation of biological materials as the first step in specimen preparation for electron microscopy and cryo-electron microscopy. Typical specimens for cryofixation include small samples of plant or an ...

Sonication A sonicator at the Weizmann Institute of Science during sonicationSonication is the act of applying sound energy to agitate particles in a sample, for various purposes such as the extraction of multiple compounds from plants, microalgae and seawe ...

for

atomic force microscopy Atomic force microscopy (AFM) or scanning force microscopy (SFM) is a very-high-resolution type of scanning probe microscopy (SPM), with demonstrated resolution on the order of fractions of a nanometer, more than 1000 times better than the op ...

* Single-cell unroofing

References

{{Reflist, 2 Cell biology Scientific techniques

History

Methods

See also

References