Topic
Interference reflection microscopy
About: Interference reflection microscopy is a research topic. Over the lifetime, 174 publications have been published within this topic receiving 8635 citations.
Papers published on a yearly basis
Papers
TL;DR: Evidence is presented that the mechanism of cell adhesion does not involve calcium atoms binding cells to substrate by combining with carboxyl groups on cell surface, substrate, and with a cement substance, and that there is no extracellular material between cell and glass in the adhesions.
Abstract: An optical technique for measuring the thickness of thin films has been adapted and evaluated for studying the structure of the adhesion of cells to glass in tissue culture. This technique, which is termed interference reflection microscopy, has been used to study embryonic chick heart fibroblasts. These findings have been observed: in normal culture medium the closest approach of the cell surface to substrate in its adhesions is ca . 100 A, much of the cell surface lying farther away; chemical treatments which bring the cell surface to near its charge reversal point reduce the closest approach of adhesions to ca . 100 A; high osmotic concentration of a non-polar substance, i.e . sucrose, does not affect the distance between cell and substrate in the adhesions. In addition, optical evidence indicates that there is no extracellular material between cell and glass in the adhesions. When cells de-adhere from glass, they appear not to leave fragments behind. The adhesive sites in these fibroblasts appear to be confined to the edge of the side of the cell facing the substrate and to the pseudopods. The significance of this is discussed in relation to the phenomenon of contact inhibition. Evidence is presented that the mechanism of cell adhesion does not involve calcium atoms binding cells to substrate by combining with carboxyl groups on cell surface, substrate, and with a cement substance. Osmium tetroxide fixation results in a final separation of 100 to 200 A between cell and substrate: there are reasons for thinking that this fairly close approach to the condition in life is produced as an artefact. The results can be accounted for only in terms of the action of electrostatic repulsive forces and an attractive force, probably the van der Waals—London forces. Biological arguments suggest that these results are equally applicable for cell-to-cell adhesions.
611 citations
TL;DR: From the secretion of neurotransmitters via synaptic vesicles to the expulsion of cellular waste via contractile vacuoles, exocyTosis and its sequel, endocytosis, are being explored with a variety of new optical tools.
495 citations
TL;DR: Benefits of using TIRF include the ability to obtain high-contrast images of fluorophores near the plasma membrane, very low background from the bulk of the cell, reduced cellular photodamage and rapid exposure times.
Abstract: Total internal reflection fluorescence (TIRF) microscopy can be used in a wide range of cell biological applications, and is particularly well suited to analysis of the localization and dynamics of molecules and events near the plasma membrane. The TIRF excitation field decreases exponentially with distance from the cover slip on which cells are grown. This means that fluorophores close to the cover slip (e.g. within ~100 nm) are selectively illuminated, highlighting events that occur within this region. The advantages of using TIRF include the ability to obtain high-contrast images of fluorophores near the plasma membrane, very low background from the bulk of the cell, reduced cellular photodamage and rapid exposure times. In this Commentary, we discuss the applications of TIRF to the study of cell biology, the physical basis of TIRF, experimental setup and troubleshooting.
398 citations
TL;DR: It is shown that after transformation of fibroblasts by Rous sarcoma virus a majority of the cells have many fewer focal adhesion plaques and now exhibit a cluster of small patches that are immunolabelled for both vinculin and alpha-actinin.
Abstract: It was recently shown by combined immunofluorescence and interference reflection microscopy that a protein named vinculin, along with alpha-actinin, is concentrated at focal adhesion plaques inside cultured normal fibroblasts [Geiger, B. (1979) Cell 18, 193-205]. These plaques are the discrete, isolated sites of strong adhesions formed between the ventral surfaces of the cells and the substrata on which they are grown. We show that after transformation of fibroblasts by Rous sarcoma virus a majority of the cells have many fewer focal adhesion plaques and now exhibit a cluster of small patches that are immunolabelled for both vinculin and alpha-actinin. Such a cluster (rosette) is located near the ventral surface of the cell, usually partly under the nucleus. The significance that these altered distributions of vinculin and alpha-actinin may have for the rounding up and loss of adherence of transformed cells is discussed.
304 citations
TL;DR: This unit will briefly review the history, optical theory, and different hardware configurations used in TIRFM, and provide experimental details and methodological considerations for studying receptors at the plasma membrane in neurons.
Abstract: Total internal reflection fluorescence (TIRF) microscopy (TIRFM) is an elegant optical technique that provides for the excitation of fluorophores in an extremely thin axial region ("optical section"). The method is based on the principle that when excitation light is totally internally reflected in a transparent solid (e.g., coverglass) at its interface with liquid, an electromagnetic field, called the evanescent wave, is generated in the liquid at the solid-liquid interface and is the same frequency as the excitation light. Since the intensity of the evanescent wave exponentially decays with distance from the surface of the solid, only fluorescent molecules within a few hundred nanometers of the solid are efficiently excited. This unit will briefly review the history, optical theory, and different hardware configurations used in TIRFM. In addition, it will provide experimental details and methodological considerations for studying receptors at the plasma membrane in neurons.
238 citations
Related Topics (5)
Performance Metrics
| Year | Papers |
|---|---|
| 2021 | 2 |
| 2020 | 4 |
| 2019 | 2 |
| 2018 | 6 |
| 2017 | 2 |
| 2016 | 6 |