Document Detail


Preparation of an imaging chamber for visualizing single molecules in living dictyostelium cells.
MedLine Citation:
PMID:  22383649     Owner:  NLM     Status:  In-Data-Review    
Abstract/OtherAbstract:
Environmental changes result in signaling events at cell membranes. To develop the means to understand these events at the molecular level, it is essential to become familiar with the stochastic nature of signaling molecules in living cells. Using total internal reflection fluorescent microscopy (TIRFM), these signaling events can be directly observed at the single-molecule level. This protocol describes the preparation of an imaging chamber to visualize single molecules in living Dictyostelium cells, which are highly sensitive to chemoattractant stimulation. It also describes treatment of the cells to allow visualization. Chemotactically competent cells are treated with the desired chemical, and the cell suspension is delivered onto coverslips and then overlaid with a thin agarose sheet in preparation for imaging. The cells are typically treated with a fluorescently labeled stimulant or inhibitor. Alternatively, the cells can be stimulated with photoreactive chemicals by using caged compounds. A caged compound is photoactivatable by irradiation with ultraviolet (UV) light, which cleaves a linker conjugating a caging moiety and a chemical. Thus, at a desired moment during recording of the single-molecule images, the concentration of the chemicals can be increased by photolysis of the caged compounds included in the buffer.
Authors:
Satomi Matsuoka; Yukihiro Miyanaga; Toshio Yanagida; Masahiro Ueda
Related Documents :
9389439 - From flow cytometric brdurd data to cell population growth and doubling time.
20352219 - The turnover of synovial t cells is higher than in t cells in the peripheral blood in p...
7237289 - K+, na+, and mg2+ content and permeability of methanospirillum hungatei and methanobact...
11093149 - Activation-induced t cell death occurs at g1a phase of the cell cycle.
2269869 - Switching at the cellular level in the white-opaque transition of candida albicans.
1648559 - Growth phase-regulated expression of bola and morphology of stationary-phase escherichi...
6279649 - Kinetic aspects of hemoglobin.haptoglobin-receptor interaction in rat liver plasma memb...
3335499 - Isolation and characterization of chinese hamster ovary cell mutants deficient in acyl-...
8704279 - P-glycoprotein expression in ovarian cancer cell line following treatment with cisplatin.
Publication Detail:
Type:  Journal Article     Date:  2012-03-01
Journal Detail:
Title:  Cold Spring Harbor protocols     Volume:  2012     ISSN:  1559-6095     ISO Abbreviation:  Cold Spring Harb Protoc     Publication Date:  2012  
Date Detail:
Created Date:  2012-03-02     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  101524530     Medline TA:  Cold Spring Harb Protoc     Country:  United States    
Other Details:
Languages:  eng     Pagination:  -     Citation Subset:  IM    
Export Citation:
APA/MLA Format     Download EndNote     Download BibTex
MeSH Terms
Descriptor/Qualifier:

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


Previous Document:  Preparation of chemotactically competent dictyostelium cells.
Next Document:  Visualizing Single Molecules in Living Dictyostelium Cells Using Total Internal Reflection Fluoresce...