Project
377-Ichinose-RedAlgae
Title
Beta-tubulin dynamics during cell cycles of red algae
Description
Live cell imaging by fluorescence microscopy is a useful tool for elucidating the localization and function of proteins and organelles in single cells. Especially, time-lapse analysis observing the same field sequentially can be used to observe cells of many organisms and analyze the dynamics of intracellular molecules. By single-cell analysis, it is possible to elucidate the characteristics and fluctuations of individual cells, which cannot be elucidated from the data obtained by averaging the characteristics of an ensemble of cells. The primitive red alga Cyanidioschyzon merolae has a very simple structure and is considered a useful model organism for studying the mechanism of organelle division, since the division is performed synchronously with the cell cycle. However, C. merolae does not have a rigid cell wall, and environmental changes such as low temperature or high pH cause morphological change and disruption easily. Therefore, morphological studies of C. merolae typically use fixed cells. In this study, The authors constructed a long-term time-lapse observation system to analyze the dynamics of proteins in living C. merolae cells. From the results, they elucidate the cell division process of single living cells, including the function of intracellular components.
Funding
This study received funds from Grant-in-Aid for Exploratory Research and Grant-Aid for Scientific Research (B).
Title
Time-lapse images of Cyanidioschyzon merolae cells stably expressing beta-tubulin-sfGFP
Description
Time-lapse images of red algae (Cyanidioschyzon merolae) cells stably expressing beta-tubulin-sfGFP. Beta-tubulin change its distribution during a certain period of the cell cycle. After cell division, beta-tubulin-sfGFP is observed in the daughter cells for 90 min and then disappeared during the G1 phase.. Channel1; sfGFP, Channel2 ; auto-fluorescence of chlorophyll, Channel3; bright field
Description
The uracil-auxotrophic mutant (T1) of Cyanidioschyzon merolae stably expressing bata- tubulin-sfGFP. Superfolder GFP (sfGFP), a GFP with high thermal stability is suitable for culture condition of C. merolae at 42 °C.
Cells
red algae (Rhodophyta) (
)
Intrinsic variables
The mutant Cyanidioschyzon merolae, T1, in which the URA5.3 gene is completely deleted
Extrinsic variables
The cells were stabely expressed superfolder GFP (sfGFP), a GFP with high thermal stability
Gene ontology: Biological processes
cell cycle (GO:0007049), cell division (GO:0051301) (
GO:0051301)
Gene ontology: Cellular components
Gene ontology: Molecular functions
Dimensions
1024x1024x1x3x155
Detection methods
wide-field detection
Visualization methods
fluorescent protein tag, autofluorescence
Illumination methods
wide-field detection
Contrast enhancing methods
Resolution enhancing methods
Sample preparation methods
dispersed cells in vitro
Objective
USLSAPO 100XS (OLYMPUS)