Vale, Ronald D.; Spudich, James A.; Griffis, Eric R. (2021). CIL:11993, Drosophila melanogaster, epithelial cell. In Cell Image Library. UC San Diego Library Digital Collections. Dataset. https://doi.org/10.6075/J0T43RVQ

TIRF time lapse of myosin-GFP in a cell with a symmetric monopolar spindle from metaphase to anaphase (cell was depleted of Klp61F [kinesin-5] and BubR1 by RNAi). At anaphase, myosin-GFP concentrates at the cell perimeter and becomes depleted from the central regions of the adhered cortex. Slowly, myosin breaks symmetry and ultimately relocalizes to the middle. This video corresponds to the image in Fig. 6 and video 10 from J Cell Bio 186:727-738, 2009.
Total internal reflection microscopy was performed using a microscope (Perfect-focus TE2000; Nikon) with a 100×, 1.45 NA objective (Nikon) and illumination from either a 488-nm argon laser (100 mW) or a 491-nm solid-state laser (100 mW) and a 561-nm solid-state laser (50 mW). For dual-color TIRF microscopy, we used a triplepass dichroic filter (z491/561/633rpc) and changed the emission filter (ET525/50 or ET595/50; both from Chroma Technology Corp.) with a filter wheel placed before the camera. In some cases, an excitation notch filter was used in the filter cube (NF01-405/488/561/635; Semrock, Inc.). Images were typically captured every 2-3 s with a 50-200ms exposure with an EM charge-coupled device camera (iXon; Andor Technology). The microscope was controlled and images were acquired using open source MicroManager software (http://www.micro-manager.org).

• 2021

• Griffis, Eric R.
• Spudich, James A.
• Vale, Ronald D.

Preparation: living tissue
Relation to intact cell: dispersed cells in vitro
Item type: recorded image
Imaging mode: TIRF
Parameter imaged: fluorescence emission
Source of contrast: distribution of a specific protein
Visualization methods: EGFP
Processing history: unprocessed raw data
Data qualification: Raw;spatialmeasurements

• Cell Image Library Group ID: 3733

• Drosophila melanogaster

• Epithelial cell
• Myosin regulatory light chain

• BubR1 knockdown
• Kinesin 5 knockdown
• Mitotic anaphase
• Mitotic metaphase
• S2

• data
• image
• video

• No linguistic content; Not applicable

Samplenumber: 11993

Doi: https://doi.org/10.6075/J0T43RVQ

Source data

• Source Record in the Cell Image Library: https://doi.org/10.7295/W9CIL11993

Reference

• J Cell Bio 186:727-738, 2009 https://www.ncbi.nlm.nih.gov/pubmed/?term=19720876
• Vale RD, Spudich JA, Griffis ER. Dynamics of myosin, microtubules, and Kinesin-6 at the cortex during cytokinesis in Drosophila S2 cells. J Cell Biol. 2009 Sep 7;186(5):727-38. https://doi.org/10.1083/jcb.200902083

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2022-08-12