Vale, Ronald D.; Spudich, James A.; Griffis, Eric R. (2021). CIL:11987, Drosophila melanogaster, epithelial cell. In Cell Image Library. UC San Diego Library Digital Collections. Dataset. https://doi.org/10.6075/J0P55M89

TIRF time lapse of myosin-GFP (left), mCherry-tubulin (middle), and overlay (myosin-GFP in green and mCherry-tubulin in red) in wild-type S2 cells from metaphase to anaphase. This cell has many astral microtubules touching the cortex (within the TIRF illumination field) in the bottom left and very few in the top right. Despite this asymmetry (perhaps due to a tilting of the spindle), myosin localization occurs in a normal, symmetrical manner, and myosin clears from both poles.
Video corresponds to video 4 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 50-200 ms 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; mCherryFP
Processing history: unprocessed raw data; color combine
Data qualification: Raw

• Cell Image Library Group ID: 3733

• Drosophila melanogaster

• Epithelial cell
• Myosin regulatory light chain
• Spindle

• Mitotic anaphase
• Mitotic metaphase
• S2

• data
• image
• video

• No linguistic content; Not applicable

Samplenumber: 11987

Doi: https://doi.org/10.6075/J0P55M89

Source data

• Source Record in the Cell Image Library: https://doi.org/10.7295/W9CIL11987

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