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Author: Xia, J
Author: Truant, R
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Methods Article

Practical three color live cell imaging by widefield microscopy

Jianrun Xia1, Song Hon H. Kim1, Susan Macmillan1 and Ray Truant1*

1 Department of Biochemistry and Biomedical Sciences, McMaster University. HSC 4H24A 1200 Main Street West, Hamilton, Ontario, L8N 3Z5. Canada.

* To whom correspondence should be addressed: Ray Truant, Department of Biochemistry and Biomedical Sciences, McMaster University. HSC 4H24A 1200 Main Street West, Hamilton, Ontario, L8N 3Z5. Canada. Phone: 01-905-525-9140 ext. 22450. Fax: 01-905-522-9033. Email: truantr@mcmaster.ca

Biol. Proced. Online 2006;8:63-68. doi:10.1251/bpo119
Submitted: April 10, 2006; Accepted: June 13, 2006; Published: July 21, 2006.

Indexing terms: Green Fluorescent Proteins; Blue Fluorescent Protein, Aequorea Victoria.


Figure 2 Enlarged

Fig. 2:

Live cell Imaging over Time of RGB Fluorescent Proteins That Can form Bright Puncta. A. Three color imaging using a triple cube filter set for DAPI/FITC/Texas Red (Semrock DA/FI/TX-B) and switching only excitation filters for three proteins that form independent bright puncta: huntingtinExon1Q138-mRFP, Mito-eGFP, and PSCFP2-NXf1. Bright puncta in each channels are highlighted with white arrows. B. Time course of mRFP-huntingtin Q138 exon1 fragment, prone to aggregation, eGFP-ASF2, nuclear speckle protein, and PSCFP2-NXF1, a mRNA export factor in STHdh cells. Three color images were captured every minute for 75 minutes. Developing polyglutamine inclusions of huntingtin can be seen in the red channel without bleed to green or blue channels and no artifactual accumulation of the other two fusion proteins into the large polyglutamine aggregates (top row). Scale bar is 10 μm. Also in supplemental video 1.

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