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<div><br></div>I have seen incredibly bright results with tandem dimer-tomato, but have not made a germ-line animal yet. <div><br><div>Animals harboring apparently single tdT (+) neurons were easily detected even with a 2.5 objective. </div><div><br></div><div>We did not find the signal to be too interfering to visualization of neighboring cells when expressed in just a few cells, however, when it was expressed in skin we could not see through tdT very well when using the TR filter. </div><div><br></div><div>Best, </div><div>W. Todd Penberthy </div><div><br><div><div>On Oct 21, 2008, at 11:22 AM, Philippe Herbomel wrote:</div><br class="Apple-interchange-newline"><blockquote type="cite">Dear All,<div><br></div><div>We are wondering about the best red fluorescent reporter to make transgenic zebrafish with.</div><div>If one does not want to target the protein to a particular compartment (membrane or nucleus), DsRed2, the most used presently, is probably the best (very bright, with reasonable maturation time).</div><div>If otherwise, the most used presently is mCherry, which is quite (4-fold) less bright. </div><div>Now from the Tsien lab papers' data, td-Tomato appears both much (over 5-fold) brighter than mCherry, and still compatible with membrane or nuclear targeting; </div><div>and indeed at least two papers then provided evidence that td-tomato was not only far superior to mcherry but even to GFP (in terms of brightness) as a reporter in transgenic mice.</div><div>Now surprisingly, when a colleague of us injected either mcherry or tdtomato mRNA into 1-cell zebrafish embryos,</div><div>she found td-Tomato to be less bright than mcherry.</div><div>So my question is: has anyone found this too, and/or tried to make transgenic zebrafish with td-Tomato as reporter ? And what about its actual brightness (and decay upon illumination) in zebrafish embryos/larvae, as compared to the mcherry and dsred2 ?</div><div><br></div><div>Thanks in advance for your testimonies on this. </div><div>I will post a summary of your responses !</div><div><br></div><div>Philippe Herbomel </div><div><div><br></div><div><br></div></div><div> <span class="Apple-style-span" style="border-collapse: separate; color: rgb(0, 0, 0); font-family: Helvetica; font-size: 12px; font-style: normal; font-variant: normal; font-weight: normal; letter-spacing: normal; line-height: normal; orphans: 2; text-align: auto; text-indent: 0px; text-transform: none; white-space: normal; widows: 2; word-spacing: 0px; -webkit-border-horizontal-spacing: 0px; -webkit-border-vertical-spacing: 0px; -webkit-text-decorations-in-effect: none; -webkit-text-size-adjust: auto; -webkit-text-stroke-width: 0; "><span class="Apple-style-span" style="border-collapse: separate; color: rgb(0, 0, 0); font-family: Helvetica; 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