Reporter Mouse Lines for Fluorescence Imaging

Abe, Takaya; Fujimori, Toshihiko

doi:10.1111/dgd.12062

Cited by 107 publications

(81 citation statements)

References 120 publications

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“…There are several transgenic mouse lines available which express reporter genes (http://www.findmice.org; Abe and Fujimori, 2013) and their number continues to increase. In this work, we tested three fluorescent transgenic mouse lines and show that this genotyping technique is feasible whether the fluorescent protein gene is randomly inserted or is integrated by homologous recombination into the common ROSA26 locus.…”

Section: Discussionmentioning

confidence: 99%

Non-invasive instant genotyping of fluorescently labeled transgenic mice

Fink

Yau

Kolbe

et al. 2015

ALTEX

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SummaryFluorescence proteins have been useful as genetic reporters for a wide range of applications in biomedical research and are frequently used for the analysis of transgene activity. Here, we show that expression levels of the ubiquitously expressed fluorescent proteins eGFP, mCherry and tdTomato can be measured in transgenic mouse lines with random or targeted integrations. We identified the tail of the mouse as the tissue best suited for quantifying fluorescence intensity and show that expression levels in the tail correlate with gene dose. This allows for instant non-invasive determination of the genetic condition at the transgenic locus (hemizygous/ heterozygous and homozygous) while simultaneously providing an objective comparison for transgene expression levels among different mouse lines. In summary, we demonstrate for the first time that the gene dose of a ubiquitously expressed fluorescence reporter can be reliably quantified and directly linked to the genotype of transgenic mice. Based on this information, animals with the appropriate genotype can be instantly selected without laborious analysis for establishing and breeding of new transgenic lines, reducing the number of "waste" animals. Furthermore, no tissue sampling is necessary, which is a significant refinement of genotyping procedures. Both aspects are important improvements for the genotyping of transgenic mice that follow the principles of the 3Rs (reduction and refinement).

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Section: Discussionmentioning

confidence: 99%

Non-invasive instant genotyping of fluorescently labeled transgenic mice

Fink

Yau

Kolbe

et al. 2015

ALTEX

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“…Fig. 16(b) shows a cross-section of excised mouse colon epithelial tissue; this tissue sample was from a tdTomato mouse [28] Tissue was excised from the colon, rinsed with phosphate-buffered saline (PBS), stained with fluorescein, and mounted on a glass slide for imaging. Features of the colon epithelium including surface profile and colon crypts can be distinguished.…”

Section: Resultsmentioning

confidence: 99%

Multi-Photon Vertical Cross-Sectional Imaging With a Dynamically-Balanced Thin-Film PZT z-Axis Microactuator

Choi

Duan

et al. 2017

J. Microelectromech. Syst.

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Use of a thin-film piezoelectric microactuator for axial scanning during multi-photon vertical cross-sectional imaging is described. The actuator uses thin-film lead-zirconate-titanate (PZT) to generate upward displacement of a central mirror platform, micro-machined from a silicon-on-insulator (SOI) wafer to dimensions compatible with endoscopic imaging instruments. Device modeling in this paper focuses on existence of frequencies near device resonance producing vertical motion with minimal off-axis tilt even in the presence of multiple vibration modes and non-uniformity in fabrication outcomes. Operation near rear resonance permits large stroke lengths at low voltages relative to other vertical microactuators. Highly uniform vertical motion of the mirror platform is a key requirement for vertical cross-sectional imaging in the remote scan architecture being used for multi-photon instrument prototyping. The stage is installed in a benchtop testbed in combination with an electrostatic mirror that performs in-plane scanning. Vertical sectional images are acquired from 15 μm diameter beads and excised mouse colon tissue.

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“…It is not possible, however, to perform microscopy in vivo, and use of histology in investigating wound healing, a dynamic and constantly changing phenomena, is limited. 13 Using K14-Cre/ROSA mT/mG mice with visualization of wounded sites in fluorescent light can increase the precision, by which wound healing can be assessed in mice in vivo on a macroscopic level. It is interesting to note that the wound healing time as determined by macroscopic inspection in normal light was estimated to be *2 days longer than the wound closure time determined using fluorescent light, which suggests that evaluation of wounds in normal light may lead to overestimations of wound closure times in other situations.…”

Section: Discussionmentioning

confidence: 99%

An Improved Humanized Mouse Model for Excisional Wound Healing Using Double Transgenic Mice

Cheng

Borrelli

et al. 2018

Advances in Wound Care

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Objective: Splinting full-thickness cutaneous wounds in mice has allowed for a humanized model of wound healing. Delineating the epithelial edge and assessing time to closure of these healing wounds via macroscopic visualization have remained a challenge. Approach: Double transgenic mice were created by crossbreeding K14-Cre and ROSA mT/mG reporter mice. Full-thickness excisional wounds were created in K14-Cre/ROSA mT/mG mice (n = 5) and imaged using both normal and fluorescent light on the day of surgery, and every other postoperative day (POD) until wound healing was complete. Ten blinded observers analyzed a series of images from a single representative healing wound, taken using normal or fluorescent light, to decide the POD when healing was complete. K14-Cre/ ROSA mT/mG mice (n = 4) were subsequently sacrificed at the four potential days of rated wound closure to accurately determine the histological point of wound closure using microscopic fluorescence imaging. Results: Average time to wound closure was rated significantly longer in the wound series images taken using normal light, compared with fluorescent light (mean POD 13.6 vs. 11.6, *p = 0.008). Fluorescence imaging of histological samples indicated that reepithelialization was complete at 12 days postwounding. Innovation: We describe a novel technique, using double transgenic mice K14-Cre/ROSA mT/mG and fluorescence imaging, to more accurately determine the healing time of wounds in mice upon macroscopic evaluation. Conclusion: The accuracy by which wound healing can be macroscopically determined in vivo in mouse models of wound healing is significantly enhanced using K14-Cre/ROSA mT/mG double transgenic mice and fluorescence imaging.

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Reporter Mouse Lines for Fluorescence Imaging

Cited by 107 publications

References 120 publications

Non-invasive instant genotyping of fluorescently labeled transgenic mice

Non-invasive instant genotyping of fluorescently labeled transgenic mice

Multi-Photon Vertical Cross-Sectional Imaging With a Dynamically-Balanced Thin-Film PZT z-Axis Microactuator

An Improved Humanized Mouse Model for Excisional Wound Healing Using Double Transgenic Mice

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