Multimodal Imaging of Transgenic Mice

Except adult brain analysis, eGFP and tdTomato were detected by native fluorescence. Images are representative of at least two mice. For brain analysis, serial 60 μm thick coronal vibratome sections were generated and immunolabeled with anti-Tomato dsred as well as anti-Glial Fibrillary Acidic Protein (GFAP) antibodies, performed according to published protocols.^{25 ,36 (link)} Immunolabeled sections were counterstained with Hoechst stain. Atlas images are from the Mouse Brain Library (http://www.mbl.org/mbl_main/atlas.html).³⁷ Fluorescent microscopy of slides was performed using either Zeiss LSM-700 Confocal Microscope Carl Zeiss AG, Oberkochen, Germany) (single plane) and or on a Zeiss Axio Imager M2 with respective ZEN software. Differential interference contrast microscopy of slides was performed using Zeiss Axio Imager M2. Whole-mount brightfield images were performed using an Olympus SZX12 Stereozoom microscope (Olympus Corporation, Tokyo, Japan) or Olympus MVX10 microscope. Whole mount fluorescent images were performed using an Olympus MVX10 microscope excited using an X-Cite Turbo XT600-T fluorescence light source (Excelitas Technologies Corp, Waltham, Massachusetts) at 475 and 575 nm wavelengths. Images were processed using either Canvas X (Canvas GFX), ImageJ, or Photoshop (Adobe Inc., San Jose, California).