Journal article

Intravital microscopy of dynamic single-cell behavior in mouse mammary tissue

Caleb A Dawson, Scott N Mueller, Geoffrey J Lindeman, Anne C Rios, Jane E Visvader

Nature Protocols | NATURE RESEARCH | Published : 2021

Abstract

Multiphoton intravital imaging is essential for understanding cellular behavior and function in vivo. The adipose-rich environment of the mammary gland poses a unique challenge to in vivo microscopy due to light scattering that impedes high-resolution imaging. Here we provide a protocol for high-quality, six-color 3D intravital imaging of regions across the entire mouse mammary gland and associated tissues for several hours while maintaining tissue access for microdissection and labeling. An incision at the ventral midline and along the right hind leg creates a skin flap that is then secured to a raised platform skin side down. This allows for fluorescence-guided microdissection of connectiv..

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Grants

Awarded by Australian National Health and Medical Research Council (NHMRC)


Awarded by NHMRC


Funding Acknowledgements

We thank F. Jackling for animal management and S. Devi and R. Yip for imaging assistance. We are grateful to the Walter and Eliza Hall Institute (WEHI) Center for Dynamic Imaging and WEHI Bioservices. This work was supported by the Australian National Health and Medical Research Council (NHMRC) grants #1016701, 1113133; NHMRC IRIISS; the Victorian State Government through VCA funding and Operational Infrastructure Support; and the Australian Cancer Research Foundation. C.A.D was supported by an Australian Government Research Training Program Scholarship; A.C.R. was supported by a National Breast Cancer Foundation (NBCF)/Cure Cancer Australia Fellowship; G.J.L. by NHMRC Fellowship #1078730, #1175960; J.E.V. by NHMRC Fellowships #1037230, 1102742; S.N.M. by NHMRC Fellowship #1136550.