A Cerebellar Window for Intravital Imaging of Normal and Disease States in Mice

Overview

Journal Nat Protoc

Specialties Biology
Pathology
Science

Date 2017 Oct 6

PMID 28981123

Citations 19

Authors

Vasileios Askoxylakis

Mark Badeaux

Sylvie Roberge

Ana Batista

Ned Kirkpatrick

Matija Snuderl

Zohreh Amoozgar

Giorgio Seano

Gino B Ferraro

Sampurna Chatterjee

Lei Xu

Dai Fukumura

Dan G Duda

Rakesh K Jain

Affiliations

Soon will be listed here.

Abstract

The cerebellum is a prominent part of the vertebrate hindbrain that is critically involved in the regulation of important body functions such as movement coordination, maintenance of balance and posture, and motor control. Here, we describe a cerebellar window that provides access to the mouse cerebellum for intravital imaging, thereby allowing for a detailed characterization of the dynamic processes in this region of the brain. First, the skull overlying the cerebellum is removed, and then the window is applied to the region of interest. Windows may be exchanged depending on the desired imaging modality. This technique has a variety of applications. In the setting of medulloblastoma, spontaneous or orthotopically implanted lesions can be imaged, and tumor morphology and size can be monitored using ultrasonography. Multiphoton laser-scanning microscopy (MPLSM) or optical-frequency-domain imaging (OFDI) can be applied for in vivo visualization and analysis of cellular and vascular structures in a variety of disease states, including malignancies and ataxia telangiectasia. This protocol describes a novel and rapid method for cerebellar window construction that can be set up in under an hour.

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References

Egeblad M, Ewald A, Askautrud H, Truitt M, Welm B, Bainbridge E . Visualizing stromal cell dynamics in different tumor microenvironments by spinning disk confocal microscopy. Dis Model Mech. 2008; 1(2-3):155-67. PMC: 2562195. DOI: 10.1242/dmm.000596. View

Kedrin D, Gligorijevic B, Wyckoff J, Verkhusha V, Condeelis J, Segall J . Intravital imaging of metastatic behavior through a mammary imaging window. Nat Methods. 2008; 5(12):1019-21. PMC: 2820719. DOI: 10.1038/nmeth.1269. View

Walk E, McLaughlin S, Coad J, Weed S . Use of high frequency ultrasound to monitor cervical lymph node alterations in mice. PLoS One. 2014; 9(6):e100185. PMC: 4067293. DOI: 10.1371/journal.pone.0100185. View

Niermann K, Fleischer A, Huamani J, Yankeelov T, Kim D, Wilson W . Measuring tumor perfusion in control and treated murine tumors: correlation of microbubble contrast-enhanced sonography to dynamic contrast-enhanced magnetic resonance imaging and fluorodeoxyglucose positron emission tomography. J Ultrasound Med. 2007; 26(6):749-56. DOI: 10.7863/jum.2007.26.6.749. View

Vakoc B, Lanning R, Tyrrell J, Padera T, Bartlett L, Stylianopoulos T . Three-dimensional microscopy of the tumor microenvironment in vivo using optical frequency domain imaging. Nat Med. 2009; 15(10):1219-23. PMC: 2759417. DOI: 10.1038/nm.1971. View

Holtmaat A, Bonhoeffer T, Chow D, Chuckowree J, de Paola V, Hofer S . Long-term, high-resolution imaging in the mouse neocortex through a chronic cranial window. Nat Protoc. 2009; 4(8):1128-44. PMC: 3072839. DOI: 10.1038/nprot.2009.89. View

Kamoun W, Chae S, Lacorre D, Tyrrell J, Mitre M, Gillissen M . Simultaneous measurement of RBC velocity, flux, hematocrit and shear rate in vascular networks. Nat Methods. 2010; 7(8):655-60. PMC: 2921873. DOI: 10.1038/nmeth.1475. View

Helmlinger G, Yuan F, Dellian M, Jain R . Interstitial pH and pO2 gradients in solid tumors in vivo: high-resolution measurements reveal a lack of correlation. Nat Med. 1997; 3(2):177-82. DOI: 10.1038/nm0297-177. View

Wei S, Fattet L, Tsai J, Guo Y, Pai V, Majeski H . Matrix stiffness drives epithelial-mesenchymal transition and tumour metastasis through a TWIST1-G3BP2 mechanotransduction pathway. Nat Cell Biol. 2015; 17(5):678-88. PMC: 4452027. DOI: 10.1038/ncb3157. View

10.

Vuong B, Skowron P, Kiehl T, Kyan M, Garzia L, Sun C . Measuring the optical characteristics of medulloblastoma with optical coherence tomography. Biomed Opt Express. 2015; 6(4):1487-501. PMC: 4399685. DOI: 10.1364/BOE.6.001487. View

11.

Ager E, Kozin S, Kirkpatrick N, Seano G, Kodack D, Askoxylakis V . Blockade of MMP14 activity in murine breast carcinomas: implications for macrophages, vessels, and radiotherapy. J Natl Cancer Inst. 2015; 107(4). PMC: 4402365. DOI: 10.1093/jnci/djv017. View

12.

Chow D, Groszer M, Pribadi M, Machniki M, Carmichael S, Liu X . Laminar and compartmental regulation of dendritic growth in mature cortex. Nat Neurosci. 2009; 12(2):116-8. PMC: 2842592. DOI: 10.1038/nn.2255. View

13.

Goldey G, Roumis D, Glickfeld L, Kerlin A, Reid R, Bonin V . Removable cranial windows for long-term imaging in awake mice. Nat Protoc. 2014; 9(11):2515-2538. PMC: 4442707. DOI: 10.1038/nprot.2014.165. View

14.

Wyckoff J, Wang Y, Lin E, Li J, Goswami S, Stanley E . Direct visualization of macrophage-assisted tumor cell intravasation in mammary tumors. Cancer Res. 2007; 67(6):2649-56. DOI: 10.1158/0008-5472.CAN-06-1823. View

15.

Bovenberg M, Hannah Degeling M, Tannous B . Enhanced Gaussia luciferase blood assay for monitoring of in vivo biological processes. Anal Chem. 2011; 84(2):1189-92. PMC: 3264838. DOI: 10.1021/ac202833r. View

16.

Goel S, Gupta N, Walcott B, Snuderl M, Kesler C, Kirkpatrick N . Effects of vascular-endothelial protein tyrosine phosphatase inhibition on breast cancer vasculature and metastatic progression. J Natl Cancer Inst. 2013; 105(16):1188-201. PMC: 3748004. DOI: 10.1093/jnci/djt164. View

17.

Samano A, Ohshima-Hosoyama S, Whitney T, Prajapati S, Kilcoyne A, Taniguchi E . Functional evaluation of therapeutic response for a mouse model of medulloblastoma. Transgenic Res. 2010; 19(5):829-40. PMC: 4770905. DOI: 10.1007/s11248-010-9361-1. View

18.

Chauhan V, Martin J, Liu H, Lacorre D, Jain S, Kozin S . Angiotensin inhibition enhances drug delivery and potentiates chemotherapy by decompressing tumour blood vessels. Nat Commun. 2013; 4:2516. PMC: 3806395. DOI: 10.1038/ncomms3516. View

19.

Nakasone E, Askautrud H, Kees T, Park J, Plaks V, Ewald A . Imaging tumor-stroma interactions during chemotherapy reveals contributions of the microenvironment to resistance. Cancer Cell. 2012; 21(4):488-503. PMC: 3332002. DOI: 10.1016/j.ccr.2012.02.017. View

20.

Snuderl M, Batista A, Kirkpatrick N, Ruiz de Almodovar C, Riedemann L, Walsh E . Targeting placental growth factor/neuropilin 1 pathway inhibits growth and spread of medulloblastoma. Cell. 2013; 152(5):1065-76. PMC: 3587980. DOI: 10.1016/j.cell.2013.01.036. View