A subscription to JoVE is required to view this content. Sign in or start your free trial.
In vivo high-resolution imaging of the pancreas was facilitated with the pancreatic intravital imaging window.
Direct in vivo cellular-resolution imaging of the pancreas in a live small animal model has been technically challenging. A recent intravital imaging study, with an abdominal imaging window, enabled visualization of the cellular dynamics in abdominal organs in vivo. However, due to the soft sheet-like architecture of the mouse pancreas that can be easily influenced by physiologic movement (e.g., peristalsis and respiration), it was difficult to perform stabilized longitudinal in vivo imaging over several weeks at the cellular level to identify, track, and quantify islets or cancer cells in the mouse pancreas. Herein, we describe a method for implanting a novel supporting base, an integrated pancreatic intravital imaging window, that can spatially separate the pancreas from the bowel for longitudinal time-lapse intravital imaging of the pancreas microstructure. Longitudinal in vivo imaging with the imaging window enables stable visualization, allowing for the tracking of islets over a period of 3 weeks and high-resolution three-dimensional imaging of the microstructure, as evidenced here in an orthotopic pancreatic cancer model. With our method, further intravital imaging studies can elucidate the pathophysiology of various diseases involving the pancreas at the cellular level.
The pancreas is an abdominal organ with an exocrine function in the digestive tract and an endocrine function of secreting hormones into the bloodstream. High-resolution cellular imaging of the pancreas could reveal the pathophysiology of various diseases involving the pancreas, including pancreatitis, pancreatic cancer, and diabetes mellitus1. Conventional diagnostic imaging tools such as computed tomography, magnetic resolution imaging, and ultrasonography are widely available in the clinical field1,2. However, these imaging modalities are restricted to visualizing only structural or ....
All procedures described in this paper were conducted in accordance with the 8th edition of the Guide for the Care and Use of Laboratory Animals (2011)26 and approved by the Institutional Animal Care and Use Committee at the Korea Advanced Institute of Science and Technology (KAIST) and Seoul National University Bundang Hospital (SNUBH).
1. Preparation of the window and other materials
Intravital microscopy combined with the supporting base integrated pancreatic intravital imaging window enables longitudinal cellular level imaging of the pancreas in a mouse. This protocol with the pancreatic intravital imaging window provides long-term tissue stability that enables the acquisition of high-resolution imaging to track individual islets for up to 3 weeks. As a result, mosaic imaging for an extended field of view, three-dimensional (3D) reconstruction of z-stack imaging, and longitudinal tracking of the sa.......
The protocol described here consists of intravital imaging of the pancreas using a novel supporting base integrated pancreatic intravital imaging window modified from an abdominal imaging window. Among the protocols described above, the first critical step is the implantation of the intravital pancreatic imaging window in the mouse. For the application of the glue in the window, it is important to apply the glue between the margin of the window and the cover glass, but not on the pancreatic tissue, as it may significantl.......
The authors have nothing to disclose.
This study was supported by grant No. 14-2020-002 from the SNUBH Research Fund and by the National Research Foundation of Korea (NRF) grant funded by the Korea government (MSIT) (NRF-2020R1F1A1058381, NRF-2020R1A2C3005694).
....Name | Company | Catalog Number | Comments |
Alexa Fluor 647 Succinimidyl Esters (NHS esters) | Invitrogen | A20006 | Fluorescent probe for conjugate with antibody |
BALB/C Nude | OrientBio | BALB/C Nude | BALB/C Nude |
BD Intramedic polyethylene tubing | BD Biosciences | 427401 | PE10 catheter for connection with needle |
C57BL/6N | OrientBio | C57BL/6N | C57BL/6N |
Cover glasses circular | Marienfeld | 0111520 | Cover glass for pancreatic imaging window |
FITC Dextran 2MDa | Merck (Former Sigma Aldrich) | FD200S | For vessel identification |
IMARIS 8.1 | Bitplane | IMARIS | Image processing |
Intravital Microscopy | IVIM tech | IVM-C | Intravital Microscopy |
IRIS Scissor | JEUNGDO BIO & PLANT CO, LTD | S-1107-10 | This product can be replaced with the product from other company |
Loctite 401 | Henkel | 401 | N-butyl cyanoacrylate glue |
Micro Needle holder | JEUNGDO BIO & PLANT CO, LTD | H-1126-10 | This product can be replaced with the product from other company |
Micro rectractor | JEUNGDO BIO & PLANT CO, LTD | 17004-03 | This product can be replaced with the product from other company |
Microforceps | JEUNGDO BIO & PLANT CO, LTD | F-1034 | This product can be replaced with the product from other company |
MIP-GFP | The Jackson Laboratory | 006864 | B6.Cg-Tg(Ins1-EGFP)1Hara/J |
Nylon 4-0 | AILEE | NB434 | Non-Absorbable Suture |
Omnican N 100 30G | B BRAUN | FT9172220S | For Vascular Catheter, Use only Needle part |
PANC-1 NucLightRed | Custom-made | Custom-made | Made in laboratory |
Pancreatic imaging window | Geumto Engineering | Custom order | Pancreatic imaging window - custom order |
Physiosuite | Kent Scientific | PS-02 | Homeothermic temperature controller |
Purified NA/LE Rat Anti-Mouse CD31 | BD Biosciences | 553708 | Antibody for in vivo vessel labeling |
Ring Forceps | JEUNGDO BIO & PLANT CO, LTD | F-1090-3 | This product can be replaced with the product from other company |
Rompun | Bayer | Rompun | Anesthetic agent |
TMR Dextran 65-85kDa | Merck (Former Sigma Aldrich) | T1162 | For vessel identification |
Window holder | Geumto Engineering | Custom order | Window holder - custom order |
Zoletil | Virbac | Zoletil 100 | Anesthetic agent |
Request permission to reuse the text or figures of this JoVE article
Request PermissionThis article has been published
Video Coming Soon
ABOUT JoVE
Copyright © 2025 MyJoVE Corporation. All rights reserved