Name: pancreatic intravital imaging window installation in mouse model: an integrated pancreatic imaging window for time-lapse in vivo imaging of murine pancreas microstructure
Uploaded: 2023-04-30T14:51:22.000+00:00
Duration: 1 min 45 s
Description: Source: Park, I. et. al. Stabilized Longitudinal In Vivo Cellular-Level Visualization of the Pancreas in a Murine Model with a Pancreatic Intravital ...

pancreatic intravital imaging window installation in mouse model: an integrated pancreatic imaging window for time-lapse in vivo imaging of murine pancreas microstructure

Pancreatic Intravital Imaging Window Installation in Mouse Model: An Integrated Pancreatic Imaging Window for Time-lapse In Vivo Imaging of Murine Pancreas Microstructure

Begin by preparing the surgical platform, and disinfecting the surfaces with 70% ethanol. After anesthetization, use a rectal probe with a homeothermic controlled heating pad to monitor the body temperature.
Remove hair from the left flank of the mouse and apply alcohol and iodine alternatively, rotating from the incision site to the outer surface, never going back to the incision site and ending with iodine. Then make a 1.5-centimeter incision on the left flank of the mouse dissecting the skin and the muscle.
Use a black or nylon 4-0 suture to perform a purse-string suture in the incision margin. Pool the spleen carefully with ring forceps, and identify the pancreas. Placing the window at the flank of the mouse, pass the spleen and the pancreas through the open space of the window.
Then, gently place the pancreas on the plate of the imaging window while placing the spleen on the open space of the window. Use a 31 gauge catheter needle to apply drops of N-butyl cyanoacrylate glue to the margin of the imaging window, ensuring a minimal amount of glue is applied.
Gently apply a 12-millimeter round cover glass to the margin of the imaging window. Then, hold the suture loop to fit into the lateral groove of the window and tie it three times. Finally, to prevent the interruption of these tight stitches when the mice are awake, cut the maximal proximal site of the tie.

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All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.
1. Surgery
<ol>
	<li>Prepare a sterile surgical platform and sterilize the surfaces with 70% ethanol.| 
	NOTE: For longitudinal imaging sessions, aseptic technique is essential.</li>
	<li>Anesthetize mice with a mixture of tiletamine/zolazepam (30 mg/kg) and xylazine (10 mg/kg). 
	NOTE: Use of tiletamine/zolazepam is recommended instead of ketamine because of its adverse effect of hyperglycemia. Optimal anesthesia should be selected for the purpose of the experiment.</li>
	<li>Monitor the body temperature using a rectal probe with a homeothermic controlled heating pad.</li>
	<li>Shave the left flank of the mouse and apply three rounds of alternating alcohol and iodine-based scrub. 
	NOTE: If depilatory cream is used,&#160;do not leave longer than 1 minute to avoid chemical burn.</li>
	<li>Make a 1.5 cm incision on the left flank of the mouse and dissect the skin and muscle.</li>
	<li>Perform a purse-string suture with a black or nylon 4-0 suture in the incision margin.</li>
	<li>Use a micro retractor on the incision and gently expose the spleen.</li>
	<li>Carefully pool the spleen with ring forceps and identify the pancreas.</li>
	<li>Place the window at the flank of the mouse and pass the spleen and pancreas through the open space of the window.&#160;Manipulation of the pancreas has to be very gentle as it can cause bleeding or pancreatitis. 
	NOTE: Custom design the pancreatic intravital imaging window to seclude the pancreas from the bowel in the abdominal cavity (Figure 1A, B).</li>
	<li>Gently place the pancreas on the plate of the imaging window; the spleen will be placed on the open space of the window.</li>
	<li>Apply drops of N-butyl cyanoacrylate glue on the margin of the imaging window.
	<ol>
		<li>To minimize the amount of the applied glue, use a 31 G catheter needle for the application. If the amount of the drop is large, then the tissue will unintentionally adhere to the window or cover glass.</li>
	</ol>
	</li>
	<li>Gently apply a 12 mm round cover glass to the margin of the imaging window.</li>
	<li>Pull the suture loop to fit into the lateral groove of the window and tie it three times.</li>
	<li>Cut the maximal proximal site of the knot&#160;to prevent the interruption of tight stitches when these mice are awake.</li>
	<li>Let mice recover from anesthesia (2-3 hours) and inject ketoprofen (5 mg/kg, subcutaneous in the loose skin at the base of neck or hip of the animal) for pain relief. Reassess for signs of pain every 12 hours and&#160;ketoprofen should be considered every 24 hours for 1-3 days. 
	NOTE: Analgesia influences insulin secretion in response to glucose. The choice and timing of analgesia must be individualized for the experimental purpose.</li>
	<li>Accommodate the mouse in the cage with sufficient bedding to avoid the contact of the window to the cage. Avoid housing the house with the mice without the window surgery.</li>
</ol>

<table><tbody><tr><td>Cover glasses circular</td><td>Marienfeld</td><td>111520</td><td>Cover glass for pancreatic imaging window</td></tr><tr><td>IRIS Scissor</td><td>JEUNGDO BIO &amp;amp; PLANT CO, LTD</td><td>S-1107-10</td><td>This product can be replaced with the product from other company</td></tr><tr><td>Loctite 401</td><td>Henkel</td><td>401</td><td>N-butyl cyanoacrylate glue</td></tr><tr><td>Micro Needle holder</td><td>JEUNGDO BIO &amp;amp; PLANT CO, LTD</td><td>H-1126-10</td><td>This product can be replaced with the product from other company</td></tr><tr><td>Micro rectractor</td><td>JEUNGDO BIO &amp;amp; PLANT CO, LTD</td><td>17004-03</td><td>This product can be replaced with the product from other company</td></tr><tr><td>Microforceps</td><td>JEUNGDO BIO &amp;amp; PLANT CO, LTD</td><td>F-1034</td><td>This product can be replaced with the product from other company</td></tr><tr><td>Nylon 4-0</td><td>AILEE</td><td>NB434</td><td>Non-Absorbable Suture</td></tr><tr><td>Omnican N 100 30G</td><td>B BRAUN</td><td>FT9172220S</td><td>For Vascular Catheter, Use only Needle part</td></tr><tr><td>Pancreatic imaging window</td><td>Geumto Engineering</td><td>Custom order</td><td>Pancreatic imaging window - custom order</td></tr><tr><td>Physiosuite</td><td>Kent Scientific</td><td>PS-02</td><td>Homeothermic temperature controller</td></tr><tr><td>Ring Forceps</td><td>JEUNGDO BIO &amp;amp; PLANT CO, LTD</td><td>F-1090-3</td><td>This product can be replaced with the product from other company</td></tr><tr><td>Rompun</td><td>Bayer</td><td>Rompun</td><td>Anesthetic agent</td></tr><tr><td>Zoletil</td><td>Virbac</td><td>Zoletil 100</td><td>Anesthetic agent</td></tr><tr><td>BALB/C Nude&amp;nbsp;</td><td>OrientBio&amp;nbsp;</td><td>BALB/C Nude</td><td>BALB/C Nude</td></tr></tbody></table>

Source: Park, I. et. al. <a href="https://www.jove.com/v/62538">Stabilized Longitudinal In Vivo Cellular-Level Visualization of the Pancreas in a Murine Model with a Pancreatic Intravital Imaging Window.</a> J. Vis. Exp. (2021)
This video demonstrates the procedure to implant a pancreatic imaging window to separate the pancreas from the intestines for longitudinal time-lapse high-resolution imaging of the pancreas microstructure.

<img alt="Figure 1" src="/files/ftp_upload/20796/20796_Figure1.jpg" /> 
Figure 1. Design and photograph of the pancreatic intravital imaging window. (A) A 3D and cross-sectional view of the pancreatic intravital imaging window. (B) Anterior and posterior photograph of the pancreatic imaging window.

Source: Park, I. et. al. Stabilized Longitudinal In Vivo Cellular-Level Visualization of the Pancreas in a Murine Model with a Pancreatic Intravital ...

High-resolution intravital microscopy, via an implanted pancreatic imaging window, facilitates long-term, wide-area cellular-level visualization of the pancreatic islets in a live animal.&#160;
For pancreatic imaging window implantation, first, position an anesthetized mouse in a left lateral position. Next, fix a probe to monitor the mouse&#8217;s temperature and regulate its temperature using a heating pad. Now, shave the left flank and sterilize the area.
Make a surgical incision and dissect the abdominal muscles. Insert appropriate sutures in the incision margin to affix the imaging window frame in position following implantation. Further, expose and exteriorize the spleen, and identify the connected pancreas.
Next, overlay the pancreas connected with the spleen on the plate of the customized pancreatic imaging window, with the spleen positioned in the open space of the window. This positioning of the pancreas effectively isolates it from the neighboring bowel movements, reducing the motion-induced artifacts during imaging. Carefully apply cyanoacrylate glue to the margin of the imaging window, avoiding the organs.
Mount a cover glass over the margin of the imaging window. Tighten the sutures around the imaging window to secure the window in place. The mouse with the implanted pancreatic imaging window is ready for intravital pancreatic imaging.

1.2K ビュー. マウスモデルにおける膵臓生体内イメージングウィンドウの設置:マウス膵臓微細構造のタイムラプスIn vivoイメージングのための統合膵臓イメージングウィンドウ

Pancreatic Intravital Imaging Window Installation in Mouse Model: An Integrated Pancreatic Imaging Window for Time-lapse In Vivo Imaging of Murine Pancreas Microstructure

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Pancreatic Intravital Imaging Window Installation in Mouse Model: An Integrated Pancreatic Imaging Window for Time-lapse In Vivo Imaging of Murine Pancreas Microstructure