Fluid Aspiration from Mouse Glomeruli: A Method to Aspirate Glomerular Fluid from Mouse Using Two-Photon Mediated Micropuncture
Published: April 30, 2023
Abstract
Source: Matsushita, K. et al. Micropuncture of Bowman's Space in Mice Facilitated by 2 Photon Microscopy. J. Vis. Exp. (2018)
This video showcases fluid aspiration from mouse glomeruli using the two-photon mediated micropuncture technique. The technique provides access to the living mouse’s glomeruli and thus helps gain insights into renal physiology.
Protocol
All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board. 1. Lateral Pipette Access to The Kidney Below a Fluid Imaging Column Via a Novel Surgical Procedure NOTE: The assembly of the imaging support system and surgical prep is shown in Figure 1. The procedure described is performed on C57BL/6 mice weighing 20–25 g. <…
Representative Results
Figure 1: Partial extrusion of the kidney with custom support and immobilization for lateral access. On the left, the parts of the imaging column and kidney support are shown, with the complete assembly in center. On the right, the kidney preparation is shown before (above) and after (below) application of the support. <img alt="Figure…
開示
The authors have nothing to disclose.
Materials
| Upright 2 photon microscope | Zeiss | LSM 7MP | |
| 3 axis microscope stage controller | Sutter | MP-285 | |
| 3 axis headstage controller | Sutter | MP-225 | |
| Pipette holder | Molecular Devices | 1-HL-U | |
| Headstage | Molecular Devices | CV203BU | |
| FITC-dextran 2000 kDa MW | Sigma-Aldrich | 52471-1G | |
| Borosilicate glass capillary tubes | Sutter | B150-110-7.5 | |
| Micropipette puller | Sutter | P-97 | |
| Quantum dots, 605 nm | Thermofisher | Q21701MP | |
| Polysiloxane | Sugru | www.sugru.com, "original formula". Any color. | |
| PE-50 tubing | Instech Labs | BTPE-50 | |
| Microinjector | WPI | UMP-3 | |
| Microinjector controller | WPI | Micro4 | |
| Perfluorodecalin | Sigma-Aldrich | 306-94-5 | |
| Agarose | Sigma-Aldrich | 9012-36-6 | |
| Coverslip, 10 mm | Harvard Apparatus | 64-0718 | |
| Headplate | Custom | Common in neuroscience labs, many suppliers | |
| Head fixation device | Custom | Common in neuroscience labs, many suppliers | |
| 30G needle | Becton-Dickinson | 125393 | For retroorbital injection |
| Tuberculin syringe | Becton-Dickinson | 309626 | For retroorbital injection |
