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

タグ

Play Video

記事を引用
Fluid Aspiration from Mouse Glomeruli: A Method to Aspirate Glomerular Fluid from Mouse Using Two-Photon Mediated Micropuncture. J. Vis. Exp. (Pending Publication), e20577, doi: (2023).

View Video