Fluid Aspiration from Mouse Glomeruli: A Method to Aspirate Glomerular Fluid from Mouse Using Two-Photon Mediated Micropuncture

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.

1. Weigh the mouse.
2. Induce anesthesia using 4% isoflurane and maintain with 1.5–2.5% isoflurane in air/oxygen mixture. Confirm that t....

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 sh.......

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
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