Ex Vivo Imaging of Postnatal Cerebellar Granule Cell Migration Using Confocal Macroscopy

Samenvatting

Source: Bénard, M., et al. Ex Vivo Imaging of Postnatal Cerebellar Granule Cell Migration Using Confocal Macroscopy. J. Vis. Exp. (2015)

This video demonstrates a confocal macroscopic method to track the ex vivo migration of granule cells in cerebellar slices from postnatal rat brains.

Protocol

All procedures involving animal samples have been reviewed and approved by the appropriate animal ethical review committee.

1. Fluorescent Staining of Living Interneurons

1. Transfer cerebellar slices with a wide-bore glass truncated Pasteur pipette to a 6-well plate (max 3 slices/per well). Aspirate the Hanks’ balanced salt solution (HBSS) medium.

2. Incubate slices (3 max) in 5 ml of loading solution of the fluorescent dye (10 μM).

3. To protect from light, cover the microplate with aluminum foil. Put it on a gyro-moving table at 35 rpm for 10 min at RT to fa....

Resultaten

Figure 1: Ex vivo culture of P10 cerebellar slices. (A<.......

Materialen

Lijst van materialen gebruikt in dit artikel

Naam	Bedrijf	Catalogusnummer	Opmerkingen
Dulbecco’s modified eagle medium (DMEM) nutrient mixture F-12	Sigma-Aldrich	D8437
Cell Tracker Green CMFDA	Invitrogen	C2925
Polyester Transwell-Clear inserts	Corning	3452
6-well Cell culture cluster	Corning	3516
DMSO	Fisher Scientific	BP231-100
Gyro-rocker, SSL3	Stuart
CO₂ incubator, Hera Cell 150	Thermo Scientific
Confocal macroscope, TCS LSI	Leica Microsystems
CO₂-controller	PeCon
Hank's balanced salt solution 10x	Sigma-Aldrich	H1641
Adjustable-volume pipette (0.5-10μL)	Eppendorf	4910 000.018
Temperature controller	PeCon