Anmelden

Max Planck Institute of Experimental Medicine

2 ARTICLES PUBLISHED IN JoVE

Neuroscience

Genetic Manipulation of Cerebellar Granule Neurons In Vitro and In Vivo to Study Neuronal Morphology and Migration

Anna Holubowska^1,2, Chaitali Mukherjee^1,2, Mayur Vadhvani^1,2, Judith Stegmüller^1,2

¹Cellular and Molelcular Neurobiology, Max Planck Institute of Experimental Medicine, ²Center for Nanoscale Microscopy and Molecular Physiology of the Brain (CNMPB)

Neuronal morphogenesis and migration are crucial events underlying proper brain development. Here, we describe methods to genetically manipulate cultured cerebellar granule neurons and the developing cerebellum for the assessment of morphology and migratory characteristics of neurons.

Biology

Biological Sample Preparation by High-pressure Freezing, Microwave-assisted Contrast Enhancement, and Minimal Resin Embedding for Volume Imaging

Anna M. Steyer^1,2, Torben Ruhwedel¹, Wiebke Möbius^1,2

¹Electron Microscopy Core Unit, Max Planck Institute of Experimental Medicine, ²Center Nanoscale Microscopy and Molecular Physiology of the Brain (CNMPB)

Here we present a protocol for combining two sample processing techniques, high-pressure freezing and microwave-assisted sample processing, followed by minimal resin embedding for acquiring data with a focused ion beam scanning electron microscope (FIB-SEM). This is demonstrated using a mouse tibial nerve sample and Caenorhabditis elegans.