Ultrathin CNS slices: Enhanced visualization of neuron structure for electrophysiology and imaging

Applications 1 | 2 | 3

Leica VT1000 S   Read the entire application report

Since the ability to resolve detailed cellular features is generally limited to 40-50 mm from the slice surface, we have developed an alternate strategy to visualize neurons using a semi-transparent ‘ultrathin’ slice preparation (20-50 mm) visualized on an inverted microscope equipped with Hoffman modulation optics. Isolated cerebellum, hippocampus or spinal cord from neonatal rats (P1- P14) were embedded in AGAR (2.5% w/v) then sectioned with a Leica VT1000E vibrating blade microtome. After incubation at 32°C, slices were fixed to the bottom of the recording chamber and maintained at room temperature. The enhanced transparency due to reduced slice thickness permitted superior optical resolution of dendritic and axonal processes and cells were visible throughout the slice thickness. Image: Live cell indicator reveals abundance of living cells throughout slice (P1 rat) (scale bar = 50 µm)