Analyzing neurite outgrowth, comprises depositing neuron and neuron-like cell on substrate having grid pattern with first- and second region on which neuron and neuron-like cell accumulate, where first region is surrounded by second region

摘要

The method comprises depositing neurons and neuron-like cells on a substrate having a grid pattern, which has first- (1) and second regions (2), on which the neurons and neuron-like cells accumulate, where the first regions are surrounded by second regions, and subsequently exposing the neurons and neuron-like cells to test substances during which and/or after the neurite outgrowth is analyzed from the neurons and neuron-like cells. The substrate has first region with standardized parameter and position. The first regions are equidistantly spaced from each other. The method comprises depositing neurons and neuron-like cells on a substrate having a grid pattern, which has first- (1) and second regions (2), on which the neurons and neuron-like cells accumulate, where the first regions are surrounded by second regions, and subsequently exposing the neurons and neuron-like cells to test substances during which and/or after the neurite outgrowth is analyzed from the neurons and neuron-like cells. The substrate has first region with standardized parameter and position. The first regions are equidistantly spaced from each other, and are arranged in a hexagonal pattern and/or a linear pattern. The first regions are circular, have a diameter of 20-200 mu m, and are separated from each other in a distance of 50-1000 mu m. The substrate is used, in which the adjacent first regions are bound with one another through paths, where the width of the paths is 100 nm to 5 mu m. The first regions are formed hydrophilic and the second regions are formed hydrophobic. The neurons and neuron-like cells are deposited on the substrate without fixation and marking and are subsequently analyzed, are fixed at the substrate using standard methods, and are completely or partially marked. The neurite outgrowth is measured by measurements such as number of neurites, number of viable neurites, number of cells possessing neurites, number of neurites per cell, neurite length, neurite growth rate, neurite branching and number of neurite per viable cell. The measurements of the neurite growth are carried out continuously, periodically or at the end of the exposition time period of the test substance. The substrate is disposed in a chamber for standard tissue culture or molecular analysis including 6-, 12-, 24-, 96-, 384- and 1536-well plates. The chamber for standard tissue culture or molecular analysis including 6-, 12-, 24-, 96-, 384- and 1536-well plates is provided with first and second regions corresponding to the substrate for depositing or undepositing the neurons and neuron-like cells. The liquids and test substances are supplied or removed using pipettes, automated systems for handling liquids or microfluidic systems.