Method for assessment of neural function by establishing analysis module

A method for identifying and assessing function of neurons in a co-culture, comprising:

• providing a co-culture containing neurons and non-neuronal cells;
• obtaining a first fluorescence signal identifying DNA of the neurons and the non-neuronal cells in the co-culture;
• a second fluorescence signal identifying neurites of the neurons in the co-culture;
• and a third fluorescence signal identifying synaptic puncta of the neurons in the co-culture;
• identifying the neurons in the co-culture by obtaining identity information including steps (a) to (c):
• (a) DNA of the neurons and the non-neuronal cells identified by a fluorescence intensity of the first fluorescence signal;
• (b) dimensions of an area containing the DNA of the neurons and the non-neuronal cells identified by an area of the first fluorescence signal;
• and
• (c) the DNA of the neurons and the non-neuronal cells, the dimensions of an area containing the DNA, and the neurites of the neurons identified by a merging comparison of steps (a) and (b);
• wherein the identified neurons are selected and the non-neuronal cells are excluded as noise by performance of steps (a) to (c);
• after identifying and selecting the neurons and excluding the non-neuronal cells, function of the identified neurons is assessed by:
• conducting a neuronal neurite outgrowth assessment by executing an outgrowth module on the identified and selected neurons using the second fluorescence signal and the identity information, wherein the outgrowth module measures outgrowth of an area of a neuron body, a length of a process, a number of process, branches of a neuron or a combination thereof;
• and
• conducting a synaptic puncta assessment by executing a cell scoring and transfluor module on the identified and selected neurons using the second fluorescence signal, the third fluorescence signal and the identity information, wherein the cell scoring and transfluor module measures outgrowth of synaptic puncta by a location of the synaptic puncta along with a length of a process,
• wherein performance of the outgrowth module and the cell scoring and transfluor module together provide assessment of the function of the identified neurons.