Method for establishing analysis module for assessing neurological functions

Abstract

The invention provides a method for establishing an analysis module for assessing neurological functions. The method comprises the following steps: acquiring culture cells which include a plurality of fluorescent labels by virtue of a micro-fluorescent imaging system so as to conduct image analysis, wherein the culture cells include nerve cells and non-nerve cells; in accordance with area size of cell nucleuses and fluorescence intensity, selecting the nerve cells showing axon and dendrite fluorescent labels, eliminating non-nerve cells, and calculating the area sizes of the nerve cell bodies as well as axon and dendrite lengths and the number of branches, so that axon and dendrite growth situations of the nerve cells are determined; and in accordance with axon and dendrite fluorescent labeling ranges, calculating the number of synaptic puncta fluorescent labels on the nerve cells, so that the growth situation of the nerve cells is determined, and the neurological functions are assessed. The method disclosed by the invention can further serve as a drug screening platform, so as to rapidly detect whether drugs have a protective or neurotoxic effect on the nerve cells or not.

Description

technical field [0001] The present invention relates to a method for establishing an analysis module for evaluating nerve function, in particular to a set of rapid and representative methods for analyzing nerve growth function, including using important markers sufficient to represent nerve growth, and matching with optimized Image analysis process and program instructions; accordingly, this method can not only quickly evaluate the early nerve growth and neural network formation, but also serve as a platform for screening drugs to quickly detect the effects of drugs on nerve function. Background technique [0002] With the advancement of cancer treatment, the survival rate of patients has been gradually improved. Due to the prolongation of the patient's life expectancy, the side effects of chemotherapy drugs have been paid more attention, especially among the side effects of chemotherapy drugs, neuropathy has the greatest impact on the quality of life. Therefore, before rece...

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Problems solved by technology

However, there are still many problems in the current method of using molecular markers combined with high-throughput analysis to detect neural function; for example, Huang Y et al. revealed a high content neuron-based screening in vitro to explore the A morphological computational framework that uses neuronal type III β-tubulin (TUJ1) as a molecular marker to study neuronal mechanisms (J Neurosci Methods.2010Jul 15; 190(2):299-309); Harrill JA et al Revealed a high-content image analysis to detect the effects of chemicals on the growth of primary rat cortical neuron axons and dendrites, using tubulin (β III After deducting the microtubule-associated protein 2 (MAP2) staining area (dendrites) from the tubulin) staining area (all), the remaining area represents the axon, and the growth of nerve cells is analyzed (Neurotoxicology.2013Jan; 34:61-73 ); Nieland TJ et al. used MAP2, postsynaptic density protein 95-synapsin 1 (Psd95-Syn1), and postsynaptic protein-synapsin 1 (Gphn-Syn1) to identify synaptogenesis status (PLoS One.2014Mar 14; 9(3):e91744); the above-mentioned method can not comprehensively judge whether the nerve function is good or bad only by means of outgrowth or synaptogenesis; The axons, dendritic growth and synapse formation of nerve cells require the use of multiple molecular markers, so the process is quite tedious and time-consuming

[0004] Furthermore, what kind of nerve cells are used for drug testing will also affect the analysis procedure and time; for example, although commercial nerve cell lines are better cultivated, in order to stimulate nerve cell differentiation, additional drug stimulation must be added, which may affect the time to be tested. Measuring the effect of drugs

In contrast, primary culture cells can naturally differentiate, so the use of primary culture cells can solve the problem of adding additional drugs, but compared with using cell lines, primary cell culture is not easy, and how to obtain the best differentiation time point for drug testing, still need to be further explored

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