Overview

Maintaining and controlling cell shape, cell movement, cytokinesis and the organization of organelles is one of the most basic functions of the cell. Because change in these features is often the consequence of cellular differentiation, cellular toxicity, pathology or other critical cellular event or signaling, their measurement against potential therapeutic targets can be critical. The cytoskeleton is typically composed of microfilaments, microtubules, and intermediate filaments in the cell and plays a key role in the morphological change of cells. The cytoskeleton also facilitates proper function of other cellular proteins through direct binding, transporting, repositioning and sequestering these proteins. For certain cell types such as neurons, this morphological change of the cell is indispensable to gain the proper function in the tissue.

Fluorescence microscopy and imaging, the basis of High Content Screening (HCS), High Content Analsis (HCA) and image cytometry provide both intensity as well as spatial information of the fluorescently-labeled constituents that are imaged and quantitatively analyzed. Spatial information, such as cell shape and morphology, sizes and arrangements of intracellular constituents, and the arrangement and location of cells relative to each other, can all be obtained from fluorescence microscopy and HCS. Other cell-based fluorescence assay methods such as flow cytometry, FLIPR, and homogeneous fluorescence signals in plate readers cannot provide this type of high-content data.

Neurite Morphology

The synapse consists of two main structures: the presynaptic compartment that releases neurotransmitters and the postsynaptic compartment where these neurotransmission signals are received. Synaptic damage has often been recognized as the first sign of neurodegeneration in many different pathological conditions, including traumatic nerve injury, ischemic stroke, and neurodegenerative disorders such as Motor Neuron Disease, Alzheimer’s, Parkinson’s and Huntington’s diseases as many synaptic proteins play an important role in the progression of neurodegenerative diseases. Since modulation of neurite and synaptic structures in neurons is closely related to the pathological process of neurological diseases or in neurodevelopment, synapse number in neurons can be a sensitive indicator for the change of neuronal function.

Cell Cycle and Cytoskeleton
Cytoskeletal rearrangement and cell cycle progression can serve as sensitive indicators of cell growth and cell stress. Thermo Scientific have developed several multiplexed, quantitative, cell based HCS assays to monitor these key cellular functions. The Thermo Scientific Cellomics Cytoskeletal Rearrangement HCS reagents simultaneously detect changes in cytoskeletal structures (F-actin and microtubule fibers), nuclear DNA content, and nuclear and cellular morphologies using bright fluorescent probes. The Thermo Scientific Cellomics Cell Cycle I HCS Reagents characterize cell cycle progression by simultaneously quantifying DNA content, BrdU incorporation and Histone H3 phosphorylation. By assessing the cell cycle using BrdU and phospho-Histone H3 antibodies while simultaneously monitoring cytoskeletal rearrangements and cell morphological changes, high-conent can establish a multiparameter profile of different drugs affecting cell cycle, cell growth, cell differentiation and cell death.