Abstract
Ischemic stroke is a leading cause of death and permanent disability worldwide. Middle cerebral artery occlusion (MCAO) of variable duration times could be anticipated to result in varying degrees of injury that evolve spatially over time. Therefore, investigations following strokes require information concerning the spatiotemporal dimensions of the ischemic core as well as of perilesional areas. In the present study, multiphoton microscopy (MPM) based on two-photon excited fluorescence (TPEF) and second harmonic generation (SHG) was applied to image such pathophysiological events. The ischemic time-points for evaluation were set at 6, 24, 48, and 72 hours after MCAO. Our results demonstrated that MPM has the ability to not only identify the normal and ischemic brain regions, but also reveal morphological changes of the cortex and striatum at various times following permanent MCAO. These findings corresponded well with the hematoxylin and eosin (H&E) stained tissue images. With the technologic progression of miniaturized imaging devices, MPM can be developed into an effective diagnostic and monitoring tool for ischemic stroke.
Highlights
Stroke is a leading cause of death and permanent disability worldwide, in which ischemic injury results from vascular occlusion and a subsequent cerebral infarction [1, 2]
In examining all of the hematoxylin and eosin (H&E) stained sections, the core of the ischemic infarct was clearly detectable at all time-points investigated after permanent Middle cerebral artery occlusion (MCAO), and the infarct size depended on time of occlusion duration
The severity of neuronal injury and occlusion duration were directly related. These results demonstrated that two-photon excited fluorescence (TPEF) microscopy can be used to evaluate the structural changes of injured neurons at various ischemic time-points, which were essential to understanding the basic pathophysiology of cerebrovascular ischemia, as well as to evaluating the effect of treatment regimens. 3.4 Structural alterations in the striatum Ischemic injury to the striatum was examined in addition to the cerebral cortex
Summary
Stroke is a leading cause of death and permanent disability worldwide, in which ischemic injury results from vascular occlusion and a subsequent cerebral infarction [1, 2]. Necrotic death mainly affects cells in the ischemic core and delayed death impacts susceptible neurons in neighboring regions. As cell death in injury core-adjacent regions occurs over an extended period of time, these neurons may be targeted for rescue by pharmacological agents and other therapies. Spatial and temporal identification of the boundaries of the infarct core as well as the discrimination of differentially affected perilesional areas is essential to investigate correlations between the extent of neuroprotection or the variability of post-ischemic gene regulation with a defined degree of injury [9,10,11]
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