Antibody stabilization for thermally accelerated deep immunostaining.

Antibodies have diverse applications due to their high reaction specificities but are sensitive to denaturation when a higher working temperature is required. We have developed a simple, highly scalable and generalizable chemical approach for stabilizing off-the-shelf antibodies against thermal and chemical denaturation. We demonstrate that the stabilized antibodies (termed SPEARs) can withstand up to 4 weeks of continuous heating at 55 °C and harsh denaturants, and apply our method to 33 tested antibodies. SPEARs enable flexible applications of thermocycling and denaturants to dynamically modulate their binding kinetics, reaction equilibrium, macromolecular diffusivity and aggregation propensity. In particular, we show that SPEARs permit the use of a thermally facilitated three-dimensional immunolabeling strategy (termed ThICK staining), achieving whole mouse brain immunolabeling within 72 h, as well as nearly fourfold deeper penetration with threefold less antibodies in human brain tissue. With faster deep-tissue immunolabeling and broad compatibility with tissue processing and clearing methods without the need for any specialized equipment, we anticipate the wide applicability of ThICK staining with SPEARs for deep immunostaining.

High white matter hyperintensity burden in strategic white matter tracts relates to worse global cognitive performance in community-dwelling individuals.

BACKGROUND: White matter hyperintensities (WMH) are associated with cognitive impairment. The impact of WMH on cognitive domains (e.g. processing speed, executive functioning) depends on location. We determined whether the relevance of WMH location also applies to global cognitive functioning by testing if WMH in strategic white matter tracts are associated with global cognitive functioning independent of total WMH burden. METHODS: We included 830 community-dwelling individuals. WMH volume within two a priori specified strategic white matter tracts (forceps minor and anterior thalamic radiation) were entered in a linear regression model with the Montreal Cognitive Assessment (MoCA) and Mini-Mental State Examination (MMSE) as outcome variables and corrected for total WMH volume and other MRI markers for vascular injury and neurodegenerations (i.e. brain parenchymal fraction, and the presence of lacunes and microbleeds). RESULTS: WMH in the forceps minor and left anterior thalamic radiation inversely correlated with MoCA, and WMH in the forceps minor inversely correlated with MMSE, independent of total WMH volume and other MRI markers. CONCLUSION: The impact of WMH on global cognitive functioning depends on location. Whether this reflects accumulated impairment in isolated cognitive domains or disruption of a network that is crucially involved in global cognitive performance remains to be determined.

Peak width of skeletonized mean diffusivity and its association with age-related cognitive alterations and vascular risk factors.

INTRODUCTION: Only two studies investigated the associations between peak width of skeletonized mean diffusivity (PSMD) and age-related cognitive alterations, whereas none of the studies investigated the association with vascular risk factors. METHODS: We evaluated 801 stroke- and dementia-free elderlies with baseline and 3-year follow-up assessments. Regression analyses were used to assess the association between age-related cognitive functions and PSMD. Simple mediation models were used to study the mediation effect of PSMD between vascular risk factors and age-related cognitive outcomes. RESULTS: PSMD was negatively associated with processing speed at baseline and negatively associated with processing and memory scores at 3-year follow-up. The association between vascular risk factors and age-related cognition was mediated by PSMD, as well as other diffusion tensor imaging markers. DISCUSSION: PSMD is preferred over other diffusion tensor imaging markers as it is sensitive to age-related cognitive alterations and calculation is fully automated. PSMD is proposed as a research tool to monitor age-related cognitive alterations.