dic microscopy imaging  (Nikon)

Journal: Communications Chemistry

Article Title: Surface wetting is a key determinant of α-synuclein condensate maturation

doi: 10.1038/s42004-025-01764-z

Figure Lengend Snippet: a A diagram outlining the methodology used to monitor α-syn liquid condensate formation, sedimentation and maturation. Samples, loaded in a 96-well glass-bottom plate sealed with a clear film, are incubated at 37°C. Automated 3-step z-stack imaging in solution, alongside bottom-of-well imaging, is performed at selected time points over 20 h. Solution image analysis is then performed to quantify the number and area of all in focus objects within the image. b Representative DIC images acquired in solution during FL α-syn incubation. Scale bars represent 25 μm. c DIC microscopy-derived object area distribution over time. Each timepoint was compiled over three z-stack images. Mean areas are indicated by a solid black line. d Normalized total object count (left Y-axis) and normalized ThT intensity (right Y-axis) as a function of time. Three biological repeats are shown for the normalized total object count data (circles, squares, or triangles), which are globally fitted using a one-phase decay model (black line). Three biological repeats are shown for the normalized ThT aggregation data (blue lines), where each biological repeat is the mean of three technical replicates and error bars represent the standard deviation of the mean. e Representative DIC images acquired at the bottom of the well during FL α-syn incubation. Scale bars represent 25 μm. f Representative TEM image, with magnification, of an aliquot collected at the endpoint of a phase separation ThT aggregation assay. Scale bars represent 2 μm and 500 nm, respectively.

Article Snippet: The samples were incubated at 37 °C within the chamber of a Nikon ECLIPSE Ti2-E microscope (Nikon, Japan) and automated DIC microscopy imaging in solution was carried out 15 min from PLK addition using a 40x air objective.

Techniques: Sedimentation, Incubation, Imaging, Microscopy, Derivative Assay, Standard Deviation

Journal: Communications Chemistry

Article Title: Surface wetting is a key determinant of α-synuclein condensate maturation

doi: 10.1038/s42004-025-01764-z

Figure Lengend Snippet: a─c Representative DIC (top) and fluorescent (bottom) images of 60 µM FL α-syn incubated with 25 µM PLK in phase separation buffer at 37 °C. Samples were incubated in 8-well slides with either a glass ( a ), polymer ( b ) or BioInert ( c ) coverslip. Images were acquired at selected time points at the bottom surface of the well. Scale bars represent 25 µm. ThT fluorescence intensity quantification of the microscopy images acquired during the screening assay of glass ( d ), polymer ( e ) or BioInert ( f ) coverslips. Each data point is the average fluorescent intensity across the whole image, 3 images were quantified per time point. The mean intensity per time point is indicated by a solid black line, error bars represent the standard deviation of the mean.

Article Snippet: The samples were incubated at 37 °C within the chamber of a Nikon ECLIPSE Ti2-E microscope (Nikon, Japan) and automated DIC microscopy imaging in solution was carried out 15 min from PLK addition using a 40x air objective.

Techniques: Incubation, Polymer, Fluorescence, Microscopy, Screening Assay, Standard Deviation