Journal: Advanced Science

Article Title: Formation of Condition‐Dependent Alpha‐Synuclein Fibril Strain in Artificial Cerebrospinal Fluid

doi: 10.1002/advs.202505228

Figure Lengend Snippet: End products of aSyn aggregation recorded using AFM (A) and fibril morphology examples (B) that were formed in PB or aCSF. Cropped 2.5 µm x 2.5 µm images of the original 10 µm x 10 µm are shown in order to provide a clear view of different positions on the mica. The different structural polymorphs are readily visible at the displayed heatmaps for each example AFM image with fibril height and width traces. In aCSF3, * indicates analysed filament. Pitch and Max‐Min averages across the fibril are summarized in the adjacent table. A clear difference between PB type fibrils is visible, where PB3 does not contain expressed twist periodicity. Distinct fibril types observed in PB and aCSF using Cryo‐EM (C), along with their probability and crossover section of the 3D map. Each 2D class projection was analyzed using Fiji software to extract the full FWHM at the widest and narrowest positions for comparison. The cross‐section maps do not represent equal scaling.

Article Snippet: Cryo‐EM data were collected either on a Glacios transmission electron microscope (Fisher Scientific) operated at 200 kV camera or a Krios transmission electron microscope (Fisher Scientific) operated at 300 kV and equipped with Falcon 3EC/4i cameras.

Techniques: Cryo-EM Sample Prep, Software, Comparison

Journal: Advanced Science

Article Title: Formation of Condition‐Dependent Alpha‐Synuclein Fibril Strain in Artificial Cerebrospinal Fluid

doi: 10.1002/advs.202505228

Figure Lengend Snippet: Cryo‐EM model of aSyn fibrils formed in aCSF. Side view (A) and top view (B) of the reconstructed Cryo‐EM map. Single cross‐section of the atomic model (residues 1‐99) built in the density map (C). The salt bridge (D) is formed between K21 and E35 residues. Ligand density embedded inside the fibril core (E). The red and blue colors represent oxygen and nitrogen atoms, correspondingly, while orange/cyan display the carbon backbone. β‐sheet arrangements in the structure (F). Charge and hydrophobicity/hydrophilicity distributions (G) in the structure, where blue, green, and yellow colors represent very hydrophilic, hydrophilic, and hydrophobic amino acids, correspondingly.

Article Snippet: Cryo‐EM data were collected either on a Glacios transmission electron microscope (Fisher Scientific) operated at 200 kV camera or a Krios transmission electron microscope (Fisher Scientific) operated at 300 kV and equipped with Falcon 3EC/4i cameras.

Techniques: Cryo-EM Sample Prep

Journal: Advanced Science

Article Title: Formation of Condition‐Dependent Alpha‐Synuclein Fibril Strain in Artificial Cerebrospinal Fluid

doi: 10.1002/advs.202505228

Figure Lengend Snippet: aCSF fibril seeding process in aCSF and PB solutions (A) and the resulting aggregate composition formed in PB solution, characterized using Cryo‐EM (B). No significant difference between endpoint ThT fluorescence intensity was observed, while 2D cryo‐EM displays ≈1:9 ratio of two distinct fibril conformations. The resulting AFM images (C) of aCSF fibrils 10 min after resuspension in different aCSF solution compositions (indicated in the top left corner) and Cryo‐EM images (D) after incubating the samples for 24 h. The aCSF–HSA / aCSF indicates the fibrils resuspended in aCSF–HSA solution and then, after 10 min incubation, resuspended in aCSF. aCSF fibril imaging in PB solution on a mica using HS‐AFM (E).

Article Snippet: Cryo‐EM data were collected either on a Glacios transmission electron microscope (Fisher Scientific) operated at 200 kV camera or a Krios transmission electron microscope (Fisher Scientific) operated at 300 kV and equipped with Falcon 3EC/4i cameras.

Techniques: Cryo-EM Sample Prep, Fluorescence, Incubation, Imaging