Fig. 3: Volumetric imaging and dynamic tracking of intracellular processes using VLM. | Nature Communications

Fig. 3: Volumetric imaging and dynamic tracking of intracellular processes using VLM.

From: Volumetric localization microscopy with deep learning

Fig. 3: Volumetric imaging and dynamic tracking of intracellular processes using VLM.The alternative text for this image may have been generated using AI.

a Wide-field and 3D VLM images of AF647-labeled clathrin-coated pits (CCPs) in U-2 OS cells. b Cross-sectional projection of the corresponding boxed region in (a), revealing sparse 3D distribution of CCPs. c Schematic of CCP formation in three stages. Created in BioRender. Gao, Z. (2025) https://BioRender.com/m95z857. d Wide-field (left) and 3D VLM (right) images of three representative CCPs, as indicated by arrows in (a), at distinct formation stages with cross-sectional profiles (insets). e Raw elemental light-field images of MitoTracker-labeled mitochondria in HeLa cells. f 3D VLM images of the boxed region in (e) at time point t = 0.0 s, showing depth-color-coded emitters overlaid on corresponding light-field reconstructed images using Richardson-Lucy deconvolution. g Time-resolved MitoTracker dynamic changes through rolling-window analysis during high-power laser exposure at time points t = 2.4, 3.0, and 3.6 s. h Raw elemental light-field images of lysosomes (Ls, red) and peroxisomes (Pr, green) in live HeLa cells. i Two-color 3D reconstructed image by V-Net (t = 0.0 s) overlaid on the corresponding bright-field image. j Time-projected trajectories over 8 s of lysosomes and peroxisomes in the boxed region in (i). k VLM (top) and light-field reconstructed (bottom) images of two adjacent peroxisomes, as indicated in (i) at time points t = 1.12 and 6.43 s. l 3D trajectories of the two peroxisomes in (k) tracked over 8 s. m Zoomed-in images of two lysosomes (boxed in i), resolved by VLM at time points t = 0.5, 1.5, and 3.5 s, overlaid on corresponding light-field reconstructed images. n 3D trajectories of the two lysosomes in (m) tracked over 8 s. o Comparative diffusion coefficients of lysosomes (n = 8, mean = 7.09 × 10−3 μm2 s−1) and peroxisomes (n = 8, mean = 0.65 × 10−3 μm2 s−1). Box plots show the median (central line), lower and upper quartiles (box), and the minimum and maximum values within 1.5× the interquartile range (whiskers). Scale bars: 2 μm (a, f), 100 nm (b, d), 20 μm (e, h, i), 500 nm (g, j, k, m).

Back to article page