cell imaging

Mitochondrial-derived vesicles retain membrane potential and contain a functional ATP synthase

by NANOSENSORS
15 March 202415 March 2024
Science & Technology

Figure 2 B from Reut Hazan et al. 2023 “Mitochondrial‐derived vesicles retain membrane potential and contain a functional ATP synthase”: Characterization of MDVs from wild‐type mitochondria B. Atomic force microscopy. Representative AFM image and a 3D AFM image of one representative vesicle (WT), adsorbed on a mica modified with Mg2+ and imaged under PBS. Please refer to the full article cited below to view the full figure. NANOSENSORSTM uniqprobeTM qp-BioAC-CI AFM probes with rounded AFM tips designed for cell imaging were used for the characterization by atomic force microscopy (AFM) in QI mode.

Vesicular transport is a means of communication. * While cells can communicate with each other via secretion of extracellular vesicles, less is known regarding organelle‐to… Read More »Mitochondrial-derived vesicles retain membrane potential and contain a functional ATP synthase

Male-female communication enhances release of extracellular vesicles leading to high fertility in Drosophila

by NANOSENSORS
28 October 202215 March 2023
Science & Technology

Fig. 5 from “Male-female communication enhances release of extracellular vesicles leading to high fertility in Drosophila” by Javier Arturo Sanchez-Lopez et al.: The spermatheca releases EVs with specific characteristics. a–e The spermathecae (SSC > CD63-GFP) were cultured ex vivo to characterize the particles that were released to the spent media: a size distribution of CD63-GFP-positive particles (n = 25 spermathecae from 15 flies) using single-particle tracking of the Nanoimager. b Characterization of CD63-GFP particles by ExoView (100 spermathecae from 50-60 flies) using anti-GFP or anti-CD63. c Representative dSTORM image of EVs found in the spent media of SSC expressing CD63-GFP (n = 25 spermathecae from 15 flies), stained with AlexaFluor555 conjugated anti-CD63 primary antibody. The image shows CD63-GFP-positive EVs and single CD63 AlexaFluor555 molecules the EV’s surface. Scale bars = 1 µm and insets = 20 nm. d–e The morphology of EVs in the spent media was observed by e negative staining in STEM; scale bar = 200 nm; and by d Cryo-TEM; Scale bar = 100 nm (see also Supplementary Fig. 5a-c). f–i The spent media of ex vivo cultured spermathecae were analyzed for the presence of EVs: f Representative AFM scans of SSC-EVs isolated by acoustic sorting (n = 100 spermathecae from 70 flies), showing from left to right: height (nm), adhesion (pN) and Young’s modulus images (MPa). Scale bars = 100 nm (see Supplementary Fig. 5d–g). g–i Single-particle tracking in the nanoimager. g CellMaskTM-positive particle size distribution and concentration profiles of EVs in the spent media of spermathecae incubated in media alone (No ecdy) or with ecdysone (Ecdy) (see also Supplementary Movie 5 for a time lapse of spermathecae end apparatus and endosomal activity post-ecdysone stimulation and methods, ex vivo spermatheca culture section); media with only CellMaskTM stain served as a control for the formation of dye aggregates. h Particle concentration and i mean particle diameter (nm) from g; Box plots are the measurements of particles from four frames of spent media from 25 spermathecae; boxes represent maximum, median and minimum values with outliers; one-way ANOVA, with nonparametric Wilcoxon multiple comparison post-hoc test; *p

The female reproductive tract (female-RT) must decipher the repertoire of molecular cues received from the male during copulation in order to activate and coordinate tract… Read More »Male-female communication enhances release of extracellular vesicles leading to high fertility in Drosophila

Simultaneous co-localized super-resolution fluorescence microscopy and atomic force microscopy: combined SIM and AFM platform for the life sciences

by NANOSENSORS
9 June 202015 March 2023
Science & Technology

Figure 4 a and b from Ana I. Gómez-Varela et al “Simultaneous co-localized super-resolution fluorescence microscopy and atomic force microscopy: combined SIM and AFM platform for the life sciences : Simultaneous SR-SIM/AFM acquisition. The AFM measurements were carried out on fixed U2OS cells in medium/buffer with (a) and without N-SIM illumination (b). For convenience and enhanced feature/noise contrast, both AFM topography images in the SR-SIM/AFM overlays are displayed with an edge detection algorithm using a pixel difference operator in X. The topography images from Petri dish surface on three positions (labelled in the figures) were planefit (1st order polynomial function) to compensate for tilts in the sample surface, and subjected to surface roughness analysis Please have a look at the full article to view the full figure. https://rdcu.be/b4Iot

Correlating data from different microscopy techniques holds the potential to discover new facets of signalling events in cellular biology.* In the article “Simultaneous co-localized super-resolution… Read More »Simultaneous co-localized super-resolution fluorescence microscopy and atomic force microscopy: combined SIM and AFM platform for the life sciences

Substrate properties modulate cell membrane roughness by way of actin filaments

by NANOSENSORS
4 January 201915 March 2023
Science & Technology

Supplementary Figure S1 from Chao-Hung Chang et al. “Substrate properties modulate cell membrane roughness by way of actin filaments”: Images of membrane topography determined by atomic force microscopy (AFM). MEFs were seeded on the polymer coverslip-bottom μ-dishes coated with 0 or 10 μg/ml FN for 6 hours for the measurement of membrane roughness by AFM. The regions marked by the white squares in the bright-field images are displayed in the membrane topography. Scale bar, 10 μm. NANOSENSORS uniqprobe qp-SCONT AFM probes(long cantilever length 125 um, spring constant 0.01 N/m) were used.

“Cell membrane roughness has been proposed as a sensitive feature to reflect cellular physiological conditions”* In the article “Substrate properties modulate cell membrane roughness by… Read More »Substrate properties modulate cell membrane roughness by way of actin filaments