clsm  (Advanced Microscopy Techniques)

Journal: Biofilm

Article Title: Inactivation of Cysteine Synthase CysK-A enhances flocculation, biofilm formation, and sensitivity to oxidative stress in Azospirillum brasilense Sp7

doi: 10.1016/j.bioflm.2025.100335

Figure Lengend Snippet: Three-dimensional (3D) reconstruction of A. brasilense biofilms by CLSM. Biofilms were stained to visualize some matrix components: red fluorescence indicates extracellular DNA (eDNA) stained with propidium iodide, and blue fluorescence indicates exopolysaccharides stained with Calcofluor. Upper panels: composite 3D images captured at 20x magnification, showing the distribution of both eDNA and EPS. Lower panels: Detailed 3D images of exopolysaccharides captured at 60x magnification. White arrows indicate structures resembling cyst-like cells within the biofilm of the cysK -A mutant strain. Scale bar = 25 μm. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: Confocal images were acquired using a Nikon C2+ CLSM (Nikon, Tokyo, Japan) equipped with a CFI Plan Apo Lambda 20 × objective and two helium–neon lasers for the excitation of the mCherry fluorophore at wavelengths of 540 nm and 650 nm and an argon laser for the excitation of radish autofluorescence at 488 nm.

Techniques: Staining, Fluorescence, Mutagenesis

Journal: Biofilm

Article Title: Inactivation of Cysteine Synthase CysK-A enhances flocculation, biofilm formation, and sensitivity to oxidative stress in Azospirillum brasilense Sp7

doi: 10.1016/j.bioflm.2025.100335

Figure Lengend Snippet: A. brasilense AR mutant forms thicker aggregates on radish roots than the wild-type strain. Representative 3D CLSM images of radish roots 7 days post-inoculation. ( A ) Wild-type (WT) A. brasilense Sp7 (harboring pMP2449-5) displays a typical rhizosphere colonization pattern. ( B ) cysK -A mutant ( A. brasilense AR, harboring pMP2449-5) forming visible, thicker, and denser aggregates (white arrows) compared to the WT. For both strains, bacteria are tagged with mCherry (red fluorescence), and the plant root autofluorescence is shown in green. Aggregate thickness was measured from the mCherry channel Z-stacks, revealing a depth of 42.5 μm for the WT and 50 μm for the AR mutant. Images were captured with a 20x objective, 2.0x digital zoom, and steps of 0.5 μm. Scale bar = 25 μm. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: Confocal images were acquired using a Nikon C2+ CLSM (Nikon, Tokyo, Japan) equipped with a CFI Plan Apo Lambda 20 × objective and two helium–neon lasers for the excitation of the mCherry fluorophore at wavelengths of 540 nm and 650 nm and an argon laser for the excitation of radish autofluorescence at 488 nm.

Techniques: Mutagenesis, Bacteria, Fluorescence

Journal: Biofilm

Article Title: Microplastics as active modulators of Escherichia coli biofilm characteristics and their implications on the development of antimicrobial resistance

doi: 10.1016/j.bioflm.2026.100355

Figure Lengend Snippet: Antibiotic susceptibility of biofilms exposed to ciprofloxacin for 24 h . (a) C-biofilms at 100, 200, and 250 μg/mL, (b) C-, G-, and MP-biofilm viability at 250 μg/mL, (c) Respective CLSM projected z-stacks of biofilms. (d) MP- and G-biofilms at varying concentrations of 300 and 350 μg/mL of ciprofloxacin. Green fluorescence denotes live E. coli cells and red indicates dead cells from exposure to ciprofloxacin. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: Stained biofilms were imaged with an inverted confocal laser scanning microscope (CLSM) (Olympus FV3000) with a 40 × objective.

Techniques: Fluorescence

Journal: Biofilm

Article Title: Microplastics as active modulators of Escherichia coli biofilm characteristics and their implications on the development of antimicrobial resistance

doi: 10.1016/j.bioflm.2026.100355

Figure Lengend Snippet: EPS composition and distribution in C-, MP- and G-biofilms (a) Total area of EPS signal in a cross-sectional CLSM capture (101250 μm 2 ), (b) percent composition of each constituent (eDNA, proteins, polysaccharides) found in EPS, (c) CLSM cross-sectional images of biofilms at the midpoint of the biofilm from the base (z = 0). Magenta, yellow, and cyan denote eDNA, proteins, and polysaccharides, respectively. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: Stained biofilms were imaged with an inverted confocal laser scanning microscope (CLSM) (Olympus FV3000) with a 40 × objective.

Techniques:

Journal: Journal of Functional Biomaterials

Article Title: Nanotopography-Mediated Mechanotransduction Enhances hBMSCs Adhesion on TiO 2 Nanotubes

doi: 10.3390/jfb17040200

Figure Lengend Snippet: Morphological characterization of hBMSCs cultured on different sample surfaces. ( A ) Representative SEM images with EDS of hBMSCs cultured on sample surfaces. Blue indicates the Ti element, representing the titanium substrate, while red indicates the C element, which mainly corresponds to the attached hBMSCs. Cells on the TS surface exhibited a rounded morphology, whereas cells on the anodized surface were flatter and more spread out. ( B ) Representative CLSM images of hBMSCs cultured on sample surfaces. The cytoskeletal organization was more pronounced in the anodized groups, and vinculin expression was most evident in the TNT10 group.

Article Snippet: The following supporting information can be downloaded at: https://www.mdpi.com/article/10.3390/jfb17040200/s1 , Figure S1: Schematic diagram of automatic identification of nanotubes based on Amira software; Figure S2: Surface elemental composition of the different samples determined via EDS; Figure S3: Representative CLSM images of hBMSCs cultured on sample surfaces (low-power view).

Techniques: Cell Culture, Expressing