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Phase contrast and fluorescence microscopic images of Bacillales spores stained with APBT and safranin O. cells of Bacillus subtilis (A) , B. licheniformis (B) , Niallia circulans (C) , <t>Brevibacillus</t> brevis (D) , Lysinibacillus sphaericus (E) , and Paenibacillus polymyxa (F) were suspended in a mixture of 2-(4-aminophenyl) benzothiazole (APBT) and safranin O and observed using phase contrast and fluorescence microscopy. From left to right: phase contrast images, APBT fluorescence, green fluorescence of safranin O, red fluorescence of safranin O, and merged images. Blue represents APBT fluorescence, while green and red indicate the green and red fluorescence of safranin O, respectively. Arrows indicate vegetative cells (VCs), mother cells (MCs), and forespores (FSs), and arrowheads indicate mature spores (MSs) in phase-contrast microscopy images. The scale bar represents 5 μm.
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Phase contrast and fluorescence microscopic images of Bacillales spores stained with APBT and safranin O. cells of Bacillus subtilis (A) , B. licheniformis (B) , Niallia circulans (C) , <t>Brevibacillus</t> brevis (D) , Lysinibacillus sphaericus (E) , and Paenibacillus polymyxa (F) were suspended in a mixture of 2-(4-aminophenyl) benzothiazole (APBT) and safranin O and observed using phase contrast and fluorescence microscopy. From left to right: phase contrast images, APBT fluorescence, green fluorescence of safranin O, red fluorescence of safranin O, and merged images. Blue represents APBT fluorescence, while green and red indicate the green and red fluorescence of safranin O, respectively. Arrows indicate vegetative cells (VCs), mother cells (MCs), and forespores (FSs), and arrowheads indicate mature spores (MSs) in phase-contrast microscopy images. The scale bar represents 5 μm.
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Phase contrast and fluorescence microscopic images of Bacillales spores stained with APBT and safranin O. cells of Bacillus subtilis (A) , B. licheniformis (B) , Niallia circulans (C) , <t>Brevibacillus</t> brevis (D) , Lysinibacillus sphaericus (E) , and Paenibacillus polymyxa (F) were suspended in a mixture of 2-(4-aminophenyl) benzothiazole (APBT) and safranin O and observed using phase contrast and fluorescence microscopy. From left to right: phase contrast images, APBT fluorescence, green fluorescence of safranin O, red fluorescence of safranin O, and merged images. Blue represents APBT fluorescence, while green and red indicate the green and red fluorescence of safranin O, respectively. Arrows indicate vegetative cells (VCs), mother cells (MCs), and forespores (FSs), and arrowheads indicate mature spores (MSs) in phase-contrast microscopy images. The scale bar represents 5 μm.
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Phase contrast and fluorescence microscopic images of Bacillales spores stained with APBT and safranin O. cells of Bacillus subtilis (A) , B. licheniformis (B) , Niallia circulans (C) , <t>Brevibacillus</t> brevis (D) , Lysinibacillus sphaericus (E) , and Paenibacillus polymyxa (F) were suspended in a mixture of 2-(4-aminophenyl) benzothiazole (APBT) and safranin O and observed using phase contrast and fluorescence microscopy. From left to right: phase contrast images, APBT fluorescence, green fluorescence of safranin O, red fluorescence of safranin O, and merged images. Blue represents APBT fluorescence, while green and red indicate the green and red fluorescence of safranin O, respectively. Arrows indicate vegetative cells (VCs), mother cells (MCs), and forespores (FSs), and arrowheads indicate mature spores (MSs) in phase-contrast microscopy images. The scale bar represents 5 μm.
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Phase contrast and fluorescence microscopic images of Bacillales spores stained with APBT and safranin O. cells of Bacillus subtilis (A) , B. licheniformis (B) , Niallia circulans (C) , Brevibacillus brevis (D) , Lysinibacillus sphaericus (E) , and Paenibacillus polymyxa (F) were suspended in a mixture of 2-(4-aminophenyl) benzothiazole (APBT) and safranin O and observed using phase contrast and fluorescence microscopy. From left to right: phase contrast images, APBT fluorescence, green fluorescence of safranin O, red fluorescence of safranin O, and merged images. Blue represents APBT fluorescence, while green and red indicate the green and red fluorescence of safranin O, respectively. Arrows indicate vegetative cells (VCs), mother cells (MCs), and forespores (FSs), and arrowheads indicate mature spores (MSs) in phase-contrast microscopy images. The scale bar represents 5 μm.

Journal: Frontiers in Microbiology

Article Title: Combination of fluorescent reagents with 2-(4-aminophenyl) benzothiazole and safranin O was useful for analysis of spore structure, indicating the diversity of Bacillales species spores

doi: 10.3389/fmicb.2025.1603957

Figure Lengend Snippet: Phase contrast and fluorescence microscopic images of Bacillales spores stained with APBT and safranin O. cells of Bacillus subtilis (A) , B. licheniformis (B) , Niallia circulans (C) , Brevibacillus brevis (D) , Lysinibacillus sphaericus (E) , and Paenibacillus polymyxa (F) were suspended in a mixture of 2-(4-aminophenyl) benzothiazole (APBT) and safranin O and observed using phase contrast and fluorescence microscopy. From left to right: phase contrast images, APBT fluorescence, green fluorescence of safranin O, red fluorescence of safranin O, and merged images. Blue represents APBT fluorescence, while green and red indicate the green and red fluorescence of safranin O, respectively. Arrows indicate vegetative cells (VCs), mother cells (MCs), and forespores (FSs), and arrowheads indicate mature spores (MSs) in phase-contrast microscopy images. The scale bar represents 5 μm.

Article Snippet: In addition to B. subtilis , the following Bacillales spore-forming bacteria were used: Niallia circulans NBRC 13629, B. licheniformis ATCC 14580, Brevibacillus brevis NBRC 100599, Lysinibacillus sphaericus NBRC 3526, and Paenibacillus polymyxa NBRC15309.

Techniques: Fluorescence, Staining, Microscopy