RESUMO
Biofilms are structured communities of bacteria that are held together by an extracellular matrix consisting of protein and exopolysaccharide. Biofilms often have a limited lifespan, disassembling as nutrients become exhausted and waste products accumulate. D-amino acids were previously identified as a self-produced factor that mediates biofilm disassembly by causing the release of the protein component of the matrix in Bacillus subtilis. Here we report that B. subtilis produces an additional biofilm-disassembly factor, norspermidine. Dynamic light scattering and scanning electron microscopy experiments indicated that norspermidine interacts directly and specifically with exopolysaccharide. D-amino acids and norspermidine acted together to break down existing biofilms and mutants blocked in the production of both factors formed long-lived biofilms. Norspermidine, but not closely related polyamines, prevented biofilm formation by B. subtilis, Escherichia coli, and Staphylococcus aureus.
Assuntos
Bacillus subtilis/fisiologia , Biofilmes , Polissacarídeos Bacterianos/metabolismo , Espermidina/análogos & derivados , Aminoácidos/biossíntese , Aminoácidos/metabolismo , Bacillus subtilis/genética , Escherichia coli/fisiologia , Mutação , Poliaminas/metabolismo , Espermidina/biossíntese , Espermidina/metabolismo , Staphylococcus aureus/fisiologiaRESUMO
Modern molecular microbiology elucidates the organizational principles of bacterial biofilms via detailed examination of the interplay between signaling and gene regulation. A complementary biophysical approach studies the mesoscopic dependencies at the cellular and multicellular levels with a distinct focus on intercellular forces and mechanical properties of whole biofilms. Here, motivated by recent advances in biofilm research and in other, seemingly unrelated fields of biology and physics, we propose a perspective that links the biofilm, a dynamic multicellular organism, with the physical processes occurring in the extracellular milieu. Using Bacillus subtilis as an illustrative model organism, we specifically demonstrate how such a rationale explains biofilm architecture, differentiation, communication, and stress responses such as desiccation tolerance, metabolism, and physiology across multiple scales-from matrix proteins and polysaccharides to macroscopic wrinkles and water-filled channels.
Assuntos
Bacillus subtilis , Biofilmes , Biofilmes/crescimento & desenvolvimento , Bacillus subtilis/fisiologia , Bacillus subtilis/metabolismo , Regulação Bacteriana da Expressão Gênica , Proteínas de Bactérias/metabolismo , Proteínas de Bactérias/genéticaRESUMO
Microbiology began as a unified science using the principles of chemistry to understand living systems. The unified view quickly split into the subdisciplines of medical microbiology, molecular biology, and environmental microbiology. The advent of a universal phylogeny and culture-independent approaches has helped tear down the boundaries separating the subdisciplines. The vision for the future is that the study of the fundamental roles of microbes in ecology and evolution will lead to an integrated biology with no boundary between microbiology and macrobiology.
Assuntos
Ecologia , Biologia Molecular , Microbiologia , FilogeniaRESUMO
While scientific research should be carried out objectively, the choices of questions asked and approaches taken are deeply personal and subjective. I urge individuals to pursue questions they love and to periodically scrutinize the reasons (the philosophies) that drive that love. As a case study, I scrutinize the "whys" behind some of the scientific questions I pursued during my career.
Assuntos
Escolha da Profissão , Humanos , História do Século XXIRESUMO
The Introduction of antibiotics into the clinical use in the middle of the 20th century had a profound impact on modern medicine and human wellbeing. The contribution of these wonder molecules to public health and science is hard to overestimate. Much research has informed our understanding of antibiotic mechanisms of action and resistance at inhibitory concentrations in the lab and in the clinic. Antibiotics, however, are not a human invention as most of them are either natural products produced by soil microorganisms or semisynthetic derivatives of natural products. Because we use antibiotics to inhibit the bacterial growth, it is generally assumed that growth inhibition is also their primary ecological function in the environment. Nevertheless, multiple studies point to diverse nonlethal effects that are exhibited at lower levels of antibiotics. Here we review accumulating evidence of antibiosis and of alternative functions of antibiotics exhibited at subinhibitory concentrations. We also speculate on how these effects might alter phenotypes, fitness, and community composition of microbes in the context of the environment and suggest directions for future research.
Assuntos
Anti-Infecciosos/metabolismo , Anti-Infecciosos/farmacologia , Fenômenos Ecológicos e Ambientais/efeitos dos fármacos , Antibacterianos/farmacologia , Antibiose/genética , Antibiose/fisiologia , Humanos , Microbiologia do SoloRESUMO
Bacterial pathogens have evolved strategies that enable them to invade tissues and spread within the host. Enterococcus faecalis is a leading cause of local and disseminated multidrug-resistant hospital infections, but the molecular mechanisms used by this non-motile bacterium to penetrate surfaces and translocate through tissues remain largely unexplored. Here we present experimental evidence indicating that E. faecalis generates exopolysaccharides containing ß-1,6-linked poly-N-acetylglucosamine (polyGlcNAc) as a mechanism to successfully penetrate semisolid surfaces and translocate through human epithelial cell monolayers. Genetic screening and molecular analyses of mutant strains identified glnA, rpiA and epaX as genes critically required for optimal E. faecalis penetration and translocation. Mechanistically, GlnA and RpiA cooperated to generate uridine diphosphate N-acetylglucosamine (UDP-GlcNAc) that was utilized by EpaX to synthesize polyGlcNAc-containing polymers. Notably, exogenous supplementation with polymeric N-acetylglucosamine (PNAG) restored surface penetration by E. faecalis mutants devoid of EpaX. Our study uncovers an unexpected mechanism whereby the RpiA-GlnA-EpaX metabolic axis enables production of polyGlcNAc-containing polysaccharides that endow E. faecalis with the ability to penetrate surfaces. Hence, targeting carbohydrate metabolism or inhibiting biosynthesis of polyGlcNAc-containing exopolymers may represent a new strategy to more effectively confront enterococcal infections in the clinic.
Assuntos
Enterococcus faecalis/metabolismo , Matriz Extracelular de Substâncias Poliméricas/fisiologia , Polissacarídeos Bacterianos/fisiologia , Proteínas de Bactérias , Enterococcus faecalis/patogenicidade , Matriz Extracelular de Substâncias Poliméricas/metabolismo , Infecções por Bactérias Gram-Positivas , Humanos , Polissacarídeos Bacterianos/metabolismoRESUMO
As part of this Special Issue of International Microbiology celebrating the 75th anniversary of the founding of the Spanish Society for Microbiology (SEM), Guest Editor Rafael Giraldo invited us to contribute an opinion article on the topic of 75 years of joint scientific ventures between Latin American microbiologists and Spanish microbiologists. Since the creation of SEM in 1945 (Pérez Prieto, NoticiaSEM 98:1, 2016) Latin American microbiologists have been participants, both as individuals and as members of the national associations that are currently integrated into the Latin American Association for Microbiology (ALAM). Thus, the histories of Spanish and Latin American microbiology (Chica and Skinner, Int Microbiol 13:159-164, 2010; Chica, Int Microbiol 11:221-225, 2008) have been closely linked over the last 75 years. In order to provide our perspective on the topic, we decided to provide answers to three questions:What are key aspects of the history of Spanish and Latin American Microbiology interactions? What have been some of our personal experiences in which these interactions shaped our careers? What is our outlook for the future of such interactions?
Assuntos
Microbiologia , Humanos , América LatinaRESUMO
The rapid emergence of antibiotic-resistant pathogenic bacteria has accelerated the search for new antibiotics. Many clinically used antibacterials were discovered through culturing a single microbial species under nutrient-rich conditions, but in the environment, bacteria constantly encounter poor nutrient conditions and interact with neighboring microbial species. In an effort to recapitulate this environment, we generated a nine-strain actinomycete community and used 16S rDNA sequencing to deconvolute the stochastic production of antimicrobial activity that was not observed from any of the axenic cultures. We subsequently simplified the community to just two strains and identified Amycolatopsis sp. AA4 as the producing strain and Streptomyces coelicolor M145 as an inducing strain. Bioassay-guided isolation identified amycomicin (AMY), a highly modified fatty acid containing an epoxide isonitrile warhead as a potent and specific inhibitor of Staphylococcus aureus Amycomicin targets an essential enzyme (FabH) in fatty acid biosynthesis and reduces S. aureus infection in a mouse skin-infection model. The discovery of AMY demonstrates the utility of screening complex communities against specific targets to discover small-molecule antibiotics.
Assuntos
Antraquinonas/farmacologia , Antibacterianos/farmacologia , Streptomyces coelicolor/crescimento & desenvolvimento , Antraquinonas/química , Antibacterianos/química , DNA Bacteriano/genética , DNA Bacteriano/metabolismo , DNA Ribossômico/genética , DNA Ribossômico/metabolismo , Testes de Sensibilidade Microbiana/métodos , RNA Ribossômico 16S/genética , RNA Ribossômico 16S/metabolismo , Streptomyces coelicolor/genéticaRESUMO
Bacillus subtilis forms organized multicellular communities known as biofilms wherein the individual cells are held together by a self-produced extracellular matrix. The environmental signals that promote matrix synthesis remain largely unknown. We discovered that one such signal is impaired respiration. Specifically, high oxygen levels suppressed synthesis of the extracellular matrix. In contrast, low oxygen levels, in the absence of an alternative electron acceptor, led to increased matrix production. The response to impaired respiration was blocked in a mutant lacking cytochromes caa3 and bc and markedly reduced in a mutant lacking kinase KinB. Mass spectrometry of proteins associated with KinB showed that the kinase was in a complex with multiple components of the aerobic respiratory chain. We propose that KinB is activated via a redox switch involving interaction of its second transmembrane segment with one or more cytochromes under conditions of reduced electron transport. In addition, a second kinase (KinA) contributes to the response to impaired respiration. Evidence suggests that KinA is activated by a decrease in the nicotinamide adenine dinucleotide (NAD(+))/NADH ratio via binding of NAD(+) to the kinase in a PAS domain A-dependent manner. Thus, B. subtilis switches from a unicellular to a multicellular state by two pathways that independently respond to conditions of impaired respiration.
Assuntos
Bacillus subtilis/fisiologia , Proteínas de Bactérias/metabolismo , Biofilmes , Citocromos/metabolismo , Proteínas Quinases/metabolismo , Bacillus subtilis/efeitos dos fármacos , Bacillus subtilis/enzimologia , Bacillus subtilis/genética , Matriz Extracelular/efeitos dos fármacos , Matriz Extracelular/metabolismo , Regulação Bacteriana da Expressão Gênica , Histidina Quinase , Ferro/farmacologia , Mutação , NAD/metabolismo , Oxigênio/metabolismo , Ligação Proteica , Oligoelementos/farmacologiaRESUMO
Plant-associated microbes are important for the growth and health of their hosts. As a result of numerous prior studies, we know that host genotypes and abiotic factors influence the composition of plant microbiomes. However, the high complexity of these communities challenges detailed studies to define experimentally the mechanisms underlying the dynamics of community assembly and the beneficial effects of such microbiomes on plant hosts. In this work, from the distinctive microbiota assembled by maize roots, through host-mediated selection, we obtained a greatly simplified synthetic bacterial community consisting of seven strains (Enterobacter cloacae, Stenotrophomonas maltophilia, Ochrobactrum pituitosum, Herbaspirillum frisingense, Pseudomonas putida, Curtobacterium pusillum, and Chryseobacterium indologenes) representing three of the four most dominant phyla found in maize roots. By using a selective culture-dependent method to track the abundance of each strain, we investigated the role that each plays in community assembly on roots of axenic maize seedlings. Only the removal of E. cloacae led to the complete loss of the community, and C. pusillum took over. This result suggests that E. cloacae plays the role of keystone species in this model ecosystem. In planta and in vitro, this model community inhibited the phytopathogenic fungus Fusarium verticillioides, indicating a clear benefit to the host. Thus, combined with the selective culture-dependent quantification method, our synthetic seven-species community representing the root microbiome has the potential to serve as a useful system to explore how bacterial interspecies interactions affect root microbiome assembly and to dissect the beneficial effects of the root microbiota on hosts under laboratory conditions in the future.
Assuntos
Bactérias/isolamento & purificação , Zea mays/microbiologia , Bactérias/classificação , Bactérias/genética , Microbiota , Filogenia , Raízes de Plantas/microbiologia , Microbiologia do SoloRESUMO
The organization of cells, emerging from cell-cell interactions, can give rise to collective properties. These properties are adaptive when together cells can face environmental challenges that they separately cannot. One particular challenge that is important for microorganisms is migration. In this study, we show how flagellum-independent migration is driven by the division of labor of two cell types that appear during Bacillus subtilis sliding motility. Cell collectives organize themselves into bundles (called "van Gogh bundles") of tightly aligned cell chains that form filamentous loops at the colony edge. We show, by time-course microscopy, that these loops migrate by pushing themselves away from the colony. The formation of van Gogh bundles depends critically on the synergistic interaction of surfactin-producing and matrix-producing cells. We propose that surfactin-producing cells reduce the friction between cells and their substrate, thereby facilitating matrix-producing cells to form bundles. The folding properties of these bundles determine the rate of colony expansion. Our study illustrates how the simple organization of cells within a community can yield a strong ecological advantage. This is a key factor underlying the diverse origins of multicellularity.
Assuntos
Bacillus subtilis/fisiologia , Diferenciação Celular , Movimento , Bacillus subtilis/química , Percepção de QuorumRESUMO
Bacteria form dense surface-associated communities known as biofilms that are central to their persistence and how they affect us. Biofilm formation is commonly viewed as a cooperative enterprise, where strains and species work together for a common goal. Here we explore an alternative model: biofilm formation is a response to ecological competition. We co-cultured a diverse collection of natural isolates of the opportunistic pathogen Pseudomonas aeruginosa and studied the effect on biofilm formation. We show that strain mixing reliably increases biofilm formation compared to unmixed conditions. Importantly, strain mixing leads to strong competition: one strain dominates and largely excludes the other from the biofilm. Furthermore, we show that pyocins, narrow-spectrum antibiotics made by other P. aeruginosa strains, can stimulate biofilm formation by increasing the attachment of cells. Side-by-side comparisons using microfluidic assays suggest that the increase in biofilm occurs due to a general response to cellular damage: a comparable biofilm response occurs for pyocins that disrupt membranes as for commercial antibiotics that damage DNA, inhibit protein synthesis or transcription. Our data show that bacteria increase biofilm formation in response to ecological competition that is detected by antibiotic stress. This is inconsistent with the idea that sub-lethal concentrations of antibiotics are cooperative signals that coordinate microbial communities, as is often concluded. Instead, our work is consistent with competition sensing where low-levels of antibiotics are used to detect and respond to the competing genotypes that produce them.
Assuntos
Antibiose , Biofilmes/crescimento & desenvolvimento , Pseudomonas aeruginosa/crescimento & desenvolvimento , Piocinas/farmacologia , Antibacterianos , Biofilmes/efeitos dos fármacos , Técnicas de Cocultura , MicrofluídicaRESUMO
Kin discrimination, broadly defined as differential treatment of conspecifics according to their relatedness, could help biological systems direct cooperative behavior toward their relatives. Here we investigated the ability of the soil bacterium Bacillus subtilis to discriminate kin from nonkin in the context of swarming, a cooperative multicellular behavior. We tested a collection of sympatric conspecifics from soil in pairwise combinations and found that despite their history of coexistence, the vast majority formed distinct boundaries when the swarms met. Some swarms did merge, and most interestingly, this behavior was only seen in the most highly related strain pairs. Overall the swarm interaction phenotype strongly correlated with phylogenetic relatedness, indicative of kin discrimination. Using a subset of strains, we examined cocolonization patterns on plant roots. Pairs of kin strains were able to cocolonize roots and formed a mixed-strain biofilm. In contrast, inoculating roots with pairs of nonkin strains resulted in biofilms consisting primarily of one strain, suggestive of an antagonistic interaction among nonkin strains. This study firmly establishes kin discrimination in a bacterial multicellular setting and suggests its potential effect on ecological interactions.
Assuntos
Bacillus subtilis/fisiologia , Biofilmes , Interações Microbianas/fisiologia , Raízes de Plantas/microbiologia , Sequência de Aminoácidos , Sequência de Bases , Análise por Conglomerados , Primers do DNA/genética , Modelos Genéticos , Dados de Sequência Molecular , Filogenia , Análise de Sequência de DNA , Especificidade da EspécieRESUMO
The membranes of eukaryotic cells harbor microdomains known as lipid rafts that contain a variety of signaling and transport proteins. Here we show that bacterial membranes contain microdomains functionally similar to those of eukaryotic cells. These membrane microdomains from diverse bacteria harbor homologs of Flotillin-1, a eukaryotic protein found exclusively in lipid rafts, along with proteins involved in signaling and transport. Inhibition of lipid raft formation through the action of zaragozic acid--a known inhibitor of squalene synthases--impaired biofilm formation and protein secretion but not cell viability. The orchestration of physiological processes in microdomains may be a more widespread feature of membranes than previously appreciated.
Assuntos
Bactérias/metabolismo , Proteínas de Bactérias/metabolismo , Microdomínios da Membrana/metabolismo , Bacillus subtilis/enzimologia , Bacillus subtilis/metabolismo , Bactérias/enzimologia , Biofilmes/efeitos dos fármacos , Inibidores Enzimáticos/metabolismo , Farnesil-Difosfato Farnesiltransferase/metabolismo , Proteínas de Membrana/metabolismo , Transporte Proteico , Transdução de Sinais/efeitos dos fármacos , Desidrogenase do Álcool de Açúcar/metabolismoRESUMO
Growing cells of Bacillus subtilis are a bistable mixture of individual motile cells in which genes for daughter cell separation and motility are ON, and chains of sessile cells in which these genes are OFF. How this ON/OFF switch is controlled has been mysterious. Here we report that a complex of the SinR and SlrR proteins binds to and represses genes involved in cell separation and motility. We also report that SinR and SlrR constitute a double-negative feedback loop in which SinR represses the gene for SlrR (slrR), and, by binding to (titrating) SinR, SlrR prevents SinR from repressing slrR. Thus, SlrR indirectly derepresses its own gene, creating a self-reinforcing loop. Finally, we show that, once activated, the loop remains locked in a high SlrR state in which cell separation and motility genes are OFF for extended periods of time. SinR and SlrR constitute an epigenetic switch for controlling genes involved in cell separation and motility.
Assuntos
Bacillus subtilis/crescimento & desenvolvimento , Bacillus subtilis/genética , Proteínas de Bactérias/genética , Epigênese Genética , Biofilmes/crescimento & desenvolvimento , Divisão Celular/genética , Retroalimentação Fisiológica , Regulação Fúngica da Expressão Gênica , Modelos Moleculares , N-Acetil-Muramil-L-Alanina Amidase/genética , Mutação Puntual , Regiões Promotoras Genéticas/genética , Ligação Proteica , Estrutura Terciária de ProteínaRESUMO
Cellular differentiation is triggered by extracellular signals that cause target cells to adopt a particular fate. Differentiation in bacteria typically involves autocrine signaling in which all cells in the population produce and respond to the same signal. Here we present evidence for paracrine signaling in bacterial populations-some cells produce a signal to which only certain target cells respond. Biofilm formation in Bacillus involves two centrally important signaling molecules, ComX and surfactin. ComX triggers the production of surfactin. In turn, surfactin causes a subpopulation of cells to produce an extracellular matrix. Cells that produced surfactin were themselves unable to respond to it. Likewise, once surfactin-responsive cells commenced matrix production, they no longer responded to ComX and could not become surfactin producers. Insensitivity to ComX was the consequence of the extracellular matrix as mutant cells unable to make matrix responded to both ComX and surfactin. Our results demonstrate that extracellular signaling was unidirectional, with one subpopulation producing a signal and a different subpopulation responding to it. Paracrine signaling in a bacterial population ensures the maintenance, over generations, of particular cell types even in the presence of molecules that would otherwise cause those cells to differentiate into other cell types.
Assuntos
Bacillus subtilis/fisiologia , Comunicação Parácrina , Bacillus subtilis/crescimento & desenvolvimento , Bacillus subtilis/metabolismo , Proteínas de Bactérias/metabolismo , Matriz Extracelular/metabolismo , Regulação Bacteriana da Expressão Gênica , Lipopeptídeos/metabolismo , Peptídeos Cíclicos/metabolismoRESUMO
The microalga Emiliania huxleyi produces alkenone lipids that are important proxies for estimating past sea surface temperatures. Field calibrations of this proxy are robust but highly variable results are obtained in culture. Here, we present results suggesting that algal-bacterial interactions may be responsible for some of this variability. Co-cultures of E. huxleyi and the bacterium Phaeobacter inhibens resulted in a 2.5-fold decrease in algal alkenone-containing lipid bodies. In addition levels of unsaturated alkenones increase in co-cultures. These changes result in an increase in the reconstructed growth temperature of up to 2°C relative to axenic algal cultures.
Assuntos
Haptófitas/metabolismo , Haptófitas/microbiologia , Lipídeos/química , Rhodobacteraceae/fisiologia , Haptófitas/química , Metabolismo dos Lipídeos , Microalgas/metabolismo , Microalgas/microbiologia , TemperaturaRESUMO
Bacillus subtilis is a plant-beneficial Gram-positive bacterium widely used as a biofertilizer. However, relatively little is known regarding the molecular processes underlying this bacterium's ability to colonize roots. In contrast, much is known about how this bacterium forms matrix-enclosed multicellular communities (biofilms) in vitro. Here, we show that, when B. subtilis colonizes Arabidopsis thaliana roots it forms biofilms that depend on the same matrix genes required in vitro. B. subtilis biofilm formation was triggered by certain plant polysaccharides. These polysaccharides served as a signal for biofilm formation transduced via the kinases controlling the phosphorylation state of the master regulator Spo0A. In addition, plant polysaccharides are used as a source of sugars for the synthesis of the matrix exopolysaccharide. The bacterium's response to plant polysaccharides was observed across several different strains of the species, some of which are known to have beneficial effects on plants. These observations provide evidence that biofilm genes are crucial for Arabidopsis root colonization by B. subtilis and provide insights into how matrix synthesis may be triggered by this plant.
Assuntos
Bacillus subtilis/fisiologia , Biofilmes/crescimento & desenvolvimento , Matriz Extracelular/metabolismo , Raízes de Plantas/microbiologia , Polissacarídeos/metabolismo , Análise de Variância , Arabidopsis , Bacillus subtilis/metabolismo , Proteínas de Bactérias/metabolismo , Carbono/metabolismo , Primers do DNA/genética , Citometria de Fluxo , Galactanos , Processamento de Imagem Assistida por Computador , Microscopia de Fluorescência , Raízes de Plantas/metabolismo , Plasmídeos/genética , Polissacarídeos/química , Especificidade da Espécie , Fatores de Transcrição/metabolismoRESUMO
Biofilms, surface-bound communities of microbes, are economically and medically important due to their pathogenic and obstructive properties. Among the numerous strategies to prevent bacterial adhesion and subsequent biofilm formation, surface topography was recently proposed as a highly nonspecific method that does not rely on small-molecule antibacterial compounds, which promote resistance. Here, we provide a detailed investigation of how the introduction of submicrometer crevices to a surface affects attachment of Escherichia coli. These crevices reduce substrate surface area available to the cell body but increase overall surface area. We have found that, during the first 2 h, adhesion to topographic surfaces is significantly reduced compared with flat controls, but this behavior abruptly reverses to significantly increased adhesion at longer exposures. We show that this reversal coincides with bacterially induced wetting transitions and that flagellar filaments aid in adhesion to these wetted topographic surfaces. We demonstrate that flagella are able to reach into crevices, access additional surface area, and produce a dense, fibrous network. Mutants lacking flagella show comparatively reduced adhesion. By varying substrate crevice sizes, we determine the conditions under which having flagella is most advantageous for adhesion. These findings strongly indicate that, in addition to their role in swimming motility, flagella are involved in attachment and can furthermore act as structural elements, enabling bacteria to overcome unfavorable surface topographies. This work contributes insights for the future design of antifouling surfaces and for improved understanding of bacterial behavior in native, structured environments.
Assuntos
Aderência Bacteriana/fisiologia , Biofilmes , Escherichia coli/fisiologia , Flagelos/fisiologia , Escherichia coli/ultraestrutura , Flagelos/ultraestrutura , Microscopia de Força Atômica , Microscopia Confocal , Microscopia Eletrônica de Varredura , Propriedades de Superfície , Fatores de TempoRESUMO
Fluorescence microscopy is a method commonly used to examine individual differences between bacterial cells, yet many studies still lack a quantitative analysis of fluorescence microscopy data. Here we introduce some simple tools that microbiologists can use to analyze and compare their microscopy images. We show how image data can be converted to distribution data. These data can be subjected to a cluster analysis that makes it possible to objectively compare microscopy images. The distribution data can further be analyzed using distribution fitting. We illustrate our methods by scrutinizing two independently acquired data sets, each containing microscopy images of a doubly labeled Bacillus subtilis strain. For the first data set, we examined the expression of srfA and tapA, two genes which are expressed in surfactin-producing and matrix-producing cells, respectively. For the second data set, we examined the expression of eps and tapA; these genes are expressed in matrix-producing cells. We show that srfA is expressed by all cells in the population, a finding which contrasts with a previously reported bimodal distribution of srfA expression. In addition, we show that eps and tapA do not always have the same expression profiles, despite being expressed in the same cell type: both operons are expressed in cell chains, while single cells mainly express eps. These findings exemplify that the quantification and comparison of microscopy data can yield insights that otherwise would go unnoticed.