Microbiology is the study of microscopic organisms, such as bacteria, viruses, archaea, fungi and protozoa. This discipline includes fundamental research on biochemistry, physiology, cell biology, ecology, evolution and clinical aspects of microorganisms including the host response to these agents. We are pleased to offer the products needed to a microbiologist to carry his work.

We have a range  of infra red sterilisers, bead busters for your microbiology work.

 

Features

  • HM-300A
  • GBS-3000A
  • Bioprep 24
    In addition we also carry a range of microbiological stains, kits and dyes.
    General bacterial stains, fluorescent gram stains, and viability dyes
    General yeast stains, yeast viability dyes and kits, and organelle stains
    PMA & PMAxx™ for fast, quantitative microbial viability by PCR

    At Biotium, our specialty is creating fluorescent dyes that simplify and improve your scientific experiments. For Microbiology, we offer a wide range of fluorescent dyes and assays kits to stain bacteria, yeast and fungi, including dyes to assess cell viability and bacterial gram stain type. We have also developed a complete toolkit of reagents for viability PCR, a revolutionary method for determining microbial viability using qPCR.

    Also visit our Cellular Stains Table for a comprehensive list of cellular stains with their ability to stain various cell types.


    BACTERIAL STAINS

    For bacteria, we offer fluorescent dyes to stain live cells, dead cells, and gram+ cells. Combining these dyes in multi-color microscopy or flow cytometry experiments allows several parameters to be assessed at one time.

    Bacterial Live Cell Stains

    Often it can be useful to visualize or detect all bacteria present in a sample, regardless of species or metabolic state. For this purpose, we offer a selection of dyes that generally stain all bacteria, both gram-positive and gram-negative. The dyes are available in various colors and localizations:

    • BactoView™ Live, RedDot™1, DMAO, Thiazole Orange, and Hoechst are cell membrane permeant nucleic acid binding dyes.
    • SynaptoGreen™, SynaptoRed™, and CellBrite™ Fix are all cell membrane/cell periphery stains. CellBrite™ Fix dyes are a set of fixable membrane dyes available in multiple colors.
    E. coli stained with BactoView™ Live Green (stains DNA) and SynaptoRed™ (stains the membrane)
    Bacillus subtilis stained with CellBrite™ Fix 555, fixed in paraformaldehyde, and imaged the next day.

    Fluorescent Gram Stains

    A traditional colorimetric gram staining protocol is laborious, requiring up to 5 different solutions to stain cells, including toxic alcohol and acetone to decolorize the sample, and precise timing. In contrast, Wheat Germ Agglutinin (WGA) has been shown to bind specifically to N-acetylglucosamine in the peptidoglycan layer of the cell wall of gram-positive bacteria, and can be labeled with a number of different fluorescent dyes for one-step fluorescent cell labeling. We offer fluorescent WGA conjugates in 13 color options, as well as combination gram stain/viability kits and DNA dyes.

    Product Name Features
    Wheat Germ Agglutinin (WGA) CF® Dye conjugates •   Fast and sensitive gram stains
    •   Available in 13 colors
    Live Bacterial Gram Stain Kit •   CF®594 WGA stains gram-positive cells red
    •   DAPI stains all cells blue
    Bacterial Viability and Gram Stain Kit •   CF®488A WGA stains gram-positive cells green
    •   EthD-III stains dead cells red
    •   DAPI stains all cells blue

     

    Microbiology Stain - E. coli and Salmonella co-culture stained with DAPI and CF633-WGA
    A co-culture of E. coli and Staphylococcus stained with DAPI (blue) and CF®633 WGA (red). Spherically-shaped gram-positive Staphylococcus  stain with WGA, while rod-shaped gram-negative E. coli do not stain. DAPI stains all cells blue.
    E. coli and Staphylococcus stained with CF594-WGA and DAPI
    Co-culture of E. coli and Staphylococcus stained with CF®594 WGA  and DAPI. Gram-positive Staphylococcusshow blue DAPI staining and red cell surface staining with WGA, while gram-negative E. coli show only DAPI staining.
    Heat-killed Pseudomonas and Micrococcus stained with the Bacterial Viability and Gram Stain Kit
    Heat-killed Pseudomonas and Micrococcus stained with the Bacterial Viability and Gram Stain Kit. CF®488A WGA stains gram-positive Micrococcus green, EthDIII stains dead cells red, and DAPI stains all cells blue.

    Bacterial Viability Stains

    Biotium offers a selection of different fluorescent viability dyes and kits, to allow for the differentiation of live and dead cells by microscopy or flow cytometry (see the Viability PCR section below for detection of viable bacteria by qPCR).

    Product Name Features
    Live-or-Dye™ Fixable Viability Staining Kits •   Fixable dead cell stains
    •   Available in 8 fluorescent colors
    •   Useful for flow cytometry or microscopy
    Live-or-Dye NucFix™ Red Staining Kit •   Fixable, DNA-binding dead cell stain
    •   Works in gram-negative bacteria only
    •   Stains dead cells red
    RedDot™2 Far-Red DNA Dye •   Nuclear-specific dead cell stain
    •   Stains dead cells with far-red fluorescence
    Ethidium Homodimer III (EthD-III) •   Nuclear-specific dead cell stain
    •   Stains dead cells red
    Viability/Cytotoxicity Assay Kit •   Green fluorescent DMAO stains DNA in all bacteria
    •   Ethidium Homodimer III (EthD-III) stains dead cells red
    Bacterial Viability and Gram Stain Kit •   CF®488A WGA stains gram-positive cells green
    •   EthDIII stains dead cells red
    •   DAPI stains alll cells blue
    5-Cyano-2,3-ditolyl tetrazolium chloride (CTC) •   Substrate for measuring bacterial respiratory activity
    •   Healthy cells reduce CTC into an insoluble red fluorescent formazan product

     

    A mixture of live and dead E. coli stained with Live-or-Dye™ 568/583 and DAPI. Live-or-Dye™ 568/583 stains dead cells red, and DAPI stains all cells blue
    A mixture of live and dead E. coli stained with the Viability/Cytotoxicity Kit. DMAO stains all cells green and EthD-III stains dead cells red.

    Back to top


    YEAST STAINS AND KITS

    Fluorescent dyes and kits to stain yeast cells, including viability dyes, combination staining kits, and organelle stains. Combining these dyes in multi-color microscopy or flow cytometry experiments allows maximum experimental versatility.

    Yeast Viability Stains

    Biotium offers a selection of different fluorescent viability dyes and kits, to allow for the differentiation of live and dead yeast cells by microscopy or flow cytometry (see the Viability PCR section below for detection of viable yeast by qPCR).

    Product Name Features
    Live-or-Dye™ Fixable Viability Staining Kits •   Fixable dead cell stains
    •   Choice of 8 fluorescent colors
    Propidium Iodide •   Red dead cell stain
    •   Not fixable
    Ethidium Homodimer III (EthD-III) •   Far-red dead cell stain
    •   Not fixable
    RedDot™2 Far-Red Nuclear Stain •   Far-red, nuclear dead cell stain
    •   Not fixable
    NucSpot® 470 •   Green, nuclear dead cell stain
    •   Not fixable
    ViaVac™ Red/Green •   Yeast-specific vitality dye
    •   stains the vacuole red in metabolically active cells
    Yeast Live-or-Dye™ Fixable Live/Dead Staining Kit •   Thiazole Orange stains all cells green
    •   Live-or-Dye™ 568/583 stains dead cells red
    •   Fixable staining
    Yeast Viability Staining Kit •   CF® Dye-labeled Concanavalin A (ConA) to label the cell walls of all cells
    •   Live-or-Dye™ viability dye to label dead cells
    •   Choose blue/green, green/red, or red/far-red color combinations
    Yeast Vitality Staining Kit •   Calcofluor White labels all cell walls blue
    •   ViaVac™ Red/Green stains the vacuole red in metabolically active cells

     

    S. cereviciae stained with the Yeast Vitality Staining Kit: Calcofluor White stains all cell walls blue and ViaVac™ stains the vacuoles of metabolically active cells red and the cytoplasm of all cells green (not shown).
    Yeast Live-or-Dye Fixable Live/Dead Staining Kit
    S. cerevisiae stained with the Yeast Live-or-Dye™ Fixable Live/Dead Staining Kit: Thiazole Orange to stain all cells (green) and Live-or-Dye™ 568/583 to stain dead cells (red, appears yellow in overlay).
    Yeast Viability Staining Kit
    S. cerevisiae stained with the Yeast Viability Staining Kit: CF®640R ConA stains cell walls of all cells with far-red fluorescence (pseudocolored blue in image), while Live-or-Dye™ 568/583 stains dead cells red.

    Organelle Stains for Yeast

    A number of our organelle and cellular stains can be used to stain yeast cells. We also offer stains for the fungal cell wall and bud scars of budding yeast.

    Product Name Features
    MitoView™ Dyes •   Mitochondrial dyes in five colors:
    •   MitoView™ 633 is mitochondrial membrane potential-dependent
    LysoView™ 540 & 633 •   Label vacuoles in live yeast cells
    •   2 LysoView dyes stain the yeast vacuole LysoView™ 540 & LysoView™ 633
    LipidSpot™ 488 •   Label vacuoles in live yeast cells
    •   Stains intracellular membrane structures
    Calcofluor White •   Blue fluorescent chitin stain stains cell walls
    •   Strongly stains bud scars
    CellBrite™ Fix •   Stains the cell membrane
    •   Available in 3 dye colors
    •   Staining is fixable
    MemBrite™ Fix •   Stains the cell periphery
    •   Available in 8 dye colors
    •   Staining is fixable
    CF® Dye Concanavalin A (Con A) Conjugates •   Con A lectin binds to a-mannopyranosyl and a-glucopyranosy in the cell wall of yeast and fungi
    •   Available in 10 colors
    CF® Dye Wheat Germ Agglutinin (WGA) Conjugates •   WGA lectin binds sialic acid and N-acetylglucosamine
    •   Strongly labels bud scars
    •   Available in 13 colors
    Thiazole Orange •   Green nucleic acid binding dye
    •   Can be used as an all-cell yeast stain
    •   Staining in yeast is nuclear in unfixed cells and cytoplasmic after fixation
    NucSpot® 470 Nuclear Stain •   Green nucleic acid binding dye
    •   Stains the nucleus of dead or fixed and permeabilized yeast
    ViaFluor® CFSE •   Can be used as an all-cell cytoplasmic stain in yeast
    Phalloidin conjugates •   Phalloidin stains F-actin cables and patches in fixed and permeabilized yeast
    •   Available in 20 colors and biotin

     

    S. cerevisiae stained with CF®488A WGA (green) to stain bud scars and CF®594 ConA (red) to stain the cell wall.
    S. cerevisiae stained with LysoView™ 633 to visualize the vacuoles. Imaged on a Zeiss confocal microscope.
    S. cereviciae stained separately with MemBrite™ Fix 488/515, 568/580, and 640/660. Combined and imaged.
    Mitochondrial staining of S. cerevisiae with MitoView™ 633. Imaged on a Zeiss confocal microscope.
    Fixed and permeabilized S. cerevisiae stained with CF®568-conjugated phalloidin
    Heat-killed S. cerevisiae stained with NucSpot®470 Nuclear Stain.

    Back to top


    VIABILITY PCR WITH PMA & PMAXX™

    Biotium is the inventor of the original viability PCR dye, PMA, and the new and improved viability PCR dye PMAxx™. We have also developed a large range of products and kits for viability PCR. To learn more about this technology and all of the associated products, please visit the Viability PCR Technology Page.


    Introduction to viability PCR (v-PCR)

    Viability PCR is a powerful technology for the sensitive and rapid detection of viable microorganisms. Unlike time-consuming culturing methods, qPCR is a fast and sensitive method of detection. However, normal qPCR does not distinguish between live and dead cells. With v-PCR using PMAxx™ or PMA, you get the speed, sensitivity and specificity of PCR, plus quantifiable viability. And because no culturing is required, you can even detect viable but not culturable (VBNC) bacteria.The v-PCR technology can be applied not only to bacteria but to other organisms like yeast, viruses, eukaryotes, and archaea.

    How does v-PCR work?

    The infographic below shows the basic mechanism of v-PCR. PMAxx™ and PMA are photoreactive dyes with high affinity for DNA. The dyes intercalate into dsDNA and form a covalent linkage upon exposure to intense visible light. The reaction inhibits PCR amplification of modified DNA templates by a combination of removal of modified DNA during purification and inhibition of template amplification by DNA polymerases. Because the dyes are cell membrane-impermeable, when a sample containing both live and dead bacteria is treated with dye, only dead bacteria with compromised cell membranes are susceptible to DNA modification. In a qPCR reaction, dead cell DNA will show delayed amplification and higher Ct than live cells.

    Viability PCR dyes like PMAxx™ or PMA are membrane-impermeant, which makes them dead cell specific. Once inside of a dead cell, they bind to DNA. Exposure to intense visible light renders the dyes reactive, and causes them to covalently attach to the DNA. This DNA modification prevents amplification in subsequent PCR reactions.

    PMA & PMAxx™ Dyes for Viability PCR

    PMA is the original dye developed at Biotium for v-PCR. It has been validated in a wide variety of bacterial strains, as well as yeast and viruses (see PMA References). However, while PMA is generally effective at differentiating between live and dead bacteria by qPCR, it does not completely eliminate PCR products from dead cell DNA. This could potentially give false positive results.

    PMAxx™ was designed by Biotium scientists to be a superior alternative to PMA. PMAxx™ is much more effective at eliminating PCR amplification of dead cell DNA, and therefore provides the best discrimination between live and dead bacteria.

    Download the PMA™ and PMAxx™ flyer to learn more.

    Live or dead bacteria treated with either PMA or PMAxx™ prior to qPCR. Dead bacteria treated with either PMA or PMAxx™ show delays in amplification compared to either live cells, or untreated dead cells. Bacteria treated with PMAxx™ show a larger delay than those treated with PMA. Therefore PMAxx™ is better able to differentiate live vs dead bacteria by viability PCR.

    PMA & PMAxx™ for Viability PCR

    Product Name Catalog No. Features
    PMAxx™, 20 mM in H2O 40069 •    Next-generation vPCR dye
    •    Gives the best live/dead discrimination
    PMA Dye 40013 •    Original vPCR dye developed at Biotium
    •    Validated in hundreds of publications and a wide variety of organisms
    PMA Dye, 20 mM in H2O 40019

    Other Viability PCR Reagents

    Viability PCR Starter Kits

    The customizable Viability PCR Starter Kits contain the materials that you need for selective detection of viable cells using either PMA or PMAxx™ viability dye and qPCR. You also have the choice of selecting a kit with or without the Enhancer for Gram Negative Bacteria (not to be used with other cell types). The user will need to supply their own primers to amplify their species of interest .


    Strain-Specific Viability PCR Kits

    Strain-specific v-PCR kits are designed for selective detection of viable bacteria from a specific strain using PMAxx or PMA dye and real-time PCR. The kits contain PMA dye OR PMAxx™ dye, Forget-Me-Not™ EvaGreen® qPCR Master Mix, and validated PCR primers for detection of selected strains of bacteria.

    Strain-Specific Bacterial Viability PCR Kits

    Bacteria Strain Gene name Kit with PMA (catalog #) Kit with PMAxx™ (catalog #)
    Salmonella enterica invA 31033 31033-X
    Mycobacterium tuberculosis groEL2 31034 N/A
    Staphylococcus aureus nuc 31035 N/A
    MRSA mecA 31036 N/A
    E. coli O157:H7 Z3276 31037 31037-X
    E. coli uidA 31050 31050-X
    Listeria monocytogenes hly 31051 31051-X
    Legionella pneumophila mip 31053 N/A

    PMA Enhancer for Gram-Negative Bacteria

    PMA Enhancer for Gram Negative Bacteria was designed to improve discrimination between live and dead gram-negative bacteria during viability PCR. PMA Enhancer is provided as a 5X solution, and is added to a sample before the addition of viability dye.

    Product Name Catalog No. Features
    PMA Enhancer for Gram Negative Bacteria, 5X Solution 31038 •    Improves live/dead discrimination in gram-negative strains

    Photoactivation Devices

    The PMA-Lite™ LED photolysis device allows for time- and temperature-controlled light treatment of PMAxx- or PMA-treated samples in microcentrifuge tubes.

    The Glo-Plate™ Blue LED Illuminator is designed for even light treatment of PMAxx™- or PMA-treated samples in plates or larger tubes.

    Product Name Catalog No. Features
    PMA-Lite™ LED Photolysis Device E90002 •    Photoactivation device, holds 16 microcentrifuge tubes
    Glo-Plate™ Blue LED Illuminator E90004 •    Photoactivation device for plates or larger tubes

    Back to top