Bacterial Stains General bacterial stains, fluorescent gram stains, and viability dyes Yeast and Fungi Stains General yeast stains, yeast viability dyes and kits, and organelle stains Viability PCR PMA PMAxx™ for fast, quantitative microbial viability by PCRAt 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 STAINSFor 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 StainsOften 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. BactoView Live Fluorescent Bacterial Stains Nucleic acid binding dyes Green and red options Useful for fluorescence microscopy or flow cytometry RedDot 1 Far-Red DNA Dye Nucleic acid binding dye Far-red fluorescence Useful for fluorescence microscopy DMAO, 2 mM in DMSO Green nucleic acid binding dye Useful for fluorescence microscopy or flow cytometry Hoechst 33342, 10 mg/mL in H2O Blue nucleic acid binding dye Useful for fluorescence microscopy or flow cytometry Thiazole Orange Nucleic acid binding dye Green fluorescence Useful for fluorescence microscopy or flow cytometry CellBrite Fix Membrane Stains Fixable membrane dyes Green, red and far-red options Useful for fluorescence microscopyE. coli stained with BactoView™ Live Green (green, DNA) and SynaptoRed™ C2 (magenta, membrane). Bacillus subtilis stained with CellBrite™ Fix 555, fixed in paraformaldehyde, and imaged the next day.Fluorescent Gram StainsA 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. Bacterial Viability and Gram Stain KitCF 488A WGA stains gram-positive cells greenEthD-III stains dead cells redDAPI stains all cells blueA co-culture of E. coli and Staphylococcus epidermidis 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.Co-culture of E. coli and Staphylococcus epidermidis stained with CF®594 WGA  and DAPI. Gram-positive Staphylococcus show 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. CF®488A WGA stains gram-positive Micrococcus green, EthDIII stains dead cells red, and DAPI stains all cells blue.Bacterial Viability StainsBiotium 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). Live-or-Dye™ Fixable Viability Staining KitsFixable dead cell stainsAvailable in 8 fluorescent colorsUseful for flow cytometry or microscopy Live-or-Dye NucFix™ Red Staining Kit Fixable, DNA-binding dead cell stainWorks in gram-negative bacteria only Stains dead cells red RedDot 2 Far-Red DNA DyeNuclear-specific dead cell stainStains dead cells with far-red fluorescence Viability/Cytotoxicity Assay KitGreen fluorescent DMAO stains DNA in all bacteriaEthidium Homodimer III (EthD-III) stains dead cells red Bacterial Viability and Gram Stain KitCF 488A WGA stains gram-positive cells greenEthDIII stains dead cells redDAPI stains alll cells blue 5-Cyano-2,3-ditolyl tetrazolium chloride (CTC)Substrate for measuring bacterial respiratory activityHealthy cells reduce CTC into an insoluble red fluorescent formazan productA 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 blueA 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 topYEAST STAINS AND KITSFluorescent 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 StainsBiotium 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). ViaVac Red/GreenYeast-specific vitality dyestains the vacuole red in metabolically active cells Yeast Live-or-Dye™ Fixable Live/Dead Staining KitThiazole Orange stains all cells greenLive-or-Dye™ 568/583 stains dead cells redFixable staining Yeast Viability Staining KitCF® Dye-labeled Concanavalin A (ConA) to label the cell walls of all cells Live-or-Dye™ viability dye to label dead cellsChoose blue/green, green/red, or red/far-red color combinations Yeast Vitality Staining KitCalcofluor White labels all cell walls blueViaVac™ Red/Green stains the vacuole red in metabolically active cellsS. cerevisiae 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).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).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 YeastA 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. MitoView™ DyesMitochondrial dyes in five colors: MitoView™ 633 is mitochondrial membrane potential-dependent LysoView™ 540 633Label vacuoles in live yeast cells2 LysoView dyes stain the yeast vacuole LysoView 540 LysoView 633 LipidSpot™ 488Label vacuoles in live yeast cellsStains intracellular membrane structures Calcofluor WhiteBlue fluorescent chitin stain stains cell wallsStrongly stains bud scars CellBrite™ FixStains the cell membraneAvailable in 3 dye colorsStaining is fixable MemBrite™ FixStains the cell peripheryAvailable in 8 dye colorsStaining is fixable CF Dye Concanavalin A (Con A) ConjugatesCon A lectin binds to a-mannopyranosyl and a-glucopyranosy in the cell wall of yeast and fungiAvailable in 10 colors CF Dye Wheat Germ Agglutinin (WGA) ConjugatesWGA lectin binds sialic acid and N-acetylglucosamineStrongly labels bud scarsAvailable in 13 colors Thiazole OrangeGreen nucleic acid binding dyeCan be used as an all-cell yeast stainStaining in yeast is nuclear in unfixed cells and cytoplasmic after fixation NucSpot® 470 Nuclear StainGreen nucleic acid binding dyeStains the nucleus of dead or fixed and permeabilized yeast Phalloidin conjugatesPhalloidin stains F-actin cables and patches in fixed and permeabilized yeastAvailable in 20 colors and biotinS. 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. cerevisiae stained separately with MemBrite™ Fix 488/515, 568/580, and 640/660, then combined and imaged by confocal microscopy.Mitochondrial staining of S. cerevisiae with MitoView™ 633. Imaged on a Zeiss confocal microscope.Fixed and permeabilized S. cerevisiae stained with CF®568-conjugated phalloidinHeat-killed S. cerevisiae stained with NucSpot®470 Nuclear Stain.Back to topVIABILITY 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 PCRPMA 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 PMAxx , 20 mM in H2O40069 Next-generation v-PCR dye Gives the best live/dead discrimination PMA Dye40013 Original v-PCR dye developed at Biotium Validated in hundreds of publications and a wide variety of organismsViability PCR Starter KitsThe 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 . Viability PCR Starter Kit with PMA and Enhancer31076PMAGram-negative bacteria Viability PCR Starter Kit with PMAxx and Enhancer31076-XPMAxx Gram-negative bacteriaStrain-Specific Viability PCR KitsStrain-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 KitsPMA Enhancer for Gram-Negative BacteriaPMA 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. PMA Enhancer for Gram Negative Bacteria, 5X Solution31038 Improves live/dead discrimination in gram-negative strainsPhotoactivation DevicesThe 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. PMA-Lite LED Photolysis DeviceE90002 Photoactivation device, holds 16 microcentrifuge tubes Glo-Plate Blue LED IlluminatorE90004 Photoactivation device for plates or larger tubes From: 155Sizes: 5 x 1 mgCatalog #: 29015, 29016, 29017, 29018, 29019, 29020, 29058, 29074, 29075, - 29080View allHide From: 68Sizes: 1 mg, 5 x 1 mgCatalog #: 29021, 29022, 29023, 29024, 29025, 29026, 29027, 29028, 29029, 29059, 29064, 29073, 29021-1, 29022-1, 29023-1, 29024-1, 29025-1, 29026-1, 29027-1, 29028-1, 29029-1, 29059-1, 29064-1, 29077-1, 29077, 29076-1, - 29076View allHide Biotium implements a Quality System, certified by QAS according to Standard QAS ISO 9001:2015. Certificate No. US3564