0

EbbaBiolight

EbbaBiolight Mix&Try - Test Kit for Getting Started

EbbaBiolight Mix&Try contains 10 µl each optotracer molecule available in the EbbaBiolight series. 

Following fluorescent tracer molecules are included in the EbbaBiolight Mix&Try  kit:

  • EbbaBiolight 480 (Excitation: 405-458 nm, Emission: 470–550 nm, Emax = 480 nm)
  • EbbaBiolight 520 (Excitation: 405-488 nm, Emission: 500-600 nm, Emax = 520 nm)
  • EbbaBiolight 540 (Excitation: 430-500 nm, Emission: 530-600 nm, Emax = 540 nm)
  • EbbaBiolight 630 (Excitation: 458-514 nm, Emission: 600-650 nm, Emax = 630 nm)
  • EbbaBiolight 680 (Excitation: 530-565 nm, Emission: 600-800 nm, Emax = 680 nm)

Using all these different options will allow you to select the best EbbaBiolight for your experiment. Please contact us for custom options.

contact us for custom options.

EbbaBiolight 680 - red/far-red optotracer molecule for probing and detecting bacterial and biofilm.

With exceptionally high signal-to-noise ratio and spectral properties that are clearly distinguishable from biological autofluorescence, we especially recommend EbbaBiolight 680 for tracking of biofilm in live cultures.

Use any type of fluorescence microscopy equipment (confocal microscope, spectroscope or spectrophotometer) to detect and visualize the fluorescence of EbbaBiolight 680 when bound to a target in the bacterial membrane or extracellular matrix. EbbaBiolight 680 has an optical spectrum that allows custom settings within the detection range of 600-800 nm and an excitation range of 530-565 nm. Emission can be detected at 680 nm using the standard PI (Propodium Iodide), mCherry or Cy3.5 filter set and excitation is achieved using the 561 nm laser line.

EbbaBiolight 680 is available as 1x 50 µl and 2x 50 µl, with a volume discount. Please contact us for custom options.

contact us for custom options.

EbbaBiolight 630 - orange optotracer molecule for probing and detecting bacterial and biofilm.

Use any type of fluorescence microscopy equipment (confocal microscope, spectroscope or spectrophotometer) to detect and visualize the fluorescence of EbbaBiolight 630 when bound to a target in the bacterial membrane or extracellular matrix. EbbaBiolight 630 has an optical spectrum that allows custom settings within the detection range of 600-650 nm and an excitation range of 458-514 nm. Emission can be detected at 630 nm using the standard PI (Propodium Iodide), Cy3, TxRed, mCherry or Cy3.5 filter set and excitation is achieved using the 488 or 514 nm laser line.

EbbaBiolight 630 is available as 1x 50 µl and 2x 50 µl, with a volume discount. Please contact us for custom options.

contact us for custom options.

EbbaBiolight 540 - green/yellow optotracer molecule for probing and detecting bacterial and biofilm.

Use any type of fluorescence microscopy equipment (confocal microscope, spectroscope or spectrophotometer) to detect and visualize the fluorescence of EbbaBiolight 540 when bound to a target in the bacterial membrane or extracellular matrix. EbbaBiolight 540 has an optical spectrum that allows custom settings within the detection range of 530-600 nm and excitation range of 430-500 nm. Emission can be detected at 540 nm using the standard FITC, GFP or YFP filter set when excited using the 440 nm laser line.

EbbaBiolight 540 is available as 1x 50 µl and 2x 50 µl, with a volume discount. Please contact us for custom options.

contact us for custom options.

EbbaBiolight 520 - green optotracer molecule for probing and detecting bacterial and biofilm.

Use any type of fluorescence microscopy equipment (confocal microscope, spectroscope or spectrophotometer) to detect and visualize the fluorescence of EbbaBiolight 520 when bound to a target in the bacterial membrane or extracellular matrix. EbbaBiolight 520 has an optical spectrum that allows custom settings within the detection range of 500-600 nm and excitation range of 405-488 nm. Emission can be detected at 520 nm using the standard FITC or GFP filter set when excited using the 458 or 488 nm laser line.

EbbaBiolight 520 is available as 1x 50 µl and 2x 50 µl, with a volume discount. Please contact us for custom options.

contact us for custom options.

EbbaBiolight 480 - blue optotracer molecule for probing and detecting bacterial and biofilm.

Use any type of fluorescence microscopy equipment (confocal microscope, spectroscope or spectrophotometer) to detect and visualize the fluorescence of EbbaBiolight 480 when bound to a target in the bacterial membrane or extracellular matrix. EbbaBiolight 480 has an optical spectrum that allows custom settings within the detection range of 470-550 nm and excitation range of 405-458 nm. Emission can be detected at 480 nm using the standard DAPI filter set filter set when excited using the 405 nm laser line.

EbbaBiolight 480 is available as 1x 50 µl and 2x 50 µl, with a volume discount. Please contact us for custom options.

contact us for custom options.

Optotracing

Optotracing is an innovative visualization method developed by scientists for scientists. With optotracing – using fluorescent tracer molecules – Ebba Biotech provides an innovative method for expanding areas of research. The optotracing technology has been developed by a scientific expert team of researchers at Karolinska Institutet, Linköping University and the Royal Institute of Technology (KTH) in Sweden and is a result of many years of cutting-edge nanoscience and organic chemistry research.

We supply and deliver the fluorescent tracer (optotracer) molecules for detection of bacteria and bacterial biofilm in the EbbaBiolight product series. The EbbaBiolight optotracers are a tool to search and detect bacteria & biofilm in hospitals and microbiology laboratories, as well as in industry with high demands to detect bacterial contamination and biofilm formation.

Optotracing with EbbaBiolight

EbbaBiolight are optotracers for detection of bacteria & components of bacterial biofilm.

EbbaBiolight molecules allow direct visualisation of bacteria or bacterial biofilm, without using antibodies or any toxic chemicals. The molecules become highly fluorescent when bound to a target, are non-toxic, and do not interfere with bacterial growth or biofilm formation at recommended concentrations so EbbaBiolight can be added to live cultures to track bacterial growth or biofilm formation in real-time using spectrophotometric analysis. Further, EbbaBiolight are photo- and thermostable and allow for easy handling in any application.

We offer the EbbaBiolight optotracers in five different variants that might be screened for applications in various bacterial species and strains. All molecules are available in aliquots of 50 µl or 100 µl (delivered as 2x 50 µl). We also offer an EbbaBiolight Mix&Try kit, that contains 10 µl of each of the five optical tracer molecules from the EbbaBiolight series.

Quick Facts

EbbaBiolight optotracer molecules are:

  • provided as 1 mg/ml solution in volumes of 50 µl and 100 µl
  • should be diluted 1000-fold before use
  • non-toxic
  • photo- and thermostable

Quickstart Guide

Depending on the bacterial species or strain, EbbaBiolight optotracer molecules will bind to components in the bacterial biofilm. We recommend to follow this Quickstart Guide when using any of the EbbaBiolight molecules for the first time.

  • Dilute EbbaBiolight 1:1000 in growth medium (pH 7.4).
  • Dilute a bacterial over-night culture 1:100 with growth medium containing EbbaBiolight.
  • Fill (at least) two wells of a 96-well microtiter plate with 100 μl of the diluted over-night culture containing EbbaBiolight.
  • Prevent drying by filling unused wells with sterile water and seal the plate with a cover or adhesive.
  • Incubate the plate at optimal growth temperature and measure absorbance as well as fluorescence at regular intervals during a period of at least 48h.
  • Take samples (10 µl) during exponential- and stationary phase and pipet onto a microscope slide for imaging. Cover with a glass coverslip and seal the edges to prevent drying. Image at recommended excitation/emission to detect fluorescent bacteria or biofilm.

Applications for EbbaBiolight 680

EbbaBiolight 680 is our optotracer for biofilm visualization. EbbaBiolight 680 binds to cellulose and amyloid components in the biofilm of Salmonella enterica, serovars S. Enteritidis and Salmonella Typhimurium.

Storage

  • Store EbbaBiolight at 4°C.
  • Use the opened vial within 12 month.

Note

  • EbbaBiolight is for research use only.
  • EbbaBiolight is not for diagnostic use or use in humans.
  • EbbaBiolight is not for resale.

Download Info

Protocol I: Staining of surface biofilm using EbbaBiolight 680

This protocol describes how to grow biofilm at the air-liquid interface using the inclined-coverslip method and how to detect the air-surface biofilm using EbbaBiolight 680. As the optotracer does not influence biofilm formation when used in recommended concentrations, it can be present in growing cultures. It is also possible to grow the biofilm using the described method without EbbaBiolight 680, and then use EbbaBiolight 680 to visualize the grown biofilm as described in Protocol II. If necessary for imaging, we recommend light fixation in ice-cold ethanol, but fixation in 4% paraformaldehyde works as well. We have tested this procedure with Salmonella Enteritidis and Salmonella Typhimurium strains. For these strains, we have not observed staining of intracellular or membrane components.

Solutions and Reagents:

EbbaBiolight 680 is provided as 1000-fold concentrated solution. The following common reagents are required (not supplied):

  • Sterile glass coverslips (24x24 mm)
  • 6-well plate with cover or adhesive seal
  • Growth medium
  • Phosphate buffered saline (PBS) pH 7.4
  • Ethanol, 95% (-20°C) or 4% PFA
  • Mounting medium

Assay Procedure:

  • Place two sterile glass coverslips opposite to each other and inclined towards the walls of the wells in a 6-well plate.
  • Prepare your bacterial culture at appropriate bacterial density. Then, add EbbaBiolight 680 (1:1000) and mix gently before pipetting 5-7 ml of the solution into each well fitted with glass coverslips.
  • Seal the plate with a cover or adhesive seal and incubate at a temperature of choice for 24-48 h.
  • Remove the glass coverslips from each well. Wipe the backside clean and fix biofilm formed at the air-liquid interface on the glass coverslip by immersing the coverslip in cold ethanol for 4 min or PFA for 30 min.
  • Wash 3 x 1 min in PBS (Optional).
  • Mount the samples using mounting medium and seal the coverslip onto the slide to prevent drying.

Fluorescence Microscopy:

  • EbbaBiolight 680 is excited at 561 nm (standard laser line) and emission is detected using a standard PI (Propodium Iodide), mCherry or Cy3.5 filter set. Optional: An excitation range of 530-565 nm and a detection range of 600-800 nm may be applied depending on available laser lines and filter sets.

Download Protocol

Protocol II: Optotracing of biofilm in tissue sections using EbbaBiolight 680

This protocol describes how to stain for the biofilm markers curli and cellulose in tissue sections. EbbaBiolight can be used to stain tissue sections prepared by the most common techniques like paraffin embedding and freezing. We recommend fixation in ice-cold ethanol, but fixation in 4% paraformaldehyde works as well. We have tested this procedure with Salmonella Enteritidis and Salmonella Typhimurium strains. For these strains we have not observed staining of intracellular or membrane components. As EbbaBiolight is only fluorescent when bound, you can consider to omit washing steps when working with sensitve tissues.

Solutions and Reagents:

EbbaBiolight is provided as 1000-fold concentrated solution. The following common reagents are required (not supplied):

  • Ethanol, 95% (-20°C) or 4% PFA
  • Phosphate buffered saline (PBS), pH 7.4
  • Deionized water
  • Glass coverslips
  • Mounting medium

Assay Procedure:

  • Fix infected cells or tissue sections with method of choice. We recommend fixation with ice-cold ethanol (5 min) at room temperature.
  • Rehydrate tissue sections in a mix of ethanol and deionized water (1:1) for 5 min. The rehydration step may need to be repeated with lower ethanol ratio depending on the tissue.
  • Equilibrate sections in PBS for 5 min.
  • Dilute EbbaBiolight in PBS 1:1000.
  • Apply diluted EbbaBiolight generously. Use enough liquid (ca 0.5 ml) to prevent the sections from drying out during incubation. Incubate for 30 min.
  • Wash 2 x 5 min in PBS (Optional).
  • Mount tissue sections and seal the coverslip onto the slide to prevent drying.

Fluorescence Microscopy:

  • EbbaBiolight 680 is excited at 561 nm (standard laser line) and emission is detected using a standard PI (Propodium Iodide), mCherry or Cy3.5 filter set. Optional: An excitation range of 530-565 nm and a detection range of 600-800 nm may be applied depending on available laser lines and filter sets.

Download Protocol

Protocol III: Biofilm tracking in live cultures using EbbaBiolight 680

This protocol describes how to trace the formation of biofilm in bacterial cultures using EbbaBiolight 680 which binds to the biofilm markers curli and cellulose. As EbbaBiolight 680 does not influence biofilm formation when used in recommended concentrations, it can be present in growing cultures. We have tested EbbaBiolight 680 for tracing biofilm produced by Salmonella Enteritidis and Salmonella Typhimurium during growth. For these strains we have not observed staining of intracellular or membrane components. Please make sure that the pH remains constant during your growth experiment. EbbaBiolight 680 works best at pH 7.4

Solutions and Reagents:

EbbaBiolight 680 is provided as 1000-fold concentrated solution. The following common reagents are required (not supplied):

  • Growth medium
  • Phosphate buffered saline (PBS), pH 7.4
  • 96-well plate (round bottom) with cover
  • Deionized water
  • Spectrophotometer

Assay Procedure:

  • Dilute EbbaBiolight 680 in growth medium 1:1000
  • Inoculate supplemented growth medium with bacterial culture.
  • Fill the wells of the 96-well plate with 100 μl inoculated medium.
  • Fill unused wells with sterile water to avoid drying during incubation.
  • Seal the plate with a cover or adhesive seal.
  • To maximize the temporal resolution when monitoring the biofilm formation, we recommend to incubate the 96-well plate directly in the spectrophotometer. Alternatively, position the 96-well plate in a standard incubator and move it to the spectrophotometer at regular time intervals for recording.

Spectrophotometer Settings:

  • EbbaBiolight 680: Excite at 540 nm and collect emission at 680 nm. Optional: Record an emission spectrum (560 - 800 nm) with 540 nm excitation.

Download Protocol

Protocol IV: Biofilm quantification in colony re-suspensions using EbbaBiolight 680

This protocol describes how to quantify the biofilm markers curli and cellulose in colony re-suspensions using EbbaBiolight 680. We have tested this procedure with Salmonella Enteritidis and Salmonella Typhimurium strains. For these strains we have not observed staining of intracellular or membrane components.

Solutions and Reagents:

EbbaBiolight 680 is provided as 1000-fold concentrated solution. The following common reagents are required (not supplied):

  • Agar plates
  • Phosphate buffered saline (PBS), pH 7.4
  • 96-well plate (round bottom)
  • Spectrophotometer

Assay Procedure:

  • Grow bacterial colonies on an agar plates under biofilm forming conditions. Notice: no morphotyping is required for this procedure.
  • Dilute EbbaBiolight 680™ in PBS 1:1000
  • Add 100 μl into each well of a 96-well plate.
  • Pick bacterial colonies from the agar plate and resuspend thoroughly into each of the pre-filled wells.
  • Place the plate in a spectrophotometer to quantify biofilm in the colony resuspensions.

Spectrophotometer Settings:

  • EbbaBiolight 680: Excite at 540 nm and collect emission at 680 nm. Optional: Record an emission spectrum (560 - 800 nm) with 540 nm excitation.

Download Protocol

Download
Download
Download
Download
Download
Download
Download
Download
Download
Download
Terms & Conditions

For detailed information on Terms & Conditions, please click the link below.

Terms & Conditions

Privacy Policy

To read more how we handle personal information, please click the link below.

Privacy Policy & Cookie Policy

Support

If you have any questions related to our products or services, please contact us.

info@ebbabiotech.com