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Biofilm Tissue-Clearing Reagents

Biofilm-Clearing Reagent: iCBiofilm

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Advantages

  • Can make biofilm transparent by simply adding iCBiofilm.
  • Suitable for various kinds of biofilm.
  • Cleared biofilm can be observed by light-sheet fluorescent microscopy (LSFM) or confocal laser-scanning microscopy (CLSM).

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Example of observation after useing iCBiofilm

Optical observation images of MRSA after the addition of PBS or iCBiofilm-H1

Figure. Images of methicillin-resistant Staphylococcus aureus (MRSA) observed microscopically after incubated overnight at 37°C,
fixed at room temperature for 30 minutes, stained with FM®1-43 at 4°C overnight, washed, and dipped in PBS or iCBiofilm-H1.
(FM is a trademark of Thermo Fisher Scientific Inc.)

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Application

iCBioflim-H1 (Product No. 4031) is applicable to clearing many kinds of biofilms. If the transparency with iCBiofilm-H1 is insufficient, iCBiofilm-H2 (Product No. T4032) can be used instead. Since iCBiofilm-H2 is prepared at high component concentration, iCBiofilm-H2 is available as a stock solution. Before addition to biofilm, iCBiofilm-H2 has to be diluted with deionized water to the appropriate concentration. (Undiluted iCBiofilm-H2 may work well.)

[Direction for Use]

  1. Prepare a biofilm in a glass bottomed culture dish (35mm diam.).
  2. After removing planktonic cells and the culture medium from the dish, add 1% glutaraldehyde solution diluted with PBS to fix the biofilm at room temperature for 30 minutes. To preserve immunogenicity for immunofluorescence applications, the use of 4% paraformaldehyde in PBS is recommended.
  3. Remove the glutaraldehyde solution and wash three times with PBS or distilled water.
  4. Stain the biofilm with appropriate fluorescence reagents.
  5. Remove staining reagents and add iCBiofilm solution.
  6. Observe the biofilm by optical microscopy.

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Video showing how to use

This products are commercialized under instruction of Assoc. Prof. Shinya Sugimoto.
Figures and movie were provided by Assoc. Prof. Shinya Sugimoto.

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Product Brochure

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產品號碼:   T4032 | 純度/分析方法  

產品號碼:   T4031 | 純度/分析方法