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  • Total Lipids Assays Nile Red Staining

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    Фахрия Мамедова
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    This document summarizes a method for determining lipid content in algae using Nile Red staining. It describes preparing a Nile Red solution by dissolving 250 mg of Nile Red in 40 mL of acetone. 10 mL of algal suspension is then mixed with the Nile Red solution and agitated. After 10 minutes, fluorescence intensity is measured using a fluorimeter with 490 nm excitation and 585 nm emission filters. The fluorescence intensity from Nile Red is obtained by subtracting autofluorescence of algal cells and fluorescence from Nile Red alone in the medium.

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    This document summarizes a method for determining lipid content in algae using Nile Red staining. It describes preparing a Nile Red solution by dissolving 250 mg of Nile Red in 40 mL of acetone. 10 mL of algal suspension is then mixed with the Nile Red solution and agitated. After 10 minutes, fluorescence intensity is measured using a fluorimeter with 490 nm excitation and 585 nm emission filters. The fluorescence intensity from Nile Red is obtained by subtracting autofluorescence of algal cells and fluorescence from Nile Red alone in the medium.

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    14 views1 page

    Total Lipids Assays Nile Red Staining

    Uploaded by

    Фахрия Мамедова
    This document summarizes a method for determining lipid content in algae using Nile Red staining. It describes preparing a Nile Red solution by dissolving 250 mg of Nile Red in 40 mL of acetone. 10 mL of algal suspension is then mixed with the Nile Red solution and agitated. After 10 minutes, fluorescence intensity is measured using a fluorimeter with 490 nm excitation and 585 nm emission filters. The fluorescence intensity from Nile Red is obtained by subtracting autofluorescence of algal cells and fluorescence from Nile Red alone in the medium.

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    Pavlo Bohutskyi bohutskyi@jhu.

    edu
    Department of Geography and Environmental Engineering www.bohutskyi.weebly.com
    Johns Hopkins University

    TOTAL LIPIDS ASSAYS – NILE RED STAINING FOR ALGAL LIPID


    DETERMINATION
    Seog June Lee, Byung-Dae Yoon and Hee-Mock Oh. Rapid method for the determination of
    lipid from the green alga Botryococcus braunii. Biotechnology Techniques, Vol 12, No 7,
    July 1998, pp. 553–556

    “Reagent: Nile Red (9-diethylamino-5H-benzo[a]phenoxazine-5-one) from Sigma.

    Nile Red solution, 40 ml, in acetone (250 mg/L) was added to 10 ml of algal suspension. The
    mixture was vigorously agitated on a vortex mixer. Fluorimetric analysis occurred 10 min after
    staining using a digital fluorimeter with a 490 nm narrow band excitation filter and a 585 nm
    narrow band emission filter. The relative fluorescence intensity of Nile Red was obtained after
    subtraction of both the autofluorescence of algal cells and the fluorescence intensity of Nile Red
    alone in the medium”.

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