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    Histology and Cytology

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    Dharshini Kumar
    The document describes the lipid stain technique used to identify neutral lipids and fatty acids in tissue sections. The Oil Red O stain is used, which demonstrates lipids as red and nuclei as blue. Fresh or frozen sections are required, as fixatives containing alcohols will remove lipids. The procedure involves staining sections with Oil Red O, counterstaining with hematoxylin, and mounting in glycerin jelly. Lipoproteins may be demonstrated on paraffin sections.

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    Histology and Cytology

    Uploaded by

    Dharshini Kumar
    27 views4 pages
    The document describes the lipid stain technique used to identify neutral lipids and fatty acids in tissue sections. The Oil Red O stain is used, which demonstrates lipids as red and nuclei as blue. Fresh or frozen sections are required, as fixatives containing alcohols will remove lipids. The procedure involves staining sections with Oil Red O, counterstaining with hematoxylin, and mounting in glycerin jelly. Lipoproteins may be demonstrated on paraffin sections.

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    Good notes from national institute of open schooling

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    Histology and cytology

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    The document describes the lipid stain technique used to identify neutral lipids and fatty acids in tissue sections. The Oil Red O stain is used, which demonstrates lipids as red and nuclei as blue. Fresh or frozen sections are required, as fixatives containing alcohols will remove lipids. The procedure involves staining sections with Oil Red O, counterstaining with hematoxylin, and mounting in glycerin jelly. Lipoproteins may be demonstrated on paraffin sections.

    Copyright:

    © All Rights Reserved
    Download as PDF, TXT or read online from Scribd
    Download as pdf or txt
    27 views4 pages

    Histology and Cytology

    Uploaded by

    Dharshini Kumar
    The document describes the lipid stain technique used to identify neutral lipids and fatty acids in tissue sections. The Oil Red O stain is used, which demonstrates lipids as red and nuclei as blue. Fresh or frozen sections are required, as fixatives containing alcohols will remove lipids. The procedure involves staining sections with Oil Red O, counterstaining with hematoxylin, and mounting in glycerin jelly. Lipoproteins may be demonstrated on paraffin sections.

    Copyright:

    © All Rights Reserved
    Download as PDF, TXT or read online from Scribd
    Download as pdf or txt
    of 4

    Lipid Stain MODULE

    Histology and Cytology

    13
    Notes
    LIPID STAIN

    13.1 INTRODUCTION
    The Oil Red O (ORO) stain can identify neutral lipids and fatty acids in smears
    and tissues. Fresh smears or cryostat sections of tissue are necessary because
    fixatives containing alcohols, or routine tissue processing with clearing, will
    remove lipids. The ORO is a rapid and simple stain.

    OBJECTIVES
    After reading this lesson, you will be able to:
    z explain the principle of lipid stain
    z describe various reagents used for lipid stains
    z describe the procedure of lipid staining.

    13.2 LIPID STAIN


    Aim: To demonstrate intracellular lipid in tissue sections.
    Principle: The dye being more soluble in the lipid to be demonstrated than in
    the vehicular solvent. The polyazo group of dyes includes the oil red series, the
    sudan red series and sudan blacks. All these dyes are interchangeable and may
    be substituted.
    Sudan series - Sudan III
    - Sudan IV
    - Sudan black
    Control - Lipid positive section

    HISTOLOGY AND CYTOLOGY 83


    MODULE Lipid Stain

    Histology and Cytology


    Reagents
    1. Oil Red O stock solution -
    Oil Red O 0.5gm
    Isopropanol 100ml
    Dissolve the dye in isopropanol using gentle heat in water bath.
    Notes 2. Oil Red O working solution
    Stock Oil Red O solution 30ml
    Distilled water 20ml
    Dilute the stock solution with distilled water and keep it for 10 minutes,
    filter and cover it immediately.
    3. Glycerine Jelly Mounting medium
    Gelatin 10gm
    Distilled water 60ml
    Glycerol 70ml
    Phenol 0.25gm
    Dissolve the gelatin in distilled water using sufficient heat to melt the
    gelatin, add glycerol and phenol. Mix well and transfer to a small capped
    bottle and refrigerate.

    Procedure
    z Fix timer in formalin, wash with running tap water for 5 to 10 minutes.
    z Cut frozen section of 8 to 10 micron thickness and air dry.
    z Rinse with 60% isopropanol.
    z Stain with freshly prepared Oil Red O working solution for 15 minutes.
    z Rinse with 60% isopropanol.
    z Few dips in Alum hematoxylin to stain nuclei.
    z Rinse with distilled water.
    z Mount in water or glycerine jelly.

    Result
    z Lipid red
    z Nuclei blue

    84 HISTOLOGY AND CYTOLOGY


    Lipid Stain MODULE
    Note Histology and Cytology

    z Use cryostat sections of 8 to 10 micron thickness or formalin fixed smears.


    z working Oil Red O solution should be freshly prepared from stock solution
    and kept in close container.
    z Never take the sections through clearing solvent prior to mounting as this
    will remove the lipid to be demonstrated. Notes
    z Frozen sections should be used to stain neutral triglycerides.
    z Lipoproteins may be demonstrated on paraffin sections.
    z Alcohol fixation removes most lipids.

    INTEXT QUESTIONS 13.1


    1. Dyes used in lipid stain is .....................
    2. ..................... is used to stain nuclei
    3. Lipid is demonstrated by ..................... colour
    4. Nuclei is demonstrated by ..................... colour
    5. Lipoproteins may be demonstrated on ..................... section
    6. ..................... fixation removes lipids

    WHAT HAVE YOU LEARNT


    z Lipid stain is used to demonstrate intracellular lipid in tissue sections
    z Polyazo group of dyes like oil red series, sudan red series and sudan blacks
    are the dye used for demonstrating lipids in tissue section
    z The frozen section should be cut 8 to 10 micron thickness
    z Lipids appear as red and nuclei appear blue
    z Lipoproteins may be demonstrated on paraffin sections
    z Alcohol fixation removes most lipids

    HISTOLOGY AND CYTOLOGY 85


    MODULE Lipid Stain

    Histology and Cytology

    TERMINAL QUESTIONS
    1. What is principle of lipid stain?
    2. Name three dyes used to demonstrate lipid in tissue sections.
    3. What precautions should be observed during lipid staining on tissue
    Notes sections?
    4. What should be the thickness of sections for lipid staining?
    5. What is the mounting media used in lipid staining?

    ANSWERS TO INTEXT QUESTIONS

    13.1
    1. Sudan series
    2. Alum hematoxylin
    3. Red
    4. Blue
    5. Paraffin
    6. Alcohol

    86 HISTOLOGY AND CYTOLOGY

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