FLUORESCENCE MICROSCOPY
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FLUORESCENCE MICROSCOPY. IMMUNOFLUORESCENCE. Multiple immunofluorescence labelling of formalin-fixed paraffin-embedded (FFPE) tissue David Robertson, Kay Savage, Jorge S Reis-Filho and Clare M Isacke*

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FLUORESCENCE MICROSCOPY

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Fluorescence microscopy

FLUORESCENCE MICROSCOPY


Fluorescence microscopy

IMMUNOFLUORESCENCE


Fluorescence microscopy

Multiple immunofluorescence labelling of formalin-fixed

paraffin-embedded (FFPE) tissue

David Robertson, Kay Savage, Jorge S Reis-Filho and Clare M Isacke*

Address: Breakthrough Breast Cancer Research Centre, The Institute of Cancer Research, 237 Fulham Road, London SW3 6JB, UK

PUBLISHED 2008


Fluorescence microscopy

DUCTULES

BREAST OF A PREGNANT MAMMAL


Fluorescence microscopy

MILK PRODUCTION

LOBULE

DUCTULE

DUCT

MILK SECRETION


Fluorescence microscopy

DUCTULE


Fluorescence microscopy

NUCLEUS OF SECRETORY CELL


Fluorescence microscopy

NUCLEUS OF MYOEPITHELIAL CELL


Fluorescence microscopy

SEM OF BREAST ALVEOLUS (OR ACINUS)

SECRETORY CELL

MYOEPITHIAL CELL

FROM OUTSIDE ALVEOLUS

FROM INSIDE ALVEOLUS


Fluorescence microscopy

BREAST SECRETORY CELL


Fluorescence microscopy

LIPID DROPLETS BEING SECRETED FROM THE SECRETORY CELLS OF BREAST DUCTULE EPITHELIUM

LIPID DROPLET BEING EXTRUDED (IT IS COVERED IN PLASMA MEMBRANE)

SECRETORY CELL

MICROVILLI


Fluorescence microscopy

THE PLASMA MEMBRANE AROUND THE SECRETED FAT DROPLETS PREVENTS THEM FROM FLORMING CLUMPS IN THE MILK

BUTTER

BUTTER CHURN


Fluorescence microscopy

FOR COMBINED IMAGE OF MULTIPLE FLUORESCENT ANTIBODIES (DIIFERENT COLOURED FLUOROPHORES):

PHOTOGRAPHS TAKEN WITH DIFFERENT EXCITATION LIGHTS MUST BE TAKEN AND THEN OVERLAPPED (“MERGED”)


Fluorescence microscopy

MYOEPITHELIAL CELL

LUMEN OF DUCTULE

NUCLEUS OF SECRETORY CELL

INTERMEDIATE FILAMENTS OF SECRETORY CELLS

ANTI- SMOOTH MUSCLE ACTIN

ANTI-GLANDULAR CELL KERATIN

DAPI (DNA-STAINING)


Fluorescence microscopy

DETECTION OF BREAST CANCER BY IMMUNOFLUORESCENCE

THIS PARTICULAR CANCER IS DUE TO UNCONTROLLED GROWTH OF

THE EPITHELIAL CELLS OF THE MILK DUCTS

ANTIBODY AGAINSTESTROGEN RECEPTOR

ANTIBODY AGAINSTSECRETORY CELL KERATINS

CANCEROUS BUT NOT YET INVASIVE

NON-CANCEROUS

INVASIVE CANCER


Fluorescence microscopy

PEROXISOMES IN LIVER CELLS


Fluorescence microscopy

HeLA CELLS

DAPI notice light and darker staining areas in the nucleus

(HETEROCHROMATIN / EUCHROMATIN)

Anti-ACTIN ANTIBODY


Fluorescence microscopy

BULL SPERM

DAPI


Fluorescence microscopy

HeLA CELLS

NUCLEUS BLUE; GOLGI BODIES GREEN;MITOCHONDRIA RED


Fluorescence microscopy

RAT URINARY EPITHELIAL CELLS

Anti- CYTOCHROME OXIDASE ANTIBODY

Anti- ACTIN ANTIBODY

DAPI


Fluorescence microscopy

HeLa CELLS

DAPI

Anti - ACTIN

Anti - TUBILIN


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