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


IMMUNOFLUORESCENCE


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


DUCTULES

BREAST OF A PREGNANT MAMMAL


MILK PRODUCTION

LOBULE

DUCTULE

DUCT

MILK SECRETION


DUCTULE


NUCLEUS OF SECRETORY CELL


NUCLEUS OF MYOEPITHELIAL CELL


SEM OF BREAST ALVEOLUS (OR ACINUS)

SECRETORY CELL

MYOEPITHIAL CELL

FROM OUTSIDE ALVEOLUS

FROM INSIDE ALVEOLUS


BREAST SECRETORY CELL


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


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

BUTTER

BUTTER CHURN


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

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


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)


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


PEROXISOMES IN LIVER CELLS


HeLA CELLS

DAPI notice light and darker staining areas in the nucleus

(HETEROCHROMATIN / EUCHROMATIN)

Anti-ACTIN ANTIBODY


BULL SPERM

DAPI


HeLA CELLS

NUCLEUS BLUE; GOLGI BODIES GREEN;MITOCHONDRIA RED


RAT URINARY EPITHELIAL CELLS

Anti- CYTOCHROME OXIDASE ANTIBODY

Anti- ACTIN ANTIBODY

DAPI


HeLa CELLS

DAPI

Anti - ACTIN

Anti - TUBILIN


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