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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)
MYOEPITHIAL
CELL
SECRETORY
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 CHURN
BUTTER
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 AGAINST ESTROGEN RECEPTOR
ANTIBODY AGAINST SECRETORY CELL KERATINS
NON-CANCEROUS
CANCEROUS BUT
NOT YET INVASIVE
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
Anti - ACTIN
Anti - TUBILIN
DAPI