Dual channel confocal laser scanning microscopy of lucifer yellow-microinjected human brain cells combined with Texas red immunofluorescence.
A method for visualization of individual human brain cells and their dendritic extensions in combination with immunofluorescence is described. Microinjection of Lucifer Yellow was used to reveal the dendritic morphology of cortical brain cells. Indirect immunofluorescence with Texas Red as label was used to investigate the distribution of 3 different groups of immunogens: enzymes (monoamine oxidase A and B), receptors (beta-adrenoceptor protein), and synaptic vesicle proteins (synapsin I and synaptophysin) in each cortical slice. A dual-channel confocal laser scanning microscope with an argon/krypton laser was used for imaging these double-stained fluorescent specimens. Lucifer Yellow and Texas Red were recorded simultaneously or separately, taking advantage of the different activating lines (488 lambda and 568 lambda) of the laser and using the two filter blocks (K1 and K2) supplied with the instrument (BioRad MRC-600) for recording the emission of either fluorophore. Using this technique we have demonstrated the localization of immunoreactive material in relation to the dendritic morphology of cortical cells.
Brain Research Institute, Russian Academy of Medical Sciences, Moscow.
P V
PV
Belichenko
A
A
Dahlström
D001921:Brain / Q000166:cytology*; D002675:Child, Preschool; D005260:Female; D005455:Fluorescent Antibody Technique; D005456:Fluorescent Dyes; D006801:Humans; D007527:Isoenzymes / Q000378:metabolism; D007546:Isoquinolines; D008845:Microinjections; D018613:Microscopy, Confocal; D008995:Monoamine Oxidase / Q000378:metabolism; D009419:Nerve Tissue Proteins / Q000378:metabolism; D010865:Pilot Projects; D011943:Receptors, Adrenergic, beta / Q000378:metabolism; D013572:Synaptic Vesicles / Q000378:metabolism; D014966:Xanthenes
D005456:Fluorescent Dyes; D007527:Isoenzymes; D007546:Isoquinolines; D009419:Nerve Tissue Proteins; D011943:Receptors, Adrenergic, beta; D014966:Xanthenes; C034657:Texas red; C017475:lucifer yellow; D008995:Monoamine Oxidase