PMID- 7679591
OWN - NLM
STAT- MEDLINE
DCOM- 19930323
LR  - 20131121
IS  - 1059-910X (Print)
IS  - 1059-910X (Linking)
VI  - 24
IP  - 1
DP  - 1993 Jan 1
TI  - Photoconversion of diaminobenzidine with different fluorescent neuronal markers 
      into a light and electron microscopic dense reaction product.
PG  - 2-14
AB  - This article describes methods for photoconverting diaminobenzidine (DAB) into a 
      stable, light and electron microscopically visible dark reaction product in 
      neurons which contain a fluorescent dye. Photoconversion of DAB has been achieved 
      so far with the following fluorescent dyes: rhodamine labeled latex microspheres 
      (RLM), 4,6-diamidino-2-phenylindole (DAPI), 5,7-dihydroxytryptamine (5,7-DHT), 
      Fast Blue (FB), Nuclear Yellow (NY), Diamidino Yellow (DY), Evans Blue (EB), 
      acridine orange (AO), ethidium bromide (EBR),1,1'-dioctadecyl- 
      3,3,3',3'-tetramethylindolcarbocyanine perchlorate, D-282 (DiI), propidium iodide 
      (PI), and intracellularly injected Lucifer Yellow (LY). The dye is introduced 
      into the neurons by tinctorial staining, retrograde transport, or intracellular 
      injection. Photoconversion is conducted by incubating the tissue with the 
      fluorescent substance-containing cells in a DAB solution under simultaneous 
      strong illumination with ultraviolet (UV) light. During the formation of the 
      reaction product, the fluorescence disappears from the cell. In all cases, 
      photoconversion provided a stable, nonfading DAB reaction product for light 
      microscopy. In addition, at the electron microscopic level, it appeared that the 
      photoconversion results in a homogeneously distributed, fine granular, dark, 
      intracellularly located reaction product. With most of the retrograde tracers 
      tested, photoconversion led only to staining of the cell bodies and the proximal 
      portions of primary dendrites. Following photoconversion with intracellularly 
      LY-filled neurons and cells labeled retrogradely with DiI, DiO, and 5,7-DHT, the 
      reaction product was present throughout the cells, extending from the cell bodies 
      into dendrites and dendritic appendices, and into axons. The high selectivity and 
      methodological simplicity of photoconversion of DAB with fluorescent dyes into a 
      stable, light and electron microscopical dense reaction product provide a 
      promising alternative to classical neuroanatomical techniques and a new useful 
      application of fluorescent neuronal tracers to light and electron microscopy.
FAU - Lubke, J
AU  - Lubke J
AD  - Department of Human Anatomy, University of Oxford, United Kingdom.
LA  - eng
PT  - Journal Article
PL  - United States
TA  - Microsc Res Tech
JT  - Microscopy research and technique
JID - 9203012
RN  - 0 (Fluorescent Dyes)
RN  - 2RV4T6KHQI (3,3'-Diaminobenzidine)
SB  - IM
MH  - 3,3'-Diaminobenzidine/*radiation effects
MH  - Animals
MH  - Brain/ultrastructure
MH  - Cats
MH  - Chick Embryo
MH  - Fluorescent Dyes/*radiation effects
MH  - Light
MH  - Neurons/*ultrastructure
MH  - Staining and Labeling/methods
MH  - Ultraviolet Rays
EDAT- 1993/01/01 00:00
MHDA- 1993/01/01 00:01
CRDT- 1993/01/01 00:00
PHST- 1993/01/01 00:00 [pubmed]
PHST- 1993/01/01 00:01 [medline]
PHST- 1993/01/01 00:00 [entrez]
AID - 10.1002/jemt.1070240103 [doi]
PST - ppublish
SO  - Microsc Res Tech. 1993 Jan 1;24(1):2-14. doi: 10.1002/jemt.1070240103.