PMID- 1581499
OWN - NLM
STAT- MEDLINE
DCOM- 19920618
LR  - 20181113
IS  - 0006-3495 (Print)
IS  - 1542-0086 (Electronic)
IS  - 0006-3495 (Linking)
VI  - 61
IP  - 4
DP  - 1992 Apr
TI  - Calibration of a resonance energy transfer imaging system.
PG  - 845-57
AB  - A quantitative technique for the nondestructive visualization of nanometer scale 
      intermolecular separations in a living system is described. A calibration 
      procedure for the acquisition and analysis of resonance energy transfer (RET) 
      image data is outlined. The factors limiting RET imaging of biological samples 
      are discussed. Measurements required for the calibration include: (a) the 
      spectral sensitivity of the image intensifier (or camera); (b) the transmission 
      spectra of the emission filters; and (c) the quantum distribution functions of 
      the energy transfer pair measured in situ. Resonance energy transfer imaging is 
      demonstrated for two DNA specific dyes. The Forster critical distance for energy 
      transfer between Hoechst 33342 (HO) and acridine orange (AO) is 4.5 +/- 0.7 nm. 
      This distance is slightly greater than the distance of a single turn of the DNA 
      helix (3.5 nm or approximately 10 base pairs), and is well below the optical 
      diffraction limit. Timed sequences of intracellular energy transfer reveal 
      nuclear structure, strikingly similar to that observed with confocal and electron 
      microscopy, and may show the spatial distribution of eu- and hetero- chromatin in 
      the interphase nuclei.
FAU - Ludwig, M
AU  - Ludwig M
AD  - C. W. Bill Young Marrow Donor Recruitment and Research Program, Naval Medical 
      Research Institute, Bethesda, Maryland 20889-5055.
FAU - Hensel, N F
AU  - Hensel NF
FAU - Hartzman, R J
AU  - Hartzman RJ
LA  - eng
PT  - Journal Article
PT  - Research Support, U.S. Gov't, Non-P.H.S.
PL  - United States
TA  - Biophys J
JT  - Biophysical journal
JID - 0370626
RN  - 0 (Benzimidazoles)
RN  - 0 (Fluorescent Dyes)
RN  - 9007-49-2 (DNA)
RN  - F30N4O6XVV (Acridine Orange)
RN  - P976261J69 (bisbenzimide ethoxide trihydrochloride)
SB  - IM
MH  - Acridine Orange
MH  - Benzimidazoles
MH  - Biophysical Phenomena
MH  - Biophysics
MH  - DNA/metabolism
MH  - *Energy Transfer
MH  - *Fluorescent Dyes
MH  - Humans
MH  - Lymphocytes/metabolism
MH  - Spectrometry, Fluorescence
PMC - PMC1260344
EDAT- 1992/04/01 00:00
MHDA- 1992/04/01 00:01
PMCR- 1993/04/01
CRDT- 1992/04/01 00:00
PHST- 1992/04/01 00:00 [pubmed]
PHST- 1992/04/01 00:01 [medline]
PHST- 1992/04/01 00:00 [entrez]
PHST- 1993/04/01 00:00 [pmc-release]
AID - S0006-3495(92)81892-1 [pii]
AID - 10.1016/S0006-3495(92)81892-1 [doi]
PST - ppublish
SO  - Biophys J. 1992 Apr;61(4):845-57. doi: 10.1016/S0006-3495(92)81892-1.