PMID- 8580199
OWN - NLM
STAT- MEDLINE
DCOM- 19960321
LR  - 20191101
IS  - 1052-0295 (Print)
IS  - 1052-0295 (Linking)
VI  - 70
IP  - 4
DP  - 1995 Jul
TI  - A color image analysis method for assessment of germination based on differential 
      fluorescence staining of bacterial spores and vegetative cells using acridine 
      orange.
PG  - 175-84
AB  - Color fluorescence image analysis of acridine orange (AO) stained germinating 
      Bacillus subtilis var. niger bacteria revealed a cell population initially 
      dominated by small green spores followed by the emergence of at least three 
      additional discernible subpopulations in response to stimulation with D-glucose. 
      These subpopulations were small, round or oblong red cells; intermediate to large 
      metachromatic cells; and large red rods. Large green rods were rarely observed. 
      An increase in red emissions (i.e., putative RNA synthesis) was sometimes seen as 
      early as 90 min after exposure to D-glucose and uptake of AO at room temperature. 
      This may represent either metabolic recovery from quiescence or RNA synthesis 
      associated with germination. In the absence of D-glucose, or using autoclaved 
      bacteria in the presence of glucose, no relative increase in the red signal was 
      observed despite hours of observation. Digital image analysis was used for 
      relative measurement of red, green and blue signals and to correlate the size of 
      various subpopulations with their fluorescence color emissions over time. Image 
      analysis demonstrated a trend toward increasing size and red emission in the 
      presence of glucose. The average red emission was found to be a good 
      discriminator of the various subpopulations, while the average green emission was 
      approximately equal among the subpopulations making it a poor discriminator. 
      These data suggest that AO staining might be used for rapid computer-assisted 
      discrimination of spores vs. vegetative cells.
FAU - Bruno, J G
AU  - Bruno JG
AD  - U.S. Air Force, Armstrong Laboratory, Edgewood Research, Development and 
      Engineering Center, SCBRD-RTE, Maryland 21010-5423, USA.
FAU - Mayo, M W
AU  - Mayo MW
LA  - eng
PT  - Journal Article
PL  - England
TA  - Biotech Histochem
JT  - Biotechnic & histochemistry : official publication of the Biological Stain 
      Commission
JID - 9107378
RN  - F30N4O6XVV (Acridine Orange)
SB  - IM
MH  - Acridine Orange/*pharmacokinetics
MH  - Bacillus subtilis/metabolism/*physiology
MH  - Image Processing, Computer-Assisted/*methods
MH  - Microscopy, Phase-Contrast/methods
MH  - Spectrometry, Fluorescence/methods
MH  - Spores, Bacterial
EDAT- 1995/07/01 00:00
MHDA- 1995/07/01 00:01
CRDT- 1995/07/01 00:00
PHST- 1995/07/01 00:00 [pubmed]
PHST- 1995/07/01 00:01 [medline]
PHST- 1995/07/01 00:00 [entrez]
AID - 10.3109/10520299509107309 [doi]
PST - ppublish
SO  - Biotech Histochem. 1995 Jul;70(4):175-84. doi: 10.3109/10520299509107309.