PMID- 10192042
OWN - NLM
STAT- MEDLINE
DCOM- 19990706
LR  - 20191103
IS  - 0167-7012 (Print)
IS  - 0167-7012 (Linking)
VI  - 35
IP  - 2
DP  - 1999 Mar
TI  - Quantitative imaging and statistical analysis of fluorescence in situ 
      hybridization (FISH) of Aureobasidium pullulans.
PG  - 101-10
AB  - Image and multifactorial statistical analyses were used to evaluate the intensity 
      of fluorescence signal from cells of three strains of A. pullulans and one strain 
      of Rhodosporidium toruloides, as an outgroup, hybridized with either a universal 
      or an A. pullulans 18S rRNA oligonucleotide probe in direct or indirect FISH 
      reactions. In general, type of fixation (paraformaldehyde or methanol-acetic 
      acid) had no apparent effect on cell integrity and minimal impact on 
      fluorescence. Permeabilization by enzyme treatment for various times, though 
      needed to admit high Mw detection reagents (avidin-FITC) in indirect FISH, tended 
      to nonspecifically degrade cells and lower the signal. Digestion was unnecessary 
      and undesirable for the directly labelled probes. Multilabelled (five fluorescein 
      molecules) probes enhanced fluorescence about fourfold over unilabelled probes. 
      Overall, direct FISH was preferable to indirect FISH and is recommended 
      especially for studies of microbes on natural substrata.
FAU - Spear, R N
AU  - Spear RN
AD  - Department of Plant Pathology, University of Wisconsin Madison 53706, USA.
FAU - Li, S
AU  - Li S
FAU - Nordheim, E V
AU  - Nordheim EV
FAU - Andrews, J H
AU  - Andrews JH
LA  - eng
PT  - Journal Article
PT  - Research Support, U.S. Gov't, Non-P.H.S.
PL  - Netherlands
TA  - J Microbiol Methods
JT  - Journal of microbiological methods
JID - 8306883
RN  - 0 (Oligonucleotide Probes)
RN  - 0 (RNA, Ribosomal, 18S)
SB  - IM
MH  - Analysis of Variance
MH  - Cell Wall/metabolism
MH  - *Image Processing, Computer-Assisted
MH  - In Situ Hybridization, Fluorescence/*methods
MH  - Mitosporic Fungi/*genetics
MH  - Oligonucleotide Probes
MH  - RNA, Ribosomal, 18S/genetics
EDAT- 1999/04/07 00:00
MHDA- 1999/04/07 00:01
CRDT- 1999/04/07 00:00
PHST- 1999/04/07 00:00 [pubmed]
PHST- 1999/04/07 00:01 [medline]
PHST- 1999/04/07 00:00 [entrez]
AID - S0167701298001006 [pii]
AID - 10.1016/s0167-7012(98)00100-6 [doi]
PST - ppublish
SO  - J Microbiol Methods. 1999 Mar;35(2):101-10. doi: 10.1016/s0167-7012(98)00100-6.