PMID- 26948044
OWN - NLM
STAT- MEDLINE
DCOM- 20161214
LR  - 20191210
IS  - 1872-8359 (Electronic)
IS  - 0167-7012 (Linking)
VI  - 124
DP  - 2016 May
TI  - Phytoplankton IF-FISH: Species-specific labeling of cellular proteins by 
      immunofluorescence (IF) with simultaneous species identification by fluorescence 
      immunohybridization (FISH).
PG  - 21-7
LID - S0167-7012(16)30033-1 [pii]
LID - 10.1016/j.mimet.2016.02.017 [doi]
AB  - Phytoplankton rarely occur as unialgal populations. Therefore, to study 
      species-specific protein expression, indicative of physiological status in 
      natural populations, methods are needed that will both assay for a protein of 
      interest and identify the species expressing it. Here we describe a protocol for 
      IF-FISH, a dual labeling procedure using immunofluorescence (IF) labeling of a 
      protein of interest followed by fluorescence in situ hybridization (FISH) to 
      identify the species expressing that protein. The protocol was developed to 
      monitor expression of the cell cycle marker proliferating cell nuclear antigen 
      (PCNA) in the red tide dinoflagellate, Karenia brevis, using a large subunit 
      (LSU) rRNA probe to identify K. brevis in a mixed population of morphologically 
      similar Karenia species. We present this protocol as proof of concept that 
      IF-FISH can be successfully applied to phytoplankton cells. This method is widely 
      applicable for the analysis of single-cell protein expression of any protein of 
      interest within phytoplankton communities.
CI  - Copyright (c) 2016 Elsevier B.V. All rights reserved.
FAU - Meek, Megan E
AU  - Meek ME
AD  - Graduate Program in Marine Biology, Grice Marine Laboratory, College of 
      Charleston, 205 Fort Johnson Rd., Charleston, SC 29412, United States; NOAA 
      Marine Biotoxins Program, Center for Coastal Environmental Health and 
      Biomolecular Research, 219 Fort Johnson Rd., Charleston, SC 29412, United States. 
      Electronic address: meekme@g.cofc.edu.
FAU - Van Dolah, Frances M
AU  - Van Dolah FM
AD  - Graduate Program in Marine Biology, Grice Marine Laboratory, College of 
      Charleston, 205 Fort Johnson Rd., Charleston, SC 29412, United States; NOAA 
      Marine Biotoxins Program, Center for Coastal Environmental Health and 
      Biomolecular Research, 219 Fort Johnson Rd., Charleston, SC 29412, United States.
LA  - eng
PT  - Evaluation Study
PT  - Journal Article
DEP - 20160304
PL  - Netherlands
TA  - J Microbiol Methods
JT  - Journal of microbiological methods
JID - 8306883
RN  - 0 (Protozoan Proteins)
SB  - IM
MH  - Dinoflagellida/chemistry/classification/genetics/*isolation & purification
MH  - Fluorescent Antibody Technique/*methods
MH  - In Situ Hybridization, Fluorescence/*methods
MH  - Phytoplankton/chemistry/classification/genetics/*isolation & purification
MH  - Protozoan Proteins/chemistry/*genetics/metabolism
MH  - Species Specificity
OTO - NOTNLM
OT  - Dinoflagellate
OT  - Flow cytometry
OT  - Immunofluorescence
OT  - In situ hybridization
OT  - Karenia brevis
OT  - Phytoplankton
EDAT- 2016/03/08 06:00
MHDA- 2016/12/15 06:00
CRDT- 2016/03/08 06:00
PHST- 2015/12/22 00:00 [received]
PHST- 2016/02/28 00:00 [revised]
PHST- 2016/02/29 00:00 [accepted]
PHST- 2016/03/08 06:00 [entrez]
PHST- 2016/03/08 06:00 [pubmed]
PHST- 2016/12/15 06:00 [medline]
AID - S0167-7012(16)30033-1 [pii]
AID - 10.1016/j.mimet.2016.02.017 [doi]
PST - ppublish
SO  - J Microbiol Methods. 2016 May;124:21-7. doi: 10.1016/j.mimet.2016.02.017. Epub 
      2016 Mar 4.