Targeted imaging modality selection for bacterial biofilms in chronic rhinosinusitis.
OBJECTIVE
Biofilms are increasingly recognized as having an etiological role in chronic rhinosinusitis (CRS). Research into biofilms in CRS currently relies on microscopic imaging techniques, none of which are universally accepted. This study compares LIVE/DEAD BacLight (Invitrogen Corp., Carlsbad, CA) staining and fluorescence in situ hybridization (FISH), both utilizing the confocal scanning laser microscope (CSLM) for biofilm determination and characterization in CRS patients.


METHODS
Prospective study.


METHODS
Twenty CRS patients undergoing endoscopic sinus surgery were recruited for the study. Sinus mucosal tissue harvested at the time of surgery underwent both the BacLight/CSLM and FISH/CSLM protocols for biofilm determination and characterization.


RESULTS
Combining the results of both protocols, 18/20 (90%) patients had bacterial biofilms demonstrable on at least one modality. The high biofilm detection rate combining the two techniques suggests the prevalence of biofilms in CRS may be greater than previously reported. The protocols had equivalent results in 15/20 patients. Using the differences observed in the remaining five patients, we can highlight the most appropriate use for each technique.


CONCLUSIONS
BacLight/CSLM and FISH/CSLM are complementary techniques for biofilm determination and characterization. Both protocols are suited to different research areas and the selection of technique used should be based on the specific objectives of the research protocol. In this way we can utilize the advantages of each technique to facilitate effective research.
Foreman
Andrew
A
Department of Surgery-Otorhinolaryngology, Head and Neck Surgery, University of Adelaide and Flinders University, Adelaide, Australia.
Singhal
Deepti
D
Psaltis
Alkis J
AJ
Wormald
Peter-John
PJ
D001419:Bacteria / Q000302:isolation & purification*; D018441:Biofilms; D002908:Chronic Disease; D005260:Female; D005456:Fluorescent Dyes; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D008297:Male; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D008875:Middle Aged; D009297:Nasal Mucosa / Q000382:microbiology*; D010256:Paranasal Sinuses / Q000382:microbiology* / Q000601:surgery; D012220:Rhinitis / Q000382:microbiology*; D012852:Sinusitis / Q000382:microbiology*
D005456:Fluorescent Dyes