PMID- 19280050
OWN - NLM
STAT- MEDLINE
DCOM- 20090420
LR  - 20191210
IS  - 0143-2044 (Print)
IS  - 0143-2044 (Linking)
VI  - 29
IP  - 6
DP  - 2008 Nov-Dec
TI  - Evaluation of zebrafish (Danio rerio) ovarian follicle viability by simultaneous 
      staining with fluorescein diacetate and propidium iodide.
PG  - 463-75
AB  - Reliable fish oocyte quality assessment methods are essential in developing 
      protocols for cryopreservation as well as their in vitro maturation and 
      fertilisation. Current ovarian follicle viability assessment methods either lack 
      sensitivity (e.g. Trypan Blue staining-TB) or are stage dependent (e.g. in vitro 
      maturation and observation of germinal vesicle breakdown-GVBD). The aim of the 
      present study was to develop a new viability assessment method for zebrafish 
      ovarian follicles that is reliable, sensitive and not-stage specific. Fluorescein 
      Diacetate (FDA) and Propidium Iodide (PI) were used for the first time to assess 
      viability of zebrafish ovarian follicles. After preliminary studies to evaluate 
      the efficacy of FDA and PI, a combination of these two fluorochromes was 
      subsequently chosen and compared with TB staining and GVBD test in a series of 
      cryoprotectant toxicity studies and following cryopreservation using stage III 
      ovarian follicles. In all cases the FDA-PI test proved to be more sensitive than 
      TB staining but less sensitive than the GVBD test. Ovarian follicle survivals 
      after 4M Methanol treatment for 30 min at 22 degree C were 67.4 +/- 4.4%, 43.9 
      +/- 3.8% and 19.6 +/- 1.9% using TB, FDA-PI and GVBD test respectively. Survivals 
      after cryopreservation procedure were 38.9 +/- 4.0% and 28.9 +/- 2.5% using TB 
      and FDA-PI respectively when Hank's solution was used as medium and 45.2 +/- 4.3% 
      and 35.2 +/- 3.5% when KCl buffer was used. The results showed the method to be 
      promising, and it may offer a new approach for viability assessment of fish 
      ovarian follicles.
FAU - Zampolla, T
AU  - Zampolla T
AD  - LIRANS Institute of Research in the Applied Natural Sciences, University of 
      Bedfordshire, Luton, Bedfordshire, UK.
FAU - Zhang, T
AU  - Zhang T
FAU - Rawson, D M
AU  - Rawson DM
LA  - eng
PT  - Evaluation Study
PT  - Journal Article
PT  - Validation Study
PL  - England
TA  - Cryo Letters
JT  - Cryo letters
JID - 9891832
RN  - 0 (Buffers)
RN  - 0 (Coloring Agents)
RN  - 0 (Fluoresceins)
RN  - 36015-30-2 (Propidium)
RN  - YL39R93PRE (diacetylfluorescein)
SB  - IM
MH  - Animals
MH  - Buffers
MH  - Cell Survival
MH  - Coloring Agents
MH  - *Cryopreservation
MH  - Cytoplasmic Vesicles
MH  - Female
MH  - *Fluoresceins
MH  - Ovarian Follicle/*cytology
MH  - *Propidium
MH  - Reproducibility of Results
MH  - Staining and Labeling/*methods/standards
MH  - Zebrafish
EDAT- 2009/03/13 09:00
MHDA- 2009/04/21 09:00
CRDT- 2009/03/13 09:00
PHST- 2009/03/13 09:00 [entrez]
PHST- 2009/03/13 09:00 [pubmed]
PHST- 2009/04/21 09:00 [medline]
PST - ppublish
SO  - Cryo Letters. 2008 Nov-Dec;29(6):463-75.