Optimized Fixation and Storage Conditions for FISH Analysis of Single-Cell Suspensions
Author(s): Murrell-Bussell S, Nguyen D, Schober WD, Scott J, Simpson JL, Elias S, Bischoff FZ, Lewis DE
Publication: J Histochem Cytochem, 1998, Vol. 46, Page 971
PubMed ID: 9671447 PubMed Review Paper? No
Purpose of Paper
Conclusion of Paper
Studies
-
Study Purpose
To determine if the distribution of the cell population was affected by fixative concentration (1 vs 3% PF) and subsequent storage in PBS at 4 degrees C (0-21 days).
Summary of Findings:
Forward scatter progressively decreased with PBS storage beginning on day 3 in 1% PF fixed cells and day 7 in all other fixed cells. The reduction in light scatter was less robust in cells fixed with 3% PF, indicating greater stability.
Biospecimens
Preservative Types
- Other Preservative
Diagnoses:
- Not specified
Platform:
Analyte Technology Platform Cell count/volume Flow cytometry Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) Storage Storage duration 0 days
3 days
7 days
10 days
21 days
Biospecimen Preservation Concentration of fixative 1% paraformaldehyde
3% paraformaldehyde
-
Study Purpose
To determine if cell mophology was impacted by fixative concentration and subsequent storage in PBS at 4 degrees C.
Summary of Findings:
The structural integrity of cells fixed in 1% and 3% PF was compromised on day 3 and 21, respectively.
Biospecimens
Preservative Types
- Other Preservative
Diagnoses:
- Not specified
Platform:
Analyte Technology Platform Cell count/volume Flow cytometry Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) Storage Storage duration 0 days
3 days
7 days
10 days
21 days
Biospecimen Preservation Concentration of fixative 1% paraformaldehyde
3% paraformaldehyde
-
Study Purpose
To determine if FISH signal intensity was affected by fixative concentration (1% vs 3% PF) and subsequent storage in PBS at 4 degrees C (0-21 days).
Summary of Findings:
FISH signal intensity, determined by signal/noise ratio, was greatest on day 3 and 10 in cells fixed with 1% and 3% PF for 60 min, respectively. FISH signal clarity in 3% PF fixed cells was superior to those fixed in 1% PF, although signals were stable at both concentrations until day 10, with degradation onset by day 21.
Biospecimens
Preservative Types
- Other Preservative
Diagnoses:
- Not specified
Platform:
Analyte Technology Platform DNA FISH Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) Storage Storage duration 0 days
3 days
7 days
10 days
21 days
Biospecimen Preservation Concentration of fixative 1% paraformaldehyde
3% paraformaldehyde