Limits...
Profile analysis of hepatic porcine and murine brain tissue slices obtained with a vibratome.

Mattei G, Cristiani I, Magliaro C, Ahluwalia A - PeerJ (2015)

Bottom Line: In particular, the effect of two sectioning parameters (i.e., step size and sectioning speed) on resultant slice thickness was investigated for fresh porcine liver as well as for paraformaldehyde-fixed (PFA-fixed) and fresh murine brain.Conversely, the thickness of PFA-fixed murine brain slices was found to be dependent on the step size, but not on the sectioning speed.In view of these results, fresh brain tissue was sliced varying the step size only, which was found to have a significant effect on resultant slice thickness.

View Article: PubMed Central - HTML - PubMed

Affiliation: Research Center "E. Piaggio," University of Pisa , Pisa , Italy.

ABSTRACT
This study is aimed at characterizing soft tissue slices using a vibratome. In particular, the effect of two sectioning parameters (i.e., step size and sectioning speed) on resultant slice thickness was investigated for fresh porcine liver as well as for paraformaldehyde-fixed (PFA-fixed) and fresh murine brain. A simple framework for embedding, sectioning and imaging the slices was established to derive their thickness, which was evaluated through a purposely developed graphical user interface. Sectioning speed and step size had little effect on the thickness of fresh liver slices. Conversely, the thickness of PFA-fixed murine brain slices was found to be dependent on the step size, but not on the sectioning speed. In view of these results, fresh brain tissue was sliced varying the step size only, which was found to have a significant effect on resultant slice thickness. Although precision-cut slices (i.e., with regular thickness) were obtained for all the tissues, slice accuracy (defined as the match between the nominal step size chosen and the actual slice thickness obtained) was found to increase with tissue stiffness from fresh liver to PFA-fixed brain. This quantitative investigation can be very helpful for establishing the most suitable slicing setup for a given tissue.

No MeSH data available.


Related in: MedlinePlus

Fresh liver slice thicknesses.(A) Bar plot: different letters indicate significant differences between samples (p < 0.05). Different bar fillings indicate different step sizes. (B) Two-way ANOVA interaction plot: the interaction between the step size and the sectioning speed is not significant. Error bars represent standard deviations.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4419543&req=5

fig-3: Fresh liver slice thicknesses.(A) Bar plot: different letters indicate significant differences between samples (p < 0.05). Different bar fillings indicate different step sizes. (B) Two-way ANOVA interaction plot: the interaction between the step size and the sectioning speed is not significant. Error bars represent standard deviations.

Mentions: We were unable to obtain slices using the 100 µm step size because the blade tended to deform and scrape over the tissue rather than cut it, regardless of the sectioning speed. This is likely due to the very labile and floppy nature of fresh liver (Mattei et al., 2014b). Although the two-way ANOVA analysis showed that both the step size and sectioning speed have a significant effect on the resultant thickness of fresh liver slices, 4 of the 6 step size-sectioning speed combinations investigated (specifically 200-0.1, 200-0.4, 400-0.1 and 400-0.2 µm-mm/s) yielded similar slice thicknesses, with an average value of 540 ± 91 µm (Fig. 3A). Moreover, the interaction between the two factors (i.e., step size and sectioning speed) was not found to be significant: lines in Fig. 3B exhibit the same trend versus the sectioning speed, regardless of the step size.


Profile analysis of hepatic porcine and murine brain tissue slices obtained with a vibratome.

Mattei G, Cristiani I, Magliaro C, Ahluwalia A - PeerJ (2015)

Fresh liver slice thicknesses.(A) Bar plot: different letters indicate significant differences between samples (p < 0.05). Different bar fillings indicate different step sizes. (B) Two-way ANOVA interaction plot: the interaction between the step size and the sectioning speed is not significant. Error bars represent standard deviations.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4419543&req=5

fig-3: Fresh liver slice thicknesses.(A) Bar plot: different letters indicate significant differences between samples (p < 0.05). Different bar fillings indicate different step sizes. (B) Two-way ANOVA interaction plot: the interaction between the step size and the sectioning speed is not significant. Error bars represent standard deviations.
Mentions: We were unable to obtain slices using the 100 µm step size because the blade tended to deform and scrape over the tissue rather than cut it, regardless of the sectioning speed. This is likely due to the very labile and floppy nature of fresh liver (Mattei et al., 2014b). Although the two-way ANOVA analysis showed that both the step size and sectioning speed have a significant effect on the resultant thickness of fresh liver slices, 4 of the 6 step size-sectioning speed combinations investigated (specifically 200-0.1, 200-0.4, 400-0.1 and 400-0.2 µm-mm/s) yielded similar slice thicknesses, with an average value of 540 ± 91 µm (Fig. 3A). Moreover, the interaction between the two factors (i.e., step size and sectioning speed) was not found to be significant: lines in Fig. 3B exhibit the same trend versus the sectioning speed, regardless of the step size.

Bottom Line: In particular, the effect of two sectioning parameters (i.e., step size and sectioning speed) on resultant slice thickness was investigated for fresh porcine liver as well as for paraformaldehyde-fixed (PFA-fixed) and fresh murine brain.Conversely, the thickness of PFA-fixed murine brain slices was found to be dependent on the step size, but not on the sectioning speed.In view of these results, fresh brain tissue was sliced varying the step size only, which was found to have a significant effect on resultant slice thickness.

View Article: PubMed Central - HTML - PubMed

Affiliation: Research Center "E. Piaggio," University of Pisa , Pisa , Italy.

ABSTRACT
This study is aimed at characterizing soft tissue slices using a vibratome. In particular, the effect of two sectioning parameters (i.e., step size and sectioning speed) on resultant slice thickness was investigated for fresh porcine liver as well as for paraformaldehyde-fixed (PFA-fixed) and fresh murine brain. A simple framework for embedding, sectioning and imaging the slices was established to derive their thickness, which was evaluated through a purposely developed graphical user interface. Sectioning speed and step size had little effect on the thickness of fresh liver slices. Conversely, the thickness of PFA-fixed murine brain slices was found to be dependent on the step size, but not on the sectioning speed. In view of these results, fresh brain tissue was sliced varying the step size only, which was found to have a significant effect on resultant slice thickness. Although precision-cut slices (i.e., with regular thickness) were obtained for all the tissues, slice accuracy (defined as the match between the nominal step size chosen and the actual slice thickness obtained) was found to increase with tissue stiffness from fresh liver to PFA-fixed brain. This quantitative investigation can be very helpful for establishing the most suitable slicing setup for a given tissue.

No MeSH data available.


Related in: MedlinePlus