Table 1 Integrated light, laser, electron and x-ray analyses used to identify and spatially map the distribution of calcification within human aortic valve leaflets.

From: Osteopontin stabilization and collagen containment slows amorphous calcium phosphate transformation during human aortic valve leaflet calcification

System components

Analyses (resolution)

Histology cryosection

Petrographic section

CT (600 Î¼m) micro-CT (3 Î¼m)

ESEM (5nm)

EDAX (1 Î¼m) ca/p atomic % + / − SD n = 9

Raman (1 Î¼m) spectra cm−1

BF (240 nm) TPMT (240 nm)

BF (240 nm) ORO & ALZ

CPOL (240 nm)

CPOL (240 nm) ALZ

SRAF (140 nm) DAPI

SRAF (140 nm) OPN

Aortic value leaflet tissuse

✓

✓

✓

✓

✓

✓

✓

✓

✓

✓

✓

✓

Lipids

✓

na

na

na

Na

na

na

na

na

na

Red

na

Nuclei

✓

na

na

na

Na

na

na

na

na

na

Blue

na

Osteopontin OPN

✓

na

na

na

Na

na

na

na

na

na

na

Green

Cholesterol crystals

✓

✓

✓

✓

✓

na

✓

Red

1st–2nd order*

1st–2nd order*

Black

✓

Original collagen

✓

✓

✓

✓

Bd

na

✓

na

2nd order*

2nd order*

Blue–green

✓

ACP replaced collagen

✓

✓

✓

✓

1.17–2.49 + / − 0.38

943–958

✓

Red

2nd order*

2nd order*

Red–yellow

✓

ACP nodule

✓

✓

✓

✓

1.47–1.91 + / − 0.15

943–958

✓

Red

2nd order*

2nd order*

Gray–green

✓

  1. Summary of analyses of each of the primary calcification components and analyses. Summary of analyses used to characterize the primary components of aortic valve leaflet calcification.
  2. ACP amorphous calcium phosphate, OPN osteopontin, CT computed tomography, Micro-CT micro-computed tomography, ESEM environmental scanning electron microscopy, EDAX energy dispersive spectroscopy, Raman Raman spectroscopy, BF brightfield microscopy, TPMT transmission photomultiplier tube, ORO Oil Red O red stain for lipids, ALZ Alizarin Red S stain for calcium, DAPI 4′,6-diamidino-2-phenylindole stain for DNA, OPN Alexa 647 antibody green stain for osteopontin, CPOL circular polarization, CPOL ALZ birefringence induced by ALZ stain for calcium, SRAF super resolution autofluorescence, SRIF super resolution induced fluorescence.
  3. ✓ bold = analysis completed, ✓ unbold = delected, na = not applicable, bd = below detected, * = Zesis 2015 Michel-Levy interference colour chart.