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Iodine vapor staining for atomic number contrast in backscattered electron and X-ray imaging.

Boyde A, Mccorkell FA, Taylor GK, Bomphrey RJ, Doube M - Microsc. Res. Tech. (2014)

Bottom Line: Contrast is comparable to that achieved with liquid staining but without the consequent tissue shrinkage, stain pooling, or uneven coverage artefacts associated with immersing the specimen in iodine solutions.Here, we have used iodine vapor staining for two imaging modalities in frequent use in our laboratories and on the specimen types with which we work.It is likely to be equally convenient for a wide range of specimens, and for other modalities which generate contrast from electron- and photon-sample interactions, such as transmission electron microscopy and light microscopy.

View Article: PubMed Central - PubMed

Affiliation: Dental Physical Sciences, Barts and The London School of Medicine and Dentistry, Queen Mary University of London, London, E1 4NS, United Kingdom.

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BSE SEM of iodine vapor stained PMMA-embedded musculoskeletal tissues. Rat distal femur growth plate (distal top). 20 μm scale bar in meta-data panel. Asterix *in marrow space in epiphysis. PMMA block stained with iodine vapor for 2 months.
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fig04: BSE SEM of iodine vapor stained PMMA-embedded musculoskeletal tissues. Rat distal femur growth plate (distal top). 20 μm scale bar in meta-data panel. Asterix *in marrow space in epiphysis. PMMA block stained with iodine vapor for 2 months.

Mentions: Iodine vapor staining results in differential contrast in BSE SEM imaging (Figs. 4).1 Muscle and erythrocytes give strong positive contrast, with chondrocyte nuclei and cartilage matrix giving moderate positive contrast. Pericytic and interterritorial regions of cartilage matrix can be distinguished. Annulus fibrosus and nucleus pulposus of the intervertebral disk stain well. Adipocyte cell peripheries can be clearly distinguished. PMMA itself acquires a small increase in backscattering signal, but not so much as to interfere with reading tissue architecture. Iodine staining is limited to the specimen surface. Cracks do not acquire the high signal seen as an artefact with triiodide solution staining (Boyde, 2012; Ley et al., 2014).


Iodine vapor staining for atomic number contrast in backscattered electron and X-ray imaging.

Boyde A, Mccorkell FA, Taylor GK, Bomphrey RJ, Doube M - Microsc. Res. Tech. (2014)

BSE SEM of iodine vapor stained PMMA-embedded musculoskeletal tissues. Rat distal femur growth plate (distal top). 20 μm scale bar in meta-data panel. Asterix *in marrow space in epiphysis. PMMA block stained with iodine vapor for 2 months.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4285820&req=5

fig04: BSE SEM of iodine vapor stained PMMA-embedded musculoskeletal tissues. Rat distal femur growth plate (distal top). 20 μm scale bar in meta-data panel. Asterix *in marrow space in epiphysis. PMMA block stained with iodine vapor for 2 months.
Mentions: Iodine vapor staining results in differential contrast in BSE SEM imaging (Figs. 4).1 Muscle and erythrocytes give strong positive contrast, with chondrocyte nuclei and cartilage matrix giving moderate positive contrast. Pericytic and interterritorial regions of cartilage matrix can be distinguished. Annulus fibrosus and nucleus pulposus of the intervertebral disk stain well. Adipocyte cell peripheries can be clearly distinguished. PMMA itself acquires a small increase in backscattering signal, but not so much as to interfere with reading tissue architecture. Iodine staining is limited to the specimen surface. Cracks do not acquire the high signal seen as an artefact with triiodide solution staining (Boyde, 2012; Ley et al., 2014).

Bottom Line: Contrast is comparable to that achieved with liquid staining but without the consequent tissue shrinkage, stain pooling, or uneven coverage artefacts associated with immersing the specimen in iodine solutions.Here, we have used iodine vapor staining for two imaging modalities in frequent use in our laboratories and on the specimen types with which we work.It is likely to be equally convenient for a wide range of specimens, and for other modalities which generate contrast from electron- and photon-sample interactions, such as transmission electron microscopy and light microscopy.

View Article: PubMed Central - PubMed

Affiliation: Dental Physical Sciences, Barts and The London School of Medicine and Dentistry, Queen Mary University of London, London, E1 4NS, United Kingdom.

Show MeSH