Considering the demand for investigating spatial cell arrangement within a scaffold, yet unsatisfied, we aimed at developing a new method of sample preparation for SEM that would, besides other things, allow visualization of cellular ultrastructure and deliver from toxic chemicals and artifacts. A combination of BSE imaging in a low-vacuum mode with supravital lanthanoid staining has yielded highly informative images that contain many of the subsurface cell structures, particularly, intercellular contacts, cell membranes, nuclei with nucleoli, mitochondria, endoplasmic reticulum, and cytoskeleton. Moreover, the developed approach provides an idea of three-dimensional arrangement of cellular elements with a greater depth of focus than that of optical methods, which is drastically important for fulfilling the needs of tissue engineering. Being fast and simple, the method is able to change the whole concept of using SEM in biomedical studies.

1.
M.
Shaklai
,
M.
Tavassoli
,
Lanthanum as an electron microscopic stain
.
J. Histochem. Cytochem.
1982
;
30
(
12
):
1325
30
.
2.
C.F.
Doggenweiler
,
S.
Frenk
,
Staining properties of lanthanium on cell membranes
.
PNAS USA
1965
;
53
(
2
):
425
30
.
3.
C.
Zancanaro
,
A.
Sbarbati
,
F.
Franceschini
 et al 
The chemoreceptor surface of the taste disc in the frog, Rana esculenta. An ultrastructural study with lanthanum nitrate
.
Histochem. J.
1990
;
22
(
9
):
480
6
.
4.
R.A.
Leslie
,
The effects of ionic lanthanum and hypertonic physiological salines on the nervous systems of larval and adult stick insects
.
J. Cell Sci.
1975
;
18
(
2
):
271
86
5.
Image acquisition and lanthanoid staining protocols for scanning electron microscopy, available at http://bioree.ru/?lang=en
6.
I.A.
Novikov
,
A.A.
Fedorov
,
A.M.
Subbot
,
I.G.
Griboedova
 Patent RU 2 578 977 C1 (
2015
)
This content is only available via PDF.
You do not currently have access to this content.