Possibilities and limitations of the smearing technique for cellular studies in compact tissues: a transmission electron microscopic analysis of liver smears.
During the last decade a sophisticated biophysical technique called fluorescence recovery after photobleaching (FRAP) has become available for the measurement of lateral diffusion constants of lipids and proteins in the cell membrane. The information obtainable by this method is of great importance; however, the applicability of this method is seriously hindered by the fact that it has been elaborated mostly for isolated, individual cells. On the other hand, the use of freshly prepared cells from compact tissues for such studies is highly desirable. Therefore, a special smearing method was devised (using liver cells) for this purpose. The present paper describes (i) the method of standardization of the smearing technique, (ii) the ultrastructural features of the liver cells when studying the liver smears by means of transmission electron microscopy. The possibilities and limitations of this method are discussed. According to the observations, under well-defined conditions the structural integrity of hepatocytes is maintained in the liver smears to such an extent that they are suitable for FRAP experiments.
Zs-Nagy
V
V
First Laboratory of Clinical Physiology, Tokyo Metropolitan Institute of Gerontology, Japan.
Zs-Nagy
I
I
Kitani
K
K
Ohta
M
M
Imahori
K
K
D000818:Animals; D005260:Female; D008099:Liver / Q000166:cytology / Q000648:ultrastructure*; D008297:Male; D008722:Methods; D008854:Microscopy, Electron / Q000379:methods*; D051381:Rats; D011916:Rats, Inbred F344; D011919:Rats, Inbred Strains