Methylene Blue-stained Interstitial Cells are Electrically Active in the Myenteric Board Freshly Prepared from the Murine Small Intestine

Many gastrointestinal muscles show electrical oscillation, so-called "slow wave", originated from interstitial cells of Cajal (ICCs). Thus, a technique to freshly isolate the cells is indispensable to explore the electrophysiological properties of the ICCs. To apply an enzyme solution on the serosal surface for cell isolation, the intestine was inverted and 0.02% trypsin solution and 0.04% collagenase solution were applied to serosal cavity. After the enzyme treatment, mucosal layer was removed and longitudinal muscle layer was gently separated from the rest of tissue. The thin layer was stretched in the recording chamber and mounted on an inverted microscope. Using beta-escine, perforated whole cell patch clamp technique was used. Under a microscope, the tissue showed smooth muscle cells and interstitial cells around the myenteric plexus. Under voltage clamp condition, three types of membrane potential were recorded. One group of interstitial cells, which were positive to methylene blue and CD34, showed spontaneous outward current. These cells had bipolar shape and were considered as fibroblast-like cells because of their peculiar shape and arrangement. Another group, positive to c-kit and methylene blue, showed spontaneous inward current. These cells had more rounded shape and processes and were considered as ICCs. The third, positive to c-kit and had granules containing methylene blue, showed quiet membrane potentials under the voltage-clamp mode. These cells appeared to be resident macrophages. Therefore, in the freshly isolated thin tissue preparation, methylene blue could easily identify three types of cells rather than morphological properties. Using this method, we were able to study electrical properties of fibroblast and residential macrophage as well as myenteric ICCs.

Biomicroscopic Examination of Precipitates on the intraocular Lens

We studied the foregin body reaction to the implanted intraocular lens(IOL) and IOL acceptance in the patient's eye by biomicroscopic examination of precipitates on the IOL surface in 432 cases. We divided the cellular and pigment precipitates of the posterior chamber lens(PCL) showed grade 3 until seven days postoperative and then slowly decreased; but, that of the anterior chamber lens(ACL) persisted a little longer. Pigment precipitates on ACL was less than that of PCL, and pigment precipitates stayed longer than cellular precipitates. Round, bipolar small cellular precipitates seem to be identical to fibroblast-like cells, and large, irregular shaped cellular precipitates seem to be foreign body giant cells. A large number of foreign body giant cells precipitated and persisted longer in cases of prolonged postoperative inflammation.

Biomicroscopic Examination of Precipitates on the intraocular Lens

We studied the foregin body reaction to the implanted intraocular lens(IOL) and IOL acceptance in the patient's eye by biomicroscopic examination of precipitates on the IOL surface in 432 cases. We divided the cellular and pigment precipitates of the posterior chamber lens(PCL) showed grade 3 until seven days postoperative and then slowly decreased; but, that of the anterior chamber lens(ACL) persisted a little longer. Pigment precipitates on ACL was less than that of PCL, and pigment precipitates stayed longer than cellular precipitates. Round, bipolar small cellular precipitates seem to be identical to fibroblast-like cells, and large, irregular shaped cellular precipitates seem to be foreign body giant cells. A large number of foreign body giant cells precipitated and persisted longer in cases of prolonged postoperative inflammation.