Novel method for quantification of brain cell swelling in rat hippocampal slices.
We have developed a novel device for the quantification of edematous morphology changes in acute brain slices. We can also carry out real-time monitoring of detailed hippocampal cells. The device we developed is based on infrared differential interference contrast microscopy (IR-DIC) and a custom-made real-time computerized image-analysis system for quantification of the morphological dynamics of cells in slice preparations. We applied the coefficient of variation (CV) of light intensity in IR-DIC images to evaluate the change in morphological dynamics. We examined three kinds of edema in the CA1 region of rat hippocampal acute slices under conditions of hypotonic, strong excitation, and experimental ischemia, together with field excitatory postsynaptic potential (fEPSP) recording from radiatum in CA1 the region. There were notable close relationships among the edema formations, the light transmittance, the extent of changes in CV, and features of fEPSP during the three different insults. The present results indicate that CV is a reliable quantification index for edema formation in brain tissue and confirm that applying CV for the analysis in addition to the light transmittance analysis presents additional important information on brain tissue swelling.
Nakajima
Ryuichi
R
Laboratory of Cellular Neurobiology, School of Life Science, Tokyo University of Pharmacy and Life Science, Hachioji-shi, Tokyo, Japan.
Nakamura
Takeshi
T
Ogawa
Miyuki
M
Miyakawa
Hiroyoshi
H
Kudo
Yoshihisa
Y
D000818:Animals; D004487:Edema / Q000209:etiology / Q000473:pathology*; D019706:Excitatory Postsynaptic Potentials / Q000187:drug effects; D006624:Hippocampus / Q000473:pathology* / Q000503:physiopathology; D006982:Hypertonic Solutions / Q000009:adverse effects; D007091:Image Processing, Computer-Assisted; D066298:In Vitro Techniques; D007511:Ischemia / Q000473:pathology / Q000503:physiopathology; D008297:Male; D008857:Microscopy, Interference / Q000379:methods*; D009474:Neurons / Q000187:drug effects / Q000473:pathology* / Q000502:physiology; D011188:Potassium / Q000494:pharmacology; D051381:Rats; D017208:Rats, Wistar; D013997:Time Factors; D014741:Video Recording / Q000295:instrumentation / Q000379:methods
D006982:Hypertonic Solutions; D011188:Potassium