Counting cells in sectioned material: a suite of techniques, tools, and tips.
This unit presents protocols to obtain accurate estimates of cell density and cell number in sectioned material by using a light microscope. The "optical disector" or "3-D counting method" is described, followed by Abercrombie's less commonly used two-section comparison (TSC) method. These basic protocols are accompanied by four support protocols: one for celloidin embedding, which renders superb morphology, one for point counting, which is important for volume measurements and is almost always used in conjunction with the disector or 3-D counting, one for handling the potential problem of z-axis distortion and the consequences that this error can have on density estimates and sampling tactics when using the disector, and finally, one that provides a guide for calibrating and verifying estimates obtained by counting methods.
Williams
Robert W
RW
University of Tennessee Health Science Center, Memphis, Tennessee, USA.
von Bartheld
Christopher S
CS
Rosen
Glenn D
GD
D000818:Animals; D048588:Benzoxazines; D035843:Biomedical Research / Q000379:methods*; D001921:Brain / Q000166:cytology; D002138:Calibration; D002452:Cell Count / Q000295:instrumentation* / Q000379:methods*; D002490:Central Nervous System / Q000166:cytology*; D003101:Collodion; D004396:Coloring Agents; D006801:Humans; D021621:Imaging, Three-Dimensional; D042282:Microdissection / Q000295:instrumentation / Q000379:methods; D009445:Neuroanatomy / Q000379:methods*; D010078:Oxazines; D013194:Staining and Labeling; D016610:Tissue Embedding / Q000379:methods
D048588:Benzoxazines; D004396:Coloring Agents; D010078:Oxazines; C028911:cresyl violet; D003101:Collodion