Rapid continuous monitoring of enzyme activity in tissue sections: experience with the M85A and Zeiss UMSP-30 systems.
We have previously described methods for the continuous monitoring of formazan deposition in tissue sections with a system based on the Vickers M85A microdensitometer. However, this instrument only allows the monitoring of a single field in each section. We have now developed a new system based on the Zeiss UMSP-30 microspectrophotometer. This machine is entirely computer controlled and by virtue of its fast-scanning stage allows the rapid (less than 0.5 s) sequential monitoring of multiple fields (up to 35) in each section. Thus a number of cell types may be studied simultaneously and work which used to take a full working day with the M85A system now can be performed in 45 min. As with the M85A the Zeiss system has full capability for data analysis (i.e. calculation of initial velocity rates, etc.). We have found that continuous monitoring of tissue sections by microdensitometry is a precise, sensitive and biochemically valid method of studying enzyme activity within the cellular matrix.
Lomax
R B
RB
Department of Medicine, University of Manchester, Hope hospital, UK.
Daglish
A
A
Taylor
R J
RJ
Gordon
M T
MT
Robertson
W R
WR
D015096:3-Hydroxysteroid Dehydrogenases / Q000378:metabolism; D000818:Animals; D003720:Densitometry / Q000295:instrumentation / Q000379:methods*; D005260:Female; D006651:Histocytochemistry / Q000379:methods; D015591:Microspectrophotometry / Q000295:instrumentation / Q000379:methods*; D009132:Muscles / Q000166:cytology / Q000201:enzymology; D009246:NADH Tetrazolium Reductase / Q000378:metabolism; D010053:Ovary / Q000166:cytology / Q000201:enzymology; D051381:Rats
D015096:3-Hydroxysteroid Dehydrogenases; D009246:NADH Tetrazolium Reductase