Semiquantitative analysis of in-situ hybridization results using IMAGE software: a rapid method for counting reduced silver grains over mRNA-positive cells.
The advent of microcomputers has brought about a revolution in the computing power available to the average user. Image analysis is a very resource-intensive process, making great demands on computing power, memory, and display capabilities of most computers. Thus, in the past, dedicated, single-use hardware and software had to be custom made for environments requiring image analysis. We present here an easy-to-use image analysis protocol available to most users with a Macintosh II series computer and access to IMAGE (a public domain image analysis program). The protocol allows for semi-quantitation of silver grains over cells used in the interpretation of in-situ hybridization results. We show that the method provides a quick and reliable means of counting grains over mRNA-positive cells in an automated fashion. We also provide evidence that the method can be used to detect differences between experimental treatments.
Lucas
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Neuroscience Training Program, Tulane University Medical School, New Orleans, LA 70112.
Mize
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Harlan
R E
RE
D000818:Animals; D002421:Caudate Nucleus / Q000378:metabolism; D005069:Evaluation Studies as Topic; D007091:Image Processing, Computer-Assisted / Q000295:instrumentation* / Q000379:methods* / Q000592:standards; D017403:In Situ Hybridization / Q000379:methods*; D008297:Male; D011699:Putamen / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D012834:Silver; D012984:Software; D013373:Substance P / Q000235:genetics
D012333:RNA, Messenger; D013373:Substance P; D012834:Silver