title: Semiquantitative analysis of in-situ hybridization results using IMAGE software: a rapid method for counting reduced silver grains over mRNA-positive cells.
abstract: 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.
authors.affiliation: Neuroscience Training Program, Tulane University Medical School, New Orleans, LA 70112.
authors.forename: L R
authors.initials: LR
authors.lastname: Lucas
authors.forename: R R
authors.initials: RR
authors.lastname: Mize
authors.forename: R E
authors.initials: RE
authors.lastname: Harlan
mesh_terms: 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
chemical_list: D012333:RNA, Messenger; D013373:Substance P; D012834:Silver