Journal of Histochemistry and Cytochemistry Priciples for Free Access to Science
  Search:   
    >> Advanced Search

Guidelines | Subscriptions | About | exPRESS - Current - Archive | Business Information | Contact
This Article
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by MANSBERG, H. P.
Right arrow Articles by KUSNETZ, J.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by MANSBERG, H. P.
Right arrow Articles by KUSNETZ, J.
Social Bookmarking
Add to CiteULike   Add to Complore   Add to Connotea   Add to Del.icio.us   Add to Digg   Add to Reddit   Add to Technorati  
What's this?

QUANTITATIVE FLUORESCENCE MICROSCOPY: FLUORESCENT ANTIBODY AUTOMATIC SCANNING TECHNIQUES

H. P. MANSBERG 1 and J. KUSNETZ 1

1 Airborne instruments Laboratory, Division of Cutler-Hammer, Incorporated, Deer Park, Long island, New York

The application of microscanning techniques to the detection, classification and counting of fluorescence-tagged microorganisms presents unique and somewhat more severe optical and photometric problems than those encountered in absorption type microscanning. Low fluorescence emission, autofluorescence of optics, filters and specimen mounting media and the close proximity of the excitation and emission wavelengths of typical fluorochromes impose severe limitations on sensitivity and maximum scanning rates. By the use of bright field illumination, liquid dye immersion filters, and careful matching of light source and filter spectral characteristics to those of the fluorochromes, fluorescence output was increased 10-100 times with negligible increase in background level. The effectiveness of a number of light sources filters and phototubes in producing maximal signals from fluorescein isothiocyanate-tagged microorganisms in an experimental fluorescence microscanner are compared. Proper selection produced increases in sensitivity of more than 10 times the initial sensitivity obtained with standard fluorescence microscopes and illuminators. Image plane, object plane and flying spot scanning techniques are compared as to their suitability for fluorescence microscanning. Practical scanner designs are illustrated. A consideration of the advantages of laser excitation of fluorochromes indicates that in addition to greatly simplifying filtering problems, a 102 increase in irradiance can be expected. The possibility of differential diagnosis based upon two color fluorescence discrimination is explored. Overlapping emission spectra presents the major difficulties in such a system. An experimental two color fluorescence microscanner and the optical and electronic techniques employed for the discrimination of overlapping color spectra is described.

Submitted on July 6, 1965


Add to CiteULike CiteULike   Add to Complore Complore   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us   Add to Digg Digg   Add to Reddit Reddit   Add to Technorati Technorati    What's this?





Guidelines | Subscriptions | About | exPRESS - Current - Archive | Business Information | Contact
The Journal of Histochemistry & Cytochemistry is owned, published, and licensed by The Histochemical Society © 1966