Fluorescence microscopy of oligonucleotide probe arrays

Mark O. Trulson; David Stern; Ian D. Walton; Audrey D. Suseno; Richard P. Rava

doi:10.1117/12.273511

7 May 1997 Fluorescence microscopy of oligonucleotide probe arrays

Mark O. Trulson, David Stern, Ian D. Walton, Audrey D. Suseno, Richard P. Rava

Proceedings Volume 2980, Advances in Fluorescence Sensing Technology III; (1997) https://doi.org/10.1117/12.273511
Event: BiOS '97, Part of Photonics West, 1997, San Jose, CA, United States

Abstract

We have been exploiting high density oligonucleotide arrays to carry out sequence analysis of genetic material from diverse sources. The method utilizes the hybridization of fluorophore labelled nucleic acids to the array and interpretation of the resulting spatial pattern of fluorescence. Our ability to obtain sequence information from the array is governed by the interplay of the synthesis and hybridization chemistry, the photophysics of the fluorophores and background interferences, and the performance of the fluorescence imaging system. The high photolithographic resolution and large usable area of the synthesis process and the presence of submonolayer coverages of fluorophores dictate that the fluorescence detection system meet several potentially conflicting performance criteria. High spatial resolution, high sensitivity, large field of view, low chromaticity and image distortion, and high dynamic range are required simultaneously. Suitable nucleic acid-fluorophore conjugates should have high absorption cross sections and emission quantum yields, low photobleaching quantum yields, and resistance to transient saturation under intense illumination. Our approaches to the design and photophysical characterization of the detection process will be discussed within the context of improving the volume of sequence information and detection limits.

Citation Download Citation

Mark O. Trulson, David Stern, Ian D. Walton, Audrey D. Suseno, and Richard P. Rava "Fluorescence microscopy of oligonucleotide probe arrays", Proc. SPIE 2980, Advances in Fluorescence Sensing Technology III, (7 May 1997); https://doi.org/10.1117/12.273511

ACCESS THE FULL ARTICLE

INSTITUTIONAL
Select your institution to access the SPIE Digital Library.

SELECT YOUR INSTITUTION

PERSONAL
Sign in with your SPIE account to access your personal subscriptions or to use specific features such as save to my library, sign up for alerts, save searches, etc.

PERSONAL SIGN IN

No SPIE Account? Create one

PURCHASE THIS CONTENT

SUBSCRIBE TO DIGITAL LIBRARY

50 downloads per 1-year subscription

Members: $195

Non-members: $335 ADD TO CART

25 downloads per 1 - year subscription

Members: $145

Non-members: $250 ADD TO CART

PURCHASE SINGLE ARTICLE

Includes PDF, HTML & Video, when available