Single-molecule DNA detection in microfabricated capillary electrophoresis chips

Brian B. Haab; Richard A. Mathies

doi:10.1117/12.307316

10 April 1998 Single-molecule DNA detection in microfabricated capillary electrophoresis chips

Brian B. Haab, Richard A. Mathies

Proceedings Volume 3259, Systems and Technologies for Clinical Diagnostics and Drug Discovery; (1998) https://doi.org/10.1117/12.307316
Event: BiOS '98 International Biomedical Optics Symposium, 1998, San Jose, CA, United States

Abstract

Single-molecule fluorescence burst counting is a highly sensitive method for detecting electrophoretic separations of ds-DNA fragments. In previous work we demonstrated the detection of single DNA fragments as small as 100 bp in capillary electrophoresis separations. To further enhance the applicability of this method to low level pathogen and mutation detection, we have now successfully performed single molecule detection of DNA separations in microfabricated glass capillary electrophoresis (CE) chips. By fabricating CE chips with a 200 micrometers thick top cover plate and by using a 40X, 1.3 NA immersion microscope objective, the S/N ratio for single molecule detection is enhanced by more than two-fold over conventional capillaries. By constricting the sample in the detection region to an approximately 10 micrometers wide by approximately 10 micrometers deep cross section, approximately 10% of the molecules passing through the channel are probed by the approximately 2 micrometers wide focused laser beam. This is a 1000-fold improvement over our previous work. We have now achieved an on-column detection limit of approximately 600 fM or 500 molecules for 500 bp DNA fragments (S/N equals 3).

Citation Download Citation

Brian B. Haab and Richard A. Mathies "Single-molecule DNA detection in microfabricated capillary electrophoresis chips", Proc. SPIE 3259, Systems and Technologies for Clinical Diagnostics and Drug Discovery, (10 April 1998); https://doi.org/10.1117/12.307316

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