Imaging leukocytes in vivo with third harmonic generation microscopy

Cheng-Kun Tsai; Chien-Kuo Chen; Yu-Shing Chen; Pei-Chun Wu; Tsung-Yuan Hsieh; Han-Wen Liu; Chiou-Yueh Yeh; Win-Li Lin; Jean-San Chia; Tzu-Ming Liu

doi:10.1117/12.2003809

22 February 2013 Imaging leukocytes in vivo with third harmonic generation microscopy

Cheng-Kun Tsai, Chien-Kuo Chen, Yu-Shing Chen, Pei-Chun Wu, Tsung-Yuan Hsieh, Han-Wen Liu, Chiou-Yueh Yeh, Win-Li Lin, Jean-San Chia, Tzu-Ming Liu

Author Affiliations +

Proceedings Volume 8588, Multiphoton Microscopy in the Biomedical Sciences XIII; 85881R (2013) https://doi.org/10.1117/12.2003809
Event: SPIE BiOS, 2013, San Francisco, California, United States

Abstract

Without a labeling, we demonstrated that lipid granules in leukocytes have distinctive third harmonic generation (THG) contrast. Excited by a 1230nm femtosecond laser, THG signals were generated at a significantly higher level in neutrophils than other mononuclear cells, whereas signals in agranular lymphocytes were one order smaller. These characteristic THG features can also be observed in vivo to trace the newly recruited leukocytes following lipopolysaccharide (LPS) challenge. Furthermore, using video-rate THG microscopy, we also captured images of blood cells in human capillaries. Quite different from red-blood-cells, every now and then, round and granule rich blood cells with strong THG contrast appeared in circulation. The corresponding volume densities in blood, evaluated from their frequencies of appearance and the velocity of circulation, fall within the physiological range of human white blood cell counts. These results suggested that labeling-free THG imaging may provide timely tracing of leukocyte movement and hematology inspection without disturbing the normal cellular or physiological status.

Citation Download Citation

Cheng-Kun Tsai, Chien-Kuo Chen, Yu-Shing Chen, Pei-Chun Wu, Tsung-Yuan Hsieh, Han-Wen Liu, Chiou-Yueh Yeh, Win-Li Lin, Jean-San Chia, and Tzu-Ming Liu "Imaging leukocytes in vivo with third harmonic generation microscopy", Proc. SPIE 8588, Multiphoton Microscopy in the Biomedical Sciences XIII, 85881R (22 February 2013); https://doi.org/10.1117/12.2003809

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

Members: $17.00

Non-members: $21.00 ADD TO CART

PROCEEDINGS
7 PAGES

DOWNLOAD PAPER SAVE TO MY LIBRARY

GET CITATION

RIGHTS & PERMISSIONS

Get copyright permission Get copyright permission on Copyright Marketplace

KEYWORDS

Blood

In vivo imaging

Microscopy

Harmonic generation

Capillaries

Confocal microscopy

Luminescence

Show All Keywords

Keywords/Phrases

Search In:

Publication Years