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Proceedings Paper

Imaging leukocytes in vivo with third harmonic generation microscopy
Author(s): 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
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Paper 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.

Paper Details

Date Published: 22 February 2013
PDF: 7 pages
Proc. SPIE 8588, Multiphoton Microscopy in the Biomedical Sciences XIII, 85881R (22 February 2013); doi: 10.1117/12.2003809
Show Author Affiliations
Cheng-Kun Tsai, National Taiwan Univ. (Taiwan)
Chien-Kuo Chen, National Taiwan Univ. (Taiwan)
Yu-Shing Chen, National Taiwan Univ. (Taiwan)
Pei-Chun Wu, National Taiwan Univ. (Taiwan)
Tsung-Yuan Hsieh, National Taiwan Univ. (Taiwan)
Han-Wen Liu, National Taiwan Univ. (Taiwan)
Chiou-Yueh Yeh, National Taiwan Univ. (Taiwan)
Win-Li Lin, National Taiwan Univ. (Taiwan)
Jean-San Chia, National Taiwan Univ. (Taiwan)
Tzu-Ming Liu, National Taiwan Univ. (Taiwan)

Published in SPIE Proceedings Vol. 8588:
Multiphoton Microscopy in the Biomedical Sciences XIII
Ammasi Periasamy; Karsten König; Peter T. C. So, Editor(s)

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